137 results on '"Schulz LC"'
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2. Versuche zur Erzeugung von chronischem Rotlauf beim Schwein
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G. Trautwein, Dranke D, Schulz Lc, F. Weiland, C. Messow, and K. H. Böhm
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Philosophy ,medicine ,medicine.disease ,Erysipelas ,Humanities ,Antibody formation - Abstract
Zusammenfassung In drei Versuchen mit 39 serologisch negativen SPF-Schweinen (SPF-Primartiere) wurde durch wiederholte iv. und sc. oder intraartikulare Verabreichung von Rotlauf-Totantigen versucht, Veranderungen wie bei chronischem Rotlauf zu erzeugen. Einer der drei Versuche wurde in Anlehnung an einen von Freeman und Mitarbeiter (1964 a) durchgefuhrten Versuch konzipiert. Die Versuchsergebnisse liefern keine Anhaltspunkte fur eine Mitbeteiligung der Allergie. Die von Freeman und Mitarbeiter (1964 a) aus ihren Versuchsergebnissen gezogenen Schlusfolgerungen werden in Ubereinstimmung mit Wellmann und Mitarbeiter (1965) abgelehnt. Auf Grund der in der Synovialmembran von Schweinen mit chronischer Rotlaufpolyarthritis nachweisbaren Plasmazellansammlungen wird es in Ubereinstimmung mit Timoney (1967) fur wahrscheinlich gehalten, das die lokale Antikorperbildung eine das Krankheitsbild mitgestaltende Rolle spielt. Summary Studies on the induction of chronic erysipelas in pigs I. Experiments with killed antigen In three experiments with 39 serologically negative SPF pigs (primary SPF animals) killed erysipelas antigen was inoculated i. v., s. c. or intraarticularly in order to see whether the changes typical of chronic erysipelas could be produced. One of the three attempts was based on the experiment conducted by Freeman and Berman (1964 a). The results give no support to view that allergy played a part. Like Wellmann et. al (1965), we could not accept the conclusions which Freeman and Berman (1964 a) drew from their own experiments. On the basis of the collections of plasma cells found in the synovial membrane of pigs with chronic erysipelas polyarthritis, it is concluded, in agreement with Timoney (1967) that local antibody formation probably plays a part in the disease picture. Resume Essai d'induction de rouget chronique — chez le pore I. Essai avec un antigene tue On a essaye de provoquer des lesions de rouget chronique au cours de trois essais avec 39 porcs SPF (animaux primaires) serologiquement negatifs par des applications i/v, s/c ou intraarticulaires d'antigene tue. Un des trois essais a ete concu comme appui d'une experience de Freeman et collab. (1964 a). Les resultats ne presenterent aucune intervention sous forme d'allergie. Les conclusions tirees par Freeman et collab. (1964 a) de leurs resultats d'experience ne concordent pas avec celles de Wellmann et collab. (1965). Sur la base des assemblages de cellules du plasma dans la membrane synoviale des procs atteints de polyarthrite chronique due a un rouget, on estime comme vraisemblable, en accord avec Timoney (1967), que la formation locale d'anticorps joue un role dans l'image de la maladie. Resumen Ensayos para inducir mal rojo cronico en el cerdo I. Tentativas con antigenos muertos En tres ensayos con 39 cerdos gnotobioticos (animales SPF primarios) serologicamente negativos, se intento producir modificaciones como las que se registran en el mal rojo cronico mediante aplicaciones repetidas por las vias intravenosa y subcutanea o intraarticular de antigeno muerto de mal rojo. Uno de los tres ensayos se concibio inspirandose en otro ejecutado por Freeman y colaboradores (1964 a). Los resultados experimentales no ofrecen ningun punto de engarce que pruebe la coparticipacion de la alergia. En concordancia con Wellmann y colaboradores (1965), se refutan las conclusiones que establecieron a raiz de los resultados obtenidos en sus experiencias. En vista de las acumulaciones demostrables de celulas plasmaticas en la membrana sinovial de cerdos con poliartritis cronica de mal rojo se cree probable, coincidiendo con Timoney (1967), que la formacion local de anticuerpos juega un papel matizante en el cuadro nosologico.
- Published
- 2010
3. Elektronenmikroskopische Untersuchungen zur endometrialen zytoplastischen Potenz beim Rind*)
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Schulz Lc
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General Veterinary ,Chemistry ,De regulation ,Lymph ,Molecular biology - Abstract
Zusammenfassung 1 Die Funktion endometrialer Stromazcllen und -gefase wird auf Grund elektronenmikroskopischer und zytometrischer Untersuchungen an 72 Rindern in 3 Phasen eingeteilt: a. Fruhe Aufbauphase (amitotische Zellvermehrung); b. Spate Aufbauphase (Leistungswachstum durch Zellvergroserung und Differenzierung); c. Umbauphase (Regression und funktionelle Ruhe). 2 Das retikulare Stroma ist kein synzytialer Zellverband. Es kann nach dem Sinusoidcharakter seiner Gefase und der von ihnen ausgehenden intravasalen sowie adventitiellen Zytogenese dem retikulohistiozytaren System im engeren Sinne zugeordnet werden. 3 In phagozytierenden Histiozyten wird die zyklusabhangige Ausreifung des Ceroidpigmentes verfolgt und die Bedeutung der Mitochondrien bei der Speicherung von Eisenmizellen besprochen. 4 Die lymphozytare und plasmazellulare Histiogenese last sich im gesunden Endometrium und innerhalb von Lymphknotchen nachweisen. Ein Auftreten von Lymphknotchen wird als Anzeichen gestorter Regulation angesprochen. 5 Mastzellenausbildung erfolgt in deutlicher Abhangigkeit vom Zyklus. Der Degranulierung im Hohepunkt der Progesteronphase geht ein histochemisch nachweisbarer Ausreifungsprozes voraus. 6 Die extramedullare Genese eosinophiler Leukozyten kann im Endometrium als sicher gelten. Elektronenmikroskopisch lassen sich ihre adventitielle Entwicklung und Granulaausreifung sowie eine noch ungeklarte Beziehung zum Blutabbau beobachten. Summary Electron microscopical studies on the cytoplastic potency of the endometrium in the cow 1 The function of the endometrial stromal cells and blood vessels is divided into three phases as a result of electron microscopical and cytometric studies on 72 cattle. a. Early constructive phase (amitotic proliferation). b. Late constructive phase (functional growth by increase in cell volume and differentiation). c. Transformation phase (regression and functional rest). 2 The reticular stroma is not a syncytium. The sinusoidal character of its vessels and the intravascular and adventitial changes that develop from them point to a relationship to the reticulo-histiocytic system. 3 In histiocytes capable of phagocytosis it is possible to follow the maturation of ceroid pigment in the course of the sexual cycle and to show the importance of mitochondria in the storage of iron-containing micelles. 4 Lymphocytic and plasma cell histogenesis can be seen in the healthy endometrium and in the lymph nodes. The appearance of lymphoid follicles is interpreted as an indication of disordered regulation. 5 The formation of mast cells is very closely dependant on the cycle. The loss of granules at the height of the progesterone phase is by a process maturation which can be detected histochemically. 6 The extra-medullary formation of eosinophile leucocytes in the endometrium is a certainty. Under the electron microscope one can observe their development and the maturation of their granules, but their relation to the destruction of red blood cells is still obscure. Resume Recherches au microscope electronique sur la potentialite cytoplastique de l'endometre chez le boeuf 1 La fonction des cellules et des vaisseaux du stroma de l'endometre a ete divisee en trois phases sur la base de recherches au microscope electronique et d'etudes cytometriques effectuees sur un total de 72 bovins. a. Phase constructive precoce (multiplication cellulaire amitotique); b. Phase constructive tardive (Croissance fonctionnelle par augmentation du volume cellulaire et differenciation); c. Phase de transformation (regression et repos fonctionel). 2 Le stroma reticulaire n'est pas un syncytium. Le caractere sinusoide de ses vaisseaux, la cytogenese intravasculaire et adventitielle qu'on peut y constater permettent de l'assimiler au systeme reticulo-histiocytaire proprement dit. 3 Dans les histiocytes capables de phagocytose on peut suivre la maturation du pigment ceroiee en rapport avec le cycle sexuel et l'importance des mitochondries dans le stockage de micelles sideroferes est discutee. 4 L'histogenese lymphocytaire et plasmacellulaire peut etre mise en evidence dans l'endometre sain et dans les follicules lymphatiques. L'apparition de follicules lymphatiques est interpretee comme un signe de regulation perturbee. 5 La formation des mastocytes est en relation tres nette avec le cycle. La degranulation qui se produit a l'apogee de la phase du progesterone est precedee d'un processus de maturation decelable par les methodes histo-chimiques. 6 La genese extramedullaire des leucocytes eosinophiles peut etre consideree comme certaine dans l'endometre. Au microscope electronique on peut observer leur devoloppement adventitiel et la maturation des grains, ainsi qu'une relation encore obscure avec la destruction des globules sanguins. Resumen Estudios con el microscopio electronico sobre la potencia citoplastica endometrial en la vaca 1 La funcion de las celulas y vasos del estroma endometrial se divide, con arreglo a las investigaciones microscopico-electronicas y citometricas en 72 vacas, en 3 fases: a. fase precoz de estructuracion (aumento celular amitotico); b. fase tardia de estructuracion (crecimiento del rendimiento del rendimiento mediante aumento del tamano celular y diferenciacion); c. fase de transformacion (regresion y reposo funcional). 2 El estroma reticular no es ninguna asociacion celular sincitial. Segan el caracter sinusoide de sus vasos y la citogenesis intravasal y adventicia procedente de ellos, se puede adscribir, en sentido estricto, al sistema reticulohistiocitario. 3 En histiocitos fagoeitantes se persigue la maduracion del pigmento ceroide dependiente del ciclo y se discute la importancia de las mitocondrias en el almacenamiento de las micelas ferreas. 4 La histiogenesis linfocitaria y plasmacelular se puede identificar en el endometrio sano y dentro de los nodulos linfaticos. La aparicion de nodulos linfaticos se interpreta como un signo de regulacion alterada. 5 La formacion de celulas cebadas se efectaa en marcada dependencia del ciclo. A la degranulacion en el culmen de la fase luteinica le precede un proceso de maduracion identificable por via histoquimica. 6 Se puede admitir como segura la genesis extramedular de los leucocitos eosinofilos en el endometrio. Con el microscopio electronico se puede observar su desarrollo adventicio y la maduracion de los granulos, asi como una interrelacion no aclarada con el catabolismo sanguineo.
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- 2010
4. Schocksyndrom und seine pathogenetische Bedeutung bei Poliomyelomalazie und anderen Erkrankungen des Schweines
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Schulz Lc and Heinrich Behrens
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Poliomyelomalacia ,Gynecology ,Dorsum ,medicine.medical_specialty ,General Veterinary ,Philosophy ,medicine ,Sindrome de - Abstract
Zusammenfassung Die Bedeutung des Schocksyndromes wird bei der Poliomyelomalazie und anderen Erkrankungen des Schweines, insbesondere beim sog. plotzlichen Herztod des Schweines besprochen. Kennzeichnend fur das Schocksyndrom ist der pathologisch-anatomische Befund im zentralen Nervensystem, die Gewebseosinophilie und die Hyperaemie im Splanchnikusgebiet mit Austritt von Flussigkeit oder Blut in das Darmlumen. Es wird versucht, am Beispiel der Poliomyelomalazie diese Symptome pathogenetisch einzuordnen. In den Anfangsstadien der Poliomyelomalazie lassen sich durch Schock bedingte Storungen der Gefaswandpermeabilitat besonders in der grauen Ruckenmarkssubstanz nachweisen. Sterben die Tiere nicht bei Krankheitsbeginn, so folgen den Permeabilitatsstorungen Umbauprozesse an den Gefaswanden. Diese werden den sog. Collagen-diseases zugerechnet und als rheumatoides Substrat im Sinne der Adaptationskrankheiten nach Selye aufgefask. Nach langerem Krankheitsverlauf konnen Anteile der grauen Ruckenmarkssubstanz infolge anhaltender Durchblutungsstorungen vollstandig erweichen. Aetiologische Beziehungen zur Serosengelenksentzundung werden erwogen. Summary The shock syndrome and its pathogenetic importance in poliomyelomalacia and other diseases of the pig The importance of this syndrome is discussed in the pig, with special reference to the so-called “sudden heart failure”. Characteristics of this syndrome are the pathological anatomical finding in the central nervous system, the tissue eosinophilia and the hyperaemia in the splanchnic region with outpouring of fluid or blood into the lumen of the gut. The authors attempt to explain the pathogenesis of these symptoms, using the example of poliomyelomalacia. In the early stages of poliomyelomalacia disturbances of vascular permeability can be demonstrated, especially in the grey matter of the spinal cord. If the animals do not die in the early stages of the disease, disturbances of permeability are followed by structural changes in the vascular walls. These later are grouped with the so-called “collagen diseases” and are seen as a rheumatoid syndrome in the sense of Selye's adaptation diseases. If the course of the disease is prolonged, parts of the grey matter of the spinal cord may show a complete softening as a result of circulatory disturbances. Aetiological relations to inflammation of serous membranes and joints are discussed. Resume Le syndrome de choc et son importance pathogenique dans la poliomyelomalacie et autres maladies du porc Les auteurs discutent l'importance du syndrome de choc dans la poliomyelomalacie et autres maladies du porc, en particulier la mort cardiaque subite (Herztod). Les signes caracteristiques du syndrome de choc sont les lesions anatomo-pathologiques du systeme nerveux central, l'eosinophilie tissulaire et l'hyperemie des organes places sous la dependance du nerf splanchnique, accompagnees d'une exsudation sereuse ou hemorragique dans la lumiere intestinale. Les auteurs essaient de donner le deroulement pathogenique de ces symptomes dans l'exemple de la poliomyelomalacie. Dans les stades initiaux de la poliomyelomalacie on constate des troubles de la permeabilite vasculaire dus aux chocs et localises surtout dans la substance grise de la moelle epiniere. Si les animaux ne succombent pas au debut de la maladie, les troubles de la permeabilite sont suivis de processus de transformation dans les parois vasculaires. Ceux-ci sont assimiles aux “maladies du collagene” et consideres comme etant de nature rhumatoide dans le sens de la maladie de l'adaptation selon Selye. Apres une maladie assez longue, la substance grise de la moelle epiniere peu subir par endroit, un ramollissement complet par suite de troubles constants de la circulation. Des rapports etiologiques eventuels avec l'inflammation des sereuses et des articulations sont l'objet d'une etude critique. Resumen Sindrome de choque y su significado en la poliomielomalazia y ostras enfermedades porcinas Se discute la significacion del sindrome de choque en la poliomielomalazia y otras enfermedades de los suinos, sobre todo en la llamada muerte cardiaca repentina del cerdo. Caracteristico del sindrome de shock es el hallazgo anatomopatologico en el sistema nervioso central, la eosinofilia histica y la hiperemia en la region esplacnica con salida de liquido o sangre a la luz intestinal. En el ejemplo de la polimielomalazia se intenta encuadrar patogenicamente estos sintomas. En los estadios iniciales de la polimielomalazia, las alteraciones de la permeabilidad de la pared vascular condicionadas por choque se pueden identificar sobre todo en la substancia gris de la medula dorsal. Si los animales no mueren al comienzo de la enfermedad, a las alteraciones de la permeabilidad le siguen procesos de reconstitucion en las paredes vasculares. Estos se adscriben a las denominadas enfermedades colagenas y se conceptuan como substrato reumatoide en el sentido de las enfermedades de adaptacion segun Selye. Tras una patocronia duradera pueden reblandecerse por completo partes de la substancia gris de la medula espinal como consecuencia de alteraciones continuas del riego sanguineo. Se aducen relaciones etiologicas con la artritis serosa.
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- 2010
5. Über die Induktion einer chronischen Polyarthritis mit Bestandteilen von Rotlaufbakterien (Erysipelothrix rhusiopathiae)
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G. Kerlen, Schulz Lc, K. H. Böhm, Jessen H, and W. Hermanns
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biology ,business.industry ,Arthritis ,Erysipelothrix rhusiopathiae ,medicine.disease ,Body weight ,biology.organism_classification ,Molecular biology ,Erysipelas ,Subacute arthritis ,High dosage ,medicine ,Polyarthritis ,business ,Rheumatoide arthritis - Abstract
Zusammenfassung Ein Ultraschall-Gesamtaufschlus von Rotlaufbakterien (Stamm T 28) ist in der Lage, bei Ratten eine subakute Arthritis von rheumatoidem Charakter auszulosen. Vorbedingung ist erstens eine ausreichend hohe Dosierung und zweitens mus eine bestimmte Anflutungshohe des Materials im Organismus gewahrleistet sein. Klinische Erscheinungen, auser einer vorubergehenden Korpergewichtsreduktion, sind selten. Das Antikorperbildungsvermogen entspricht in der akuten Phase dem von infizierten Ratten, am Ende des Versuchszeitraumes (56 Tage nach der letzten Inokulation) sinkt der Titer auf Normal werte ab. Histopathologisch sind nur die Gelenke verandert. Es finden sich einmal akute, exsudative Prozesse mit Fibrinabscheidung, Neutrophilenemigration und Deckzelldesquamation; zum anderen Veranderungen, die fur eine rheumatoide Arthritis charakteristisch sind, wie Deckzellhypertrophie und -hyperplasie, lymphoplasmazellulare Zellinfiltrate im subsynovialen Bindegewebe und Pannusbildung. Weitergehende, destruktive Gelenkveranderungen wurden nicht beobachtet. Aufgrund des Applikationsmusters wird die Aktivierung von Systemen diskutiert, die eine Ablagerung des Erregermaterials im Gelenk begunstigen. Die weiteren Charakteristika dieser Arthritis deuten auf eine immunpathologische Reaktion vom Typ III (Arthus-Reaktion) hin. Summary Induction of chronic polyarthritis with constituents of erysipelas bacteria (Erysipelothrix rhusiopathiae) 2. Studies on induction of arthritis in rats An ultrasonicated preparation of erysipelas bacteria (strain T 28) can produce a subacute arthritis of rheumatoid character in rats. The first requirement is a sufficiently high dosage and the second is a sufficient level of diffusion of the material within the animal. Clinical signs are rare, apart from a temporary loss of body weight. The ability to produce antibody in the acute phase is similar to that in infected rats; at the end of the 56 days experimental period the titre had fallen to normal. Histopathological changes are confined to the joints. On the one hand there are acute, exudative changes, with fibrin deposition, neutrophil emigration and lining cell desquamation; other changes, characteristic of rheumatoid arthritis, such as lining cell hypertrophy and hyperplasia, lympho-plasma cell infiltration into the sub-synovial connective tissue and pannus formation, were also seen. Extensive destructive changes in the joints were not found. On the basis of the technique of administration, the use of systems which favour widespread distribution of the causative material into the joints is discussed. The further characteristics of this arthritis suggest an immunopathological reaction of Type III (Arthus reaction). Resume A propos de l'induction d'une polyarthrite chronique avec des elements de bacteries du rouget (Erysipelothrix rhusiopathiae) Communication II: Recherches sur l'induction de l'arthrite chez des rats Une decomposition totale de bacteries du rouget (souche T 28) par des ultrasons est en mesure de declencher une arthrite subaigue a caractere rhumatoide chez des rats. La condition est tout d'abord une dose suffisamment elevee et ensuite un taux determine de diffusion du materiel dans l'organisme. Les signes cliniques, mis a part un amaigrissement momentane, sont rares. La capacite de formation des anticorps correspond dans la phase aigue a celle des rats infectes, le titre s'abaissant aux valeurs normales a la fin de la periode d'essai (56 jours apres la derniere inoculation). Seules les articulations sont modifiees histopathologiquement. On a constate une fois des processus aigus, exsudatifs avec formation de fibrine, migration de neutrophiles et desquamation des cellules de couverture; d'autres lesions caracteristiques d'une arthrite rhumatoide ont ete une hypertrophie et une hyperplasie des cellules de couverture, une infiltration de cellules lymphoplasmiques dans le tissu conjonctif subsynovial et une formation d'un pannus. D'autres lesions destructives articulaires n'ont pas ete observees. On discute sur la base de l'echantillon d'application l'activation des systemes qui favorisent un depot du materiel bacterien dans une articulation. Les autres caracteres de cette arthrite font penser a une reaction immunopathologique du type III (reaction d'Arthus). Resumen Sobre la induccion de una poliartritis cronica con los componentes de bacterias mal rojo (Erysipelothrix rhusiopathiae) Comunicacion 2°: Ensayos sobre la induccion de artritis en ratas Una descomposicion total por ultrasonido de las bacterias mal rojo (estirpe T 28) esta en condiciones de desencadenar en las ratas una artritis subaguda de caracter reumatoide. Condiciones previas son primero el que este garantizada una dosificacion suficientemente elevada y segundo una tasa determinada de difusion del material en el organismo. Son raros los signos clinicos, excepto la reduccion pasajera del peso corporal. La capacidad de anticuerpogenesis corresponde en la fase aguda a la de las ratas infectadas; al final del espacio de tiempo que dura es ensayo (56 dias contando desde la inoculacion ultima) vuelve a descender el titulo a los valores normales. Histopatologicamente solo se hallan modificadas las articulaciones. Por un lado se constatan procesos exudativos agudos con formacion de fibrina, migracion de neutrofilos y descamacion de las celulas de revestimiento; por otro lado, modificaciones que son caracteristicas de una artritis reumatoide, tales como la hipertrofia e hiperplasia de las celulas de revestimiento, infiltrados celulares linfoplasmaticcs en el tejido conjuntivo subsinovial v formacion de un pannus. No se observaron otras modificaciones articulares destructivas continuativas. A la vista del modelo de aplicacion, se discute la activacion de los sistemas que favorecen el disposito del material bacteriano en la articulacion. Las caracteristicas restantes de esta artritis senalan hacia una reaccion inmunopatologica del tipo III (reaccion de ARTHUS).
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- 2010
6. Vergleichende Betrachtungen zum Sarkombefall bei Mensch und Hund
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Schulz Lc
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Gynecology ,Cancer Research ,medicine.medical_specialty ,Oncology ,Philosophy ,medicine ,General Medicine - Abstract
Im ersten Teil wird der seit 1948 auserordentlich starke Anstieg der Zahl mit Geschwulsten behafteter Hunde im Sektionsgut diskutiert. Als Ursache ist die sich jahrlich verkleinernde Anzahl der im jugendlichen Alter an Staupe verstorbenen uud sezierten Tiere und somit eine Erhohung des durchschnittlichen Lebensalters des Sektionsgutes anzusehen. Dabei kann die Erwartung der vergleichenden Onkologie nicht bestatigt werden, das sich mit hoherer Lebenserwartung beim Hund analog zum Menschen die prozentualen Anteile der Sarkome auf Kosten der Carcinome verringern. Wenn diese an unserem Material erhobene Tatsache auch von anderen veterinarpathologischen Instituten bestatigt werden kann, muste man annehmen, das die Neigung des alternden Menschen zu epithelialen Blastomen fur den Hund nicht zutrifft.
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- 1958
7. Fluorescence-activated nuclear sorting (FANS) of nuclei from in vitro-generated syncytiotrophoblast.
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Khan T, Whyte JJ, Schulz LC, and Roberts RM
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Large, multinucleated cells, like syncytiotrophoblasts (STB), are not readily analyzed by standard methods used for single cells, such as single-cell RNA-sequencing and fluorescence-activated cellular sorting (FACS). Here we have demonstrated that fluorescence-activated nuclear sorting (FANS) is suitable to analyze nuclei from STB. Human pluripotent stem cells (PSCs) can be differentiated into a mixed trophoblast populations comprising approximately 20 % STB by treatment with BMP4 (Bone Morphogenetic Protein-4), plus A83-01 and PD173074, inhibitors of activin and FGF2 signaling, respectively (the BAP model) in about a week. Here we demonstrate that FANS can be used to separate two types of STB nuclei from the nine different clusters of trophoblast nuclei previously identified in the BAP model by single nucleus RNA sequencing (snRNAseq). Rather than using cell surface markers, as in FACS, transcription factors in various combinations were employed to target specific nuclear types. Nuclei were isolated at d 8 of BAP differentiation of H1 human embryonic stem cells and fixed in 4 % paraformaldehyde. After permeabilization in 0.1 % triton X-100, nuclei were incubated for 3 and 1 h at 4 °C with primary and secondary antibodies respectively and nuclear samples were then subjected to FANS. By using markers identified by snRNA and immunohistochemistry, nuclei were first sorted into a Topoisomerase-1, or TOP1, bright population and then into the two STB subpopulations by using antibodies to JUNB (Jun B Proto-Oncogene) and TFCP2L1 (Transcription Factor CP2 Like 1). The protocol established here is simple, straightforward, and efficient and can be used on a relatively large scale to sort individual subtypes of nuclei from mixed populations of trophoblasts for further analysis., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this article., (Copyright © 2024 Elsevier Ltd. All rights reserved.)
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- 2024
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8. Development of properly-polarized trophoblast stem cell-derived organoids to model early human pregnancy.
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Zhou J, Sheridan MA, Tian Y, Dahlgren KJ, Messler M, Peng T, Ezashi T, Schulz LC, Ulery BD, Roberts RM, and Schust DJ
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The development of human trophoblast stem cells (hTSC) and stem cell-derived trophoblast organoids has enabled investigation of placental physiology and disease and early maternal-fetal interactions during a stage of human pregnancy that previously had been severely restricted. A key shortcoming in existing trophoblast organoid methodologies is the non-physiologic position of the syncytiotrophoblast (STB) within the inner portion of the organoid, which neither recapitulates placental villous morphology in vivo nor allows for facile modeling of STB exposure to the endometrium or the contents of the intervillous space. Here we have successfully established properly-polarized human trophoblast stem cell (hTSC)-sourced organoids with STB forming on the surface of the organoid. These organoids can also be induced to give rise to the extravillous trophoblast (EVT) lineage with HLA-G
+ migratory cells that invade into an extracellular matrix-based hydrogel. Compared to previous hTSC organoid methods, organoids created by this method more closely mimic the architecture of the developing human placenta and provide a novel platform to study normal and abnormal human placental development and to model exposures to pharmaceuticals, pathogens and environmental insults., Motivation: Human placental organoids have been generated to mimic physiological cell-cell interactions. However, those published models derived from human trophoblast stem cells (hTSCs) or placental villi display a non-physiologic "inside-out" morphology. In vivo , the placental villi have an outer layer of syncytialized cells that are in direct contact with maternal blood, acting as a conduit for gas and nutrient exchange, and an inner layer of progenitor, single cytotrophoblast cells that fuse to create the syncytiotrophoblast layer. Existing "inside-out" models put the cytotrophoblast cells in contact with culture media and substrate, making physiologic interactions between syncytiotrophoblast and other cells/tissues and normal and pathogenic exposures coming from maternal blood difficult to model. The goal of this study was to develop an hTSC-derived 3-D human trophoblast organoid model that positions the syncytiotrophoblast layer on the outside of the multicellular organoid.- Published
- 2023
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9. Lower female survival from an opportunistic infection reveals progesterone-driven sex bias in trained immunity.
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Earhart AP, Karasseva NG, Storey KM, Olthoff B, Sarker MB, Laffey KG, Lange MJ, Rector RS, Schulz LC, Gil D, Neuhauser CM, and Schrum AG
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- Female, Male, Animals, Mice, Sexism, Trained Immunity, Adaptive Immunity, Progesterone pharmacology, Opportunistic Infections
- Abstract
Immune responses differ between females and males, although such sex-based variance is incompletely understood. Observing that bacteremia of the opportunistic pathogen Burkholderia gladioli caused many more deaths of female than male mice bearing genetic deficiencies in adaptive immunity, we determined that this was associated with sex bias in the innate immune memory response called trained immunity. Female attenuation of trained immunity varies with estrous cycle stage and correlates with serum progesterone, a hormone that decreases glycolytic capacity and recall cytokine secretion induced by antigen non-specific stimuli. Progesterone receptor antagonism rescues female trained immune responses and survival from controlled B. gladioli infection to magnitudes similar to those of males. These data demonstrate progesterone-dependent sex bias in trained immunity where attenuation of female responses is associated with survival outcomes from opportunistic infection., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)
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- 2023
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10. RISING STARS: Approaches to modeling placental function in preeclampsia in vitro and in vivo.
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Milano-Foster J and Schulz LC
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- Mice, Animals, Pregnancy, Female, Humans, Coculture Techniques, Cell Culture Techniques, Ischemia, Trophoblasts, Mammals, Placenta, Pre-Eclampsia
- Abstract
Modeling preeclampsia remains difficult due to the nature of the disease and the unique characteristics of the human placenta. Members of the Hominidae superfamily have a villous hemochorial placenta that is different in structure from those of other therian mammals, including the mouse hemochorial placenta, making this common animal model less ideal for studying this disease. Human placental tissues delivered from pregnancies complicated by preeclampsia are excellent for assessing the damage the disease causes but cannot answer how or when the disease begins. Symptoms of preeclampsia manifest halfway through pregnancy or later, making it currently impossible to identify preeclampsia in human tissues obtained from an early stage of pregnancy. Many animal and cell culture models recapitulate various aspects of preeclampsia, though none can on its own completely capture the complexity of human preeclampsia. It is particularly difficult to uncover the cause of the disease using models in which the disease is induced in the lab. However, the many ways by which preeclampsia-like features can be induced in a variety of laboratory animals are consistent with the idea that preeclampsia is a two-stage disease, in which a variety of initial insults may lead to placental ischemia, and ultimately systemic symptoms. The recent development of stem cell-based models, organoids, and various coculture systems have brought in vitro systems with human cells ever closer to recapitulating in vivo events that lead to placental ischemia.
- Published
- 2023
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11. Effect of Genetically Reduced Maternal Myostatin on Late Gestation Maternal, Fetal, and Placental Metabolomes in Mice.
- Author
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Opoku R, DeCata J, Phillips CL, and Schulz LC
- Abstract
Myostatin (gene symbol: Mstn ) is an autocrine and paracrine inhibitor of muscle growth. Pregnant mice with genetically reduced levels of myostatin give birth to offspring with greater adult muscle mass and bone biomechanical strength. However, maternal myostatin is not detectable in fetal circulations. Fetal growth is dependent on the maternal environment, and the provisioning of nutrients and growth factors by the placenta. Thus, this study examined the effect of reduced maternal myostatin on maternal and fetal serum metabolomes, as well as the placental metabolome. Fetal and maternal serum metabolomes were highly distinct, which is consistent with the role of the placenta in creating a specific fetal nutrient environment. There was no effect from myostatin on maternal glucose tolerance or fasting insulin. In comparisons between pregnant control and Mstn
+/- mice, there were more significantly different metabolite concentrations in fetal serum, at 50, than in the mother's serum at 33, confirming the effect of maternal myostatin reduction on the fetal metabolic milieu. Polyamines, lysophospholipids, fatty acid oxidation, and vitamin C, in fetal serum, were all affected by maternal myostatin reduction.- Published
- 2023
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12. The immune checkpoint molecule, VTCN1/B7-H4, guides differentiation and suppresses proinflammatory responses and MHC class I expression in an embryonic stem cell-derived model of human trophoblast.
- Author
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Zhou J, Tian Y, Qu Y, Williams M, Yuan Y, Karvas RM, Sheridan MA, Schulz LC, Ezashi T, Roberts MR, and Schust DJ
- Subjects
- Infant, Newborn, Pregnancy, Humans, Female, Placenta metabolism, Immune Checkpoint Proteins metabolism, Histocompatibility Antigens Class I genetics, Histocompatibility Antigens Class I metabolism, HLA Antigens, Embryonic Stem Cells, Cell Differentiation, V-Set Domain-Containing T-Cell Activation Inhibitor 1 metabolism, Trophoblasts metabolism, Premature Birth metabolism
- Abstract
The placenta acts as a protective barrier to pathogens and other harmful substances present in the maternal circulation throughout pregnancy. Disruption of placental development can lead to complications of pregnancy such as preeclampsia, intrauterine growth retardation and preterm birth. In previous work, we have shown that expression of the immune checkpoint regulator, B7-H4/VTCN1, is increased upon differentiation of human embryonic stem cells (hESC) to an in vitro model of primitive trophoblast (TB), that VTCN1/B7-H4 is expressed in first trimester but not term human placenta and that primitive trophoblast may be uniquely susceptible to certain pathogens. Here we report on the role of VTCN1 in trophoblast lineage development and anti-viral responses and the effects of changes in these processes on major histocompatibility complex (MHC) class I expression and peripheral NK cell phenotypes., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Zhou, Tian, Qu, Williams, Yuan, Karvas, Sheridan, Schulz, Ezashi, Roberts and Schust.)
- Published
- 2023
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13. Association of serum leptin at 24-28 weeks gestation with initiation and progression of labor in women.
- Author
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Branham KKR, Sherman E, Golzy M, Drobnis EZ, and Schulz LC
- Subjects
- Body Mass Index, Female, Gestational Age, Glucose, Humans, Pregnancy, Labor, Obstetric, Leptin
- Abstract
Concentrations of the hormone leptin, which is produced by adipose tissue, increase with increasing BMI, whereas leptin sensitivity often declines with higher BMI. Thus, altered leptin signaling may play a role in reproductive health risks observed with increasing BMI, which include later onset and slow progression of labor. Conflicting evidence from clinical, animal and in vitro studies have suggested that leptin either promotes or inhibits labor. We hypothesized that serum leptin concentrations or serum leptin: body mass index (BMI) ratios in women may be associated with the initiation and progression of labor. Following informed consent, serum samples were collected from 90 women with singleton pregnancies at the time of routine glucose-challenge testing, for measurement of leptin. The potential influence of leptin on gestation length and cervical dilation timing were examined by multiple linear regression. Data were analyzed from 63 participants who met exclusion and inclusion criteria. Leptin concentrations (log-transformed) at 24-28 weeks gestation were not significantly correlated with first trimester BMI . Log serum leptin and leptin: BMI ratio each were significantly associated with shorter total gestation length in uncomplicated, term pregnancies. In contrast, the mid-pregnancy leptin concentrations were not associated with progression of labor, assessed by cervical dilation over time. The association between higher serum leptin and shorter gestation length is consistent with the hypothesis that leptin promotes, or is permissive for, the onset of labor., (© 2022. The Author(s).)
- Published
- 2022
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14. Conditional knockout of leptin receptor in the female reproductive tract reduces fertility due to parturition defects in mice.
- Author
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Pennington KA, Oestreich AK, Cataldo KH, Fogliatti CM, Lightner C, Lydon JP, and Schulz LC
- Subjects
- Animals, Embryo Implantation physiology, Female, Mice, Mice, Knockout, Parturition, Pregnancy, Uterus, Fertility genetics, Receptors, Leptin genetics
- Abstract
Leptin is required for fertility, including initiation of estrous cycles. It is therefore challenging to assess the role of leptin signaling during pregnancy. Although neuron-specific transgene approaches suggest that leptin signaling in the central nervous system is most important, experiments with pharmacologic inhibition of leptin in the uterus or global replacement of leptin during pregnancy suggest leptin signaling in the reproductive tract may be required. Here, conditional leptin receptor knockout (Lepr cKO) with a progesterone receptor-driven Cre recombinase was used to examine the importance of leptin signaling in pregnancy. Lepr cKO mice have almost no leptin receptor in uterus or cervix, and slightly reduced leptin receptor levels in corpus luteum. Estrous cycles and progesterone concentrations were not affected by Lepr cKO. Numbers of viable embryos did not differ between primiparous control and Lepr cKO dams on Days 6.5 and 17.5 of pregnancy, despite a slight reduction in the ratio of embryos to corpora lutea, showing that uterine leptin receptor signaling is not required for embryo implantation. Placentas of Lepr cKO dams had normal weight and structure. However, over four parities, Lepr cKO mice produced 22% fewer live pups than controls, and took more time from pairing to delivery by their fourth parity. Abnormal birth outcomes of either dystocia or dead pups occurred in 33% of Lepr cKO deliveries but zero control deliveries, and the average time to deliver each pup after crouching was significantly increased. Thus, leptin receptor signaling in the reproductive tract is required for normal labor and delivery., (Published by Oxford University Press on behalf of Society for the Study of Reproduction 2022.)
- Published
- 2022
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15. Placental structural abnormalities in gestational diabetes and when they develop: A scoping review.
- Author
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Ehlers E, Talton OO, Schust DJ, and Schulz LC
- Subjects
- Diabetes, Gestational diagnostic imaging, Female, Humans, Placenta diagnostic imaging, Pregnancy, Ultrasonography, Prenatal, Diabetes, Gestational pathology, Placenta pathology
- Abstract
Gestational diabetes mellitus (GDM) is defined as diabetes with onset or first recognition during gestation. It is a common complication of pregnancy that has become more prevalent over the past few decades. Abnormalities in fetal growth, including increased incidence of both large and small for gestational age babies, suggest placental dysfunction. The major goal of this scoping review is to determine what is known about abnormalities in placentas delivered from GDM pregnancies, and how early in gestation these abnormalities arise. A secondary goal is to review to what extent other selected factors, in particular obesity, have been found to influence or modify the reported effects of GDM on placental development, and whether these are considered in the study of GDM placentas. PubMed and Scopus databases were searched using the key terms: "gestational diabetes AND (woman OR human) AND placenta AND (ultrasound OR ultrastructure OR imaging OR histology OR pathology). Studies of gross morphology and histoarchitecture in placentas delivered from GDM pregnancies consistently report increased placental size, villous immaturity and a range of vascular lesions when compared to uncomplicated pregnancies. In contrast, a small number of ultrasound studies have examined placental development in GDM pregnancies in the second, and especially, the first trimester. Relatively few studies have analyzed interactions with maternal BMI, but these do suggest that it may play a role in placental abnormalities. Further examination of placental development early in pregnancy is needed to understand when it becomes disrupted in GDM, as a first step to identifying the underlying causes., (Copyright © 2021 Elsevier Ltd. All rights reserved.)
- Published
- 2021
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16. Syncytins expressed in human placental trophoblast.
- Author
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Roberts RM, Ezashi T, Schulz LC, Sugimoto J, Schust DJ, Khan T, and Zhou J
- Subjects
- Cell Fusion, Female, Gene Products, env genetics, Humans, Pregnancy, Pregnancy Proteins genetics, Biological Evolution, Endogenous Retroviruses genetics, Gene Products, env metabolism, Placentation, Pregnancy Proteins metabolism, Trophoblasts metabolism
- Abstract
Three versions of syncytiotrophoblast exist in the human placenta: an invasive type associated with the implanting conceptus, non-invasive villous type of definitive placenta, and placental bed giant cells. Syncytins are encoded by modified env genes of endogenous retroviruses (ERV), but how they contribute functionally to placental syncytial structures is unclear. A minimum of eight genes (ERVW1, ERVFRD-1, ERVV-1, ERVV-2, ERVH48-1, ERVMER34-1, ERV3-1, & ERVK13-1) encoding syncytin family members are expressed in human trophoblast, the majority from implantation to term. ERVW1 (Syncytin 1) and ERVFRD-1 (Syncytin 2) are considered the major fusogens, but, when the expression of their genes is analyzed by single cell RNAseq in first trimester placenta, their transcripts are distinctly patterned and also differ from those of their proposed binding partners, SLC1A5 and MFSD2A, respectively. ERVRH48-1 (suppressyn or SUPYN) and ERVMER34-1 are probable negative regulators of fusion and co-expressed, primarily in cytotrophoblast. The remaining genes and their products have been little studied. Syncytin expression is a feature of placental development in almost all eutherian mammals studied, in at least one marsupial, and in viviparous lizards, which lack the trophoblast lineage. Their expression has been inferred to be essential for pregnancy success in the mouse. All the main human ERV genes arose following independent retroviral insertion events, none of which trace back to the divergence of eutherians and metatherians (marsupials). While syncytins may be crucial for placental development, it seems unlikely that they helped orchestrate the divergence of eutherians and marsupials., (Copyright © 2021 Elsevier Ltd. All rights reserved.)
- Published
- 2021
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17. Modeling human peri-implantation placental development and function†.
- Author
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Zhou J, West RC, Ehlers EL, Ezashi T, Schulz LC, Roberts RM, Yuan Y, and Schust DJ
- Subjects
- Female, Humans, Pregnancy, Stem Cells metabolism, Blastocyst metabolism, Embryo Implantation, Placentation, Trophoblasts metabolism
- Abstract
It is very difficult to gain a better understanding of the events in human pregnancy that occur during and just after implantation because such pregnancies are not yet clinically detectable. Animal models of human placentation are inadequate. In vitro models that utilize immortalized cell lines and cells derived from trophoblast cancers have multiple limitations. Primary cell and tissue cultures often have limited lifespans and cannot be obtained from the peri-implantation period. We present here two contemporary models of human peri-implantation placental development: extended blastocyst culture and stem-cell derived trophoblast culture. We discuss current research efforts that employ these models and how such models might be used in the future to study the "black box" stage of human pregnancy., (Published by Oxford University Press on behalf of Society for the Study of Reproduction 2021.)
- Published
- 2021
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18. Impact of Genetic and Pharmacologic Inhibition of Myostatin in a Murine Model of Osteogenesis Imperfecta.
- Author
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Omosule CL, Gremminger VL, Aguillard AM, Jeong Y, Harrelson EN, Miloscio L, Mastaitis J, Rafique A, Kleiner S, Pfeiffer FM, Zhang A, Schulz LC, and Phillips CL
- Subjects
- Animals, Bone and Bones, Collagen Type I, Disease Models, Animal, Female, Male, Mice, Myostatin genetics, Osteogenesis, Osteogenesis Imperfecta drug therapy, Osteogenesis Imperfecta genetics
- Abstract
Osteogenesis imperfecta (OI) is a genetic connective tissue disorder characterized by compromised skeletal integrity, altered microarchitecture, and bone fragility. Current OI treatment strategies focus on bone antiresorptives and surgical intervention with limited effectiveness, and thus identifying alternative therapeutic options remains critical. Muscle is an important stimulus for bone formation. Myostatin, a TGF-β superfamily myokine, acts through ActRIIB to negatively regulate muscle growth. Recent studies demonstrated the potential benefit of myostatin inhibition with the soluble ActRIIB fusion protein on skeletal properties, although various OI mouse models exhibited variable skeletal responses. The genetic and clinical heterogeneity associated with OI, the lack of specificity of the ActRIIB decoy molecule for myostatin alone, and adverse events in human clinical trials further the need to clarify myostatin's therapeutic potential and role in skeletal integrity. In this study, we determined musculoskeletal outcomes of genetic myostatin deficiency and postnatal pharmacological myostatin inhibition by a monoclonal anti-myostatin antibody (Regn647) in the G610C mouse, a model of mild-moderate type I/IV human OI. In the postnatal study, 5-week-old wild-type and +/G610C male and female littermates were treated with Regn647 or a control antibody for 11 weeks or for 7 weeks followed by a 4-week treatment holiday. Inhibition of myostatin, whether genetically or pharmacologically, increased muscle mass regardless of OI genotype, although to varying degrees. Genetic myostatin deficiency increased hindlimb muscle weights by 6.9% to 34.4%, whereas pharmacological inhibition increased them by 13.5% to 29.6%. Female +/mstn +/G610C (Dbl.Het) mice tended to have similar trabecular and cortical bone parameters as Wt showing reversal of +/G610C characteristics but with minimal effect of +/mstn occurring in male mice. Pharmacologic myostatin inhibition failed to improve skeletal bone properties of male or female +/G610C mice, although skeletal microarchitectural and biomechanical improvements were observed in male wild-type mice. Four-week treatment holiday did not alter skeletal outcomes. © 2020 American Society for Bone and Mineral Research (ASBMR)., (© 2020 American Society for Bone and Mineral Research (ASBMR).)
- Published
- 2021
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19. Skeletal muscle specific mitochondrial dysfunction and altered energy metabolism in a murine model (oim/oim) of severe osteogenesis imperfecta.
- Author
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Gremminger VL, Harrelson EN, Crawford TK, Ohler A, Schulz LC, Rector RS, and Phillips CL
- Subjects
- Animals, Disease Models, Animal, Femur metabolism, Femur pathology, Humans, Mice, Mitochondria, Heart physiology, Muscle, Skeletal pathology, Osteogenesis Imperfecta metabolism, Osteogenesis Imperfecta pathology, Severity of Illness Index, Energy Metabolism genetics, Mitochondria, Heart genetics, Muscle, Skeletal metabolism, Osteogenesis Imperfecta genetics
- Abstract
Osteogenesis imperfecta (OI) is a heritable connective tissue disorder with patients exhibiting bone fragility and muscle weakness. The synergistic biochemical and biomechanical relationship between bone and muscle is a critical potential therapeutic target, such that muscle weakness should not be ignored. Previous studies demonstrated mitochondrial dysfunction in the skeletal muscle of oim/oim mice, which model a severe human type III OI. Here, we further characterize this mitochondrial dysfunction and evaluate several parameters of whole body and skeletal muscle metabolism. We demonstrate reduced mitochondrial respiration in female gastrocnemius muscle, but not in liver or heart mitochondria, suggesting that mitochondrial dysfunction is not global in the oim/oim mouse. Myosin heavy chain fiber type distributions were altered in the oim/oim soleus muscle with a decrease (-33 to 50%) in type I myofibers and an increase (+31%) in type IIa myofibers relative to their wildtype (WT) littermates. Additionally, altered body composition and increased energy expenditure were observed oim/oim mice relative to WT littermates. These results suggest that skeletal muscle mitochondrial dysfunction is linked to whole body metabolic alterations and to skeletal muscle weakness in the oim/oim mouse., (Copyright © 2021 Elsevier Inc. All rights reserved.)
- Published
- 2021
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20. Effects of maternal nutrient restriction during the periconceptional period on placental development in the mouse.
- Author
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Van Gronigen Case G, Storey KM, Parmeley LE, and Schulz LC
- Subjects
- Animals, DNA Methylation, Female, Fertilization, Fetal Development, Male, Mice, Nutrients deficiency, Placenta anatomy & histology, Pregnancy, Prenatal Nutritional Physiological Phenomena, Real-Time Polymerase Chain Reaction, Trophoblasts, Caloric Restriction adverse effects, Placentation physiology
- Abstract
Maternal undernutrition has detrimental effects on fetal development and adult health. Total caloric restriction during early pregnancy followed by adequate nutrition for the remainder of gestation, is particularly linked to cardiovascular and metabolic disease risks during adulthood. The placenta is responsible for transport of nutrients from the maternal to fetal circulation, and the efficiency with which it does so can be adjusted to the maternal nutrient supply. There is evidence that placental adaptations to nutrient restriction in early pregnancy may be retained even when adequate nutrition is restored later in pregnancy, leading to a potential mismatch between placental efficiency and maternal nutrient supplies. However, in the mouse, 50% caloric restriction from days 1.5-11.5 of gestation, while temporarily altering placental structure and gene expression, had no significant effect on day 18.5. The periconceptional period, during which oocyte maturation, fertilization, and preimplantation development occur may be especially critical in creating lasting impact on the placenta. Here, mice were subjected to 50% caloric restriction from 3 weeks prior to pregnancy through d11.5, and then placental structure, the expression of key nutrient transporters, and global DNA methylation levels were examined at gestation d18.5. Prior exposure to caloric restriction increased maternal blood space area, but decreased expression of the key System A amino acid transporter Slc38a4 at d18.5. Neither placental and fetal weights, nor placental DNA methylation levels were affected. Thus, total caloric restriction beginning in the periconceptional period does have a lasting impact on placental development in the mouse, but without changing placental efficiency., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2021
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21. Transcription Factor PLAGL1 Is Associated with Angiogenic Gene Expression in the Placenta.
- Author
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Starks RR, Abu Alhasan R, Kaur H, Pennington KA, Schulz LC, and Tuteja G
- Subjects
- Animals, Binding Sites, Cell Cycle Proteins metabolism, DNA-Binding Proteins metabolism, Endothelial Cells metabolism, Female, Gene Expression, Gene Regulatory Networks, Humans, Mice, Mice, Inbred C57BL, Neovascularization, Physiologic genetics, Pregnancy, Transcription Factors metabolism, Trophoblasts metabolism, Tumor Suppressor Proteins metabolism, Cell Cycle Proteins genetics, DNA-Binding Proteins genetics, Placenta blood supply, Placenta metabolism, Placentation genetics, Transcription Factors genetics, Tumor Suppressor Proteins genetics
- Abstract
During pregnancy, the placenta is important for transporting nutrients and waste between the maternal and fetal blood supply, secreting hormones, and serving as a protective barrier. To better understand placental development, we must understand how placental gene expression is regulated. We used RNA-seq data and ChIP-seq data for the enhancer associated mark, H3k27ac, to study gene regulation in the mouse placenta at embryonic day (e) 9.5, when the placenta is developing a complex network of blood vessels. We identified several upregulated transcription factors with enriched binding sites in e9.5-specific enhancers. The most enriched transcription factor, PLAGL1 had a predicted motif in 233 regions that were significantly associated with vasculature development and response to insulin stimulus genes. We then performed several experiments using mouse placenta and a human trophoblast cell line to understand the role of PLAGL1 in placental development. In the mouse placenta, Plagl1 is expressed in endothelial cells of the labyrinth layer and is differentially expressed in placentas from mice with gestational diabetes compared to placentas from control mice in a sex-specific manner. In human trophoblast cells, siRNA knockdown significantly decreased expression of genes associated with placental vasculature development terms. In a tube assay, decreased PLAGL1 expression led to reduced cord formation. These results suggest that Plagl1 regulates overlapping gene networks in placental trophoblast and endothelial cells, and may play a critical role in placental development in normal and complicated pregnancies.
- Published
- 2020
- Full Text
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22. Fecundity is impaired in a mouse model of osteogenesis imperfecta.
- Author
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Oestreich AK, DeCata JA, Akers JD, Phillips CL, and Schulz LC
- Subjects
- Animals, Collagen Type I genetics, Disease Models, Animal, Female, Fertility genetics, Fetal Viability genetics, Humans, Infertility, Female genetics, Infertility, Female pathology, Litter Size genetics, Male, Mice, Mice, Transgenic, Mutation, Osteogenesis Imperfecta genetics, Osteogenesis Imperfecta pathology, Pregnancy, Pregnancy, High-Risk genetics, Infertility, Female etiology, Osteogenesis Imperfecta complications
- Abstract
Osteogenesis imperfecta (OI), or brittle bone disease, is most often caused by mutations in genes encoding type I collagen or proteins that process it. Women with OI have a small, but significant increase in risk of serious pregnancy complications including uterine rupture. Here, the OI mouse, Col1a2
oim/oim , was used to examine the effects of collagen mutation on establishment and maintenance of pregnancy. Picrosirius birefringence was faint in Col1a2oim/oim uteri, indicating diminished collagen in the myometrium and endometrium. There was some evidence of increased uterine gland number (p = .055) and size (p = .12) in (p = .055) virgin uteri, though the they were not significantly different than controls. There were no differences in the number of corpora lutea, or the time from pairing to delivery of pups between Col1a2oim/oim and control dams, suggesting that ovulation and conception occur normally. However, when examined at Gestation Day 6.5 (postimplantation), gestation Day 10.5 (midpregnancy), and Postnatal Days 1-2, Col1a2oim/oim dams had significantly fewer viable pups than controls overall. In pairwise comparisons, the loss was only significant in the postnatal group, suggesting the gradual loss of pups over time. Overall, the Col1a2oim/oim mouse data suggest that OI impairs uterine function in pregnancy in a way that affects a small but significant number of fetuses., (© 2020 Wiley Periodicals LLC.)- Published
- 2020
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23. Leprdb/+ Dams Protect Wild-type Male Offspring Bone Strength from the Detrimental Effects of a High-Fat Diet.
- Author
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Oestreich AK, Onuzuriuke A, Yao X, Talton O, Wang Y, Pfeiffer FM, Schulz LC, and Phillips CL
- Subjects
- Animals, Bone Density drug effects, Bone Development drug effects, Bone Development genetics, Compressive Strength drug effects, Compressive Strength physiology, Dietary Fats pharmacology, Female, Male, Mice, Mice, Inbred C57BL, Mice, Transgenic, Pregnancy, Prenatal Exposure Delayed Effects metabolism, Prenatal Exposure Delayed Effects physiopathology, Sex Factors, Weight Gain drug effects, Weight Gain physiology, Bone Density genetics, Diet, High-Fat adverse effects, Prenatal Exposure Delayed Effects genetics, Receptors, Leptin genetics
- Abstract
The prevalence of maternal obesity is increasing at an alarming rate and increases the life-long risk of developing cardiometabolic disease in adult offspring. Leptin, an adipokine, is systemically elevated in the obese milieu. We recently showed that maternal hyperleptinemia without obesity improves offspring insulin sensitivity and glucose tolerance while protecting against weight gain on a high-fat, high-sugar (HFD). Here, we investigate the effect of maternal hyperleptinemia on offspring bone by using 2 independent maternal models. First, we compared wild-type (WT) offspring from severely hyperleptinemic Leprdb/+ (DB/+) dams with those from WT dams. In the second model, WT females were implanted with miniosmotic pumps that released either saline (group SAL) or leptin (group LEP; 650ng/hour) and the WT offspring were compared. At 23 weeks of age, a subset of offspring were challenged with a HFD for 8 weeks. When the offspring were 31 weeks of age, bone geometry, strength, and material properties were investigated. The HFD increased trabecular bone volume but decreased both total breaking strength and material strength of femora from the offspring of WT dams. However, male offspring of DB/+ dams were protected from the detrimental effects of a HFD, while offspring of LEP dams were not. Further material analysis revealed a modest decrease in advanced glycation end product accumulation coupled with increased collagen crosslinking in male offspring from DB/+ dams on a HFD. These data suggest that while maternal leptin may protect bone quality from the effects of a HFD, additional factors of the maternal environment controlled by leptin receptor signaling are likely also involved., (© Endocrine Society 2020. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
- Published
- 2020
- Full Text
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24. Morphology and gene expression in mouse placentas lacking leptin receptors.
- Author
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Pollock KE, Talton OO, and Schulz LC
- Subjects
- Animals, Embryo Transfer, Female, Fetal Development, Gene Expression Profiling, Male, Mice, Inbred C57BL, Pregnancy, Gene Expression Regulation, Developmental, Placenta anatomy & histology, Placenta metabolism, Receptors, Leptin deficiency
- Abstract
In the pregnant mouse, the hormone leptin is primarily produced by adipose tissue and does not significantly cross the placenta into fetal circulation. Nonetheless, leptin treatment during gestation affects offspring phenotypes. Leptin treatment also affects placental trophoblast cells in vitro, by altering proliferation, invasion and nutrient transport. The goal of the present study was to determine whether the absence of placental leptin receptors alters placental development and gene expression. Lepr
db-3j+ mice possessing only one functional copy of the leptin receptor were mated to obtain wildtype, Leprdb-3j+ and Leprdb-3j/db-3j conceptuses, which were then transferred to wildtype recipient dams. Placentas were collected at gestational d18.5 to examine placental morphology and gene expression. Placentas lacking functional leptin receptor had reduced weights, but were otherwise morphologically indistinguishable from control placentas. Relative mRNA levels, however, were altered in Leprdb-3j/db-3j placentas, particularly transcripts related to amino acid and lipid metabolism and transport. Consistent with a previous in vitro study, leptin was found to promote expression of stathmin, a positive regulator of trophoblast invasion, and of serotonin receptors, potential mediators of offspring neurological development. Overall placental leptin receptor was found not to play a significant role in morphological development of the placenta, but to regulate placental gene expression, including in metabolic pathways that affect fetal growth., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Inc. All rights reserved.)- Published
- 2020
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- View/download PDF
25. Use of a human embryonic stem cell model to discover GABRP, WFDC2, VTCN1 and ACTC1 as markers of early first trimester human trophoblast.
- Author
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Karvas RM, McInturf S, Zhou J, Ezashi T, Schust DJ, Roberts RM, and Schulz LC
- Subjects
- Actins genetics, Embryonic Stem Cells metabolism, Humans, Immunohistochemistry, Principal Component Analysis, Receptors, GABA-A genetics, Transcriptome genetics, V-Set Domain-Containing T-Cell Activation Inhibitor 1 genetics, WAP Four-Disulfide Core Domain Protein 2 genetics, Actins metabolism, Human Embryonic Stem Cells metabolism, Receptors, GABA-A metabolism, Trophoblasts metabolism, V-Set Domain-Containing T-Cell Activation Inhibitor 1 metabolism, WAP Four-Disulfide Core Domain Protein 2 metabolism
- Abstract
Human placental development during early pregnancy is poorly understood. Many conceptuses are lost at this stage. It is thought that preeclampsia, intrauterine growth restriction and other placental syndromes that manifest later in pregnancy may originate early in placentation. Thus, there is a need for models of early human placental development. Treating human embryonic stem cells (hESCs) with BMP4 (bone morphogenic protein 4) plus A83-01 (ACTIVIN/NODAL signaling inhibitor) and PD173074 (fibroblast growth factor 2 or FGF2 signaling inhibitor) (BAP conditions) induces differentiation to the trophoblast lineage (hESCBAP), but it is not clear which stage of trophoblast differentiation these cells resemble. Here, comparison of the hESCBAP transcriptome to those of trophoblasts from human blastocysts, trophoblast stem cells and placentas collected in the first-third trimester of pregnancy by principal component analysis suggests that hESC after 8 days BAP treatment most resemble first trimester syncytiotrophoblasts. To further test this hypothesis, transcripts were identified that are expressed in hESCBAP but not in cultures of trophoblasts isolated from term placentas. Proteins encoded by four genes, GABRP (gamma-aminobutyric acid type A receptor subunit Pi), WFDC2 (WAP four-disulfide core domain 2), VTCN1 (V-set domain containing T-cell activation inhibitor 1) and ACTC1 (actin alpha cardiac muscle 1), immunolocalized to placentas at 4-9 weeks gestation, and their expression declined with gestational age (R2 = 0.61-0.83). None are present at term. Expression was largely localized to syncytiotrophoblast of both hESCBAP cells and placental material from early pregnancy. WFDC2, VTCN1 and ACTC1 have not previously been described in placenta. These results support the hypothesis that hESCBAP represent human trophoblast analogous to that of early first trimester and are a tool for discovery of factors important to this stage of placentation., (© The Author(s) 2020. Published by Oxford University Press on behalf of European Society of Human Reproduction and Embryology. All rights reserved. For permissions, please email: journals.permissions@oup.com.)
- Published
- 2020
- Full Text
- View/download PDF
26. Developmental origins of ovarian disorder: impact of maternal lean gestational diabetes on the offspring ovarian proteome in mice†.
- Author
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Clark KL, Talton OO, Ganesan S, Schulz LC, and Keating AF
- Subjects
- Animals, Cell Count, Diabetes Mellitus, Experimental metabolism, Diabetes Mellitus, Experimental pathology, Diet, Female, Mice, Mice, Inbred C57BL, Ovarian Diseases metabolism, Ovarian Follicle metabolism, Ovarian Follicle pathology, Pregnancy, Prenatal Exposure Delayed Effects pathology, Proteome analysis, Thinness complications, Thinness pathology, Diabetes, Gestational metabolism, Diabetes, Gestational pathology, Ovarian Diseases etiology, Ovary metabolism, Prenatal Exposure Delayed Effects metabolism, Proteome metabolism
- Abstract
Gestational diabetes mellitus (GDM) is an obstetric disorder affecting approximately 10% of pregnancies. The four high-fat, high-sucrose (HFHS) mouse model emulates GDM in lean women. Dams are fed a HFHS diet 1 week prior to mating and throughout gestation resulting in inadequate insulin response to glucose in mid-late pregnancy. The offspring of HFHS dams have increased adiposity, thus, we hypothesized that maternal metabolic alterations during lean GDM would compromise ovarian function in offspring both basally and in response to a control or HFHS diet in adulthood. Briefly, DLPL were lean dams and control diet pups; DLPH were lean dams and HFHS pups; DHPL were HFHS dams and control diet pups; and DHPH were HFHS dams and HFHS pups. A HFHS challenge in the absence of maternal GDM (DLPL vs. DLPH) increased 3 and decreased 30 ovarian proteins. Maternal GDM in the absence of a dietary stress (DLPL vs. DHPL) increased abundance of 4 proteins and decreased abundance of 85 proteins in the offspring ovary. Finally, 87 proteins increased, and 4 proteins decreased in offspring ovaries due to dietary challenge and exposure to maternal GDM in utero (DLPL vs. DHPH). Canopy FGF signaling regulator 2, deleted in azoospermia-associated protein 1, septin 7, and serine/arginine-rich splicing factor 2 were altered across multiple offspring groups. Together, these findings suggest a possible impact on fertility and oocyte quality in relation to GDM exposure in utero as well as in response to a western diet in later life., (© The Author(s) 2018. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
- Published
- 2019
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27. Modeling the Placenta with Stem Cells.
- Author
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Ezashi T, Schust DJ, and Schulz LC
- Subjects
- Animals, Female, Humans, Pregnancy, Swine, Placenta, Stem Cells
- Published
- 2019
- Full Text
- View/download PDF
28. Changes in excitability and ion channel expression in neurons of the major pelvic ganglion in female type II diabetic mice.
- Author
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Gray M, Lett KM, Garcia VB, Kyi CW, Pennington KA, Schulz LC, and Schulz DJ
- Subjects
- Action Potentials physiology, Animals, Female, Ganglia, Sympathetic physiopathology, Ion Channels biosynthesis, Mice, Mice, Mutant Strains, Receptors, Leptin genetics, Diabetes Mellitus, Experimental metabolism, Diabetes Mellitus, Experimental physiopathology, Ganglia, Sympathetic pathology, Ion Channels metabolism, Neurons metabolism
- Abstract
Bladder cystopathy and autonomic dysfunction are common complications of diabetes, and have been associated with changes in ganglionic transmission and some measures of neuronal excitability in male mice. To determine whether type II diabetes also impacts excitability of ganglionic neurons in females, we investigated neuronal excitability and firing properties, as well as underlying ion channel expression, in major pelvic ganglion (MPG) neurons in control, 10-week, and 21-week Lepr
db/db mice. Type II diabetes in Leprdb/db animals caused a non-linear change in excitability and firing properties of MPG neurons. At 10 weeks, cells exhibited increased excitability as demonstrated by an increased likelihood of firing multiple spikes upon depolarization, decreased rebound spike latency, and overall narrower action potential half-widths as a result of increased depolarization and repolarization slopes. Conversely, at 21 weeks MPG neurons of Leprdb/db mice reversed these changes, with spiking patterns and action-potential properties largely returning to control levels. These changes are associated with numerous time-specific changes in calcium, sodium, and potassium channel subunit mRNA levels. However, Principal Components Analysis of channel expression patterns revealed that rectification of excitability is not simply a return to control levels, but rather a distinct ion channel expression profile in 21-week Leprdb/db neurons. These data indicate that type II diabetes can impact the excitability of post-ganglionic, autonomic neurons of female mice, and suggest that the non-linear progression of these properties with diabetes may be the result of compensatory changes in channel expression that act to rectify disrupted firing patterns of Leprdb/db MPG neurons., (Copyright © 2019 Elsevier B.V. All rights reserved.)- Published
- 2019
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29. Lean maternal hyperglycemia alters offspring lipid metabolism and susceptibility to diet-induced obesity in mice†.
- Author
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Talton OO, Bates K, Salazar SR, Ji T, and Schulz LC
- Subjects
- Animals, Body Composition, Diabetes, Gestational chemically induced, Dietary Carbohydrates adverse effects, Disease Susceptibility, Female, Gene Expression Regulation drug effects, Glucose Intolerance, Insulin Resistance, Male, Mice, Mice, Inbred C57BL, Pregnancy, Prenatal Exposure Delayed Effects, Diet, High-Fat adverse effects, Dietary Carbohydrates administration & dosage, Hyperglycemia, Lipid Metabolism, Obesity etiology
- Abstract
We previously developed a model of gestational diabetes mellitus (GDM) in which dams exhibit glucose intolerance, insulin resistance, and reduced insulin response to glucose challenge only during pregnancy, without accompanying obesity. Here, we aimed to determine how lean gestational glucose intolerance affects offspring risk of metabolic dysfunction. One cohort of offspring was sacrificed at 19 weeks, and one at 31 weeks, with half of the second cohort placed on a high-fat, high-sucrose diet (HFHS) at 23 weeks. Exposure to maternal glucose intolerance increased weights of HFHS-fed offspring. Chow-fed offspring of GDM dams exhibited higher body fat percentages at 4, 12, and 20 weeks of age. At 28 weeks, offspring of GDM dams fed the HFHS but not the chow diet (CD) also had higher body fat percentages than offspring of controls (CON). Exposure to GDM increased the respiratory quotient (Vol CO2/Vol O2) in offspring. Maternal GDM increased adipose mRNA levels of peroxisome proliferator-activated receptor gamma (Pparg) and adiponectin (Adipoq) in 31-week-old CD-fed male offspring, and increased mRNA levels of insulin receptor (Insr) and lipoprotein lipase (Lpl) in 31-week-old male offspring on both diets. In liver at 31 weeks, mRNA levels of peroxisome proliferator-activated receptor alpha (Ppara) were elevated in CD-fed male offspring of GDM dams, and male offspring of GDM dams exhibited higher mRNA levels of Insr on both diets. Neither fasting insulin nor glucose tolerance was affected by exposure to GDM. Our findings show that GDM comprising glucose intolerance only during pregnancy programs increased adiposity in offspring, and suggests increased insulin sensitivity of subcutaneous adipose tissue., (© The Author(s) 2019. Published by Oxford University Press on behalf of Society for the Study of Reproduction.)
- Published
- 2019
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30. Early onset preeclampsia in a model for human placental trophoblast.
- Author
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Sheridan MA, Yang Y, Jain A, Lyons AS, Yang P, Brahmasani SR, Dai A, Tian Y, Ellersieck MR, Tuteja G, Schust DJ, Schulz LC, Ezashi T, and Roberts RM
- Subjects
- Bone Morphogenetic Protein 4 metabolism, Cell Adhesion, Cell Movement, Female, Gene Expression Profiling, Gene Expression Regulation, Gene Ontology, Humans, Induced Pluripotent Stem Cells, Membrane Transport Proteins genetics, Membrane Transport Proteins metabolism, Oxidative Stress, Oxygen pharmacology, Pregnancy, Transcriptome, Placenta metabolism, Pre-Eclampsia metabolism, Trophoblasts metabolism
- Abstract
We describe a model for early onset preeclampsia (EOPE) that uses induced pluripotent stem cells (iPSCs) generated from umbilical cords of EOPE and control (CTL) pregnancies. These iPSCs were then converted to placental trophoblast (TB) representative of early pregnancy. Marker gene analysis indicated that both sets of cells differentiated at comparable rates. The cells were tested for parameters disturbed in EOPE, including invasive potential. Under 5% O
2 , CTL TB and EOPE TB lines did not differ, but, under hyperoxia (20% O2 ), invasiveness of EOPE TB was reduced. RNA sequencing analysis disclosed no consistent differences in expression of individual genes between EOPE TB and CTL TB under 20% O2 , but, a weighted correlation network analysis revealed two gene modules (CTL4 and CTL9) that, in CTL TB, were significantly linked to extent of TB invasion. CTL9, which was positively correlated with 20% O2 ( P = 0.02) and negatively correlated with invasion ( P = 0.03), was enriched for gene ontology terms relating to cell adhesion and migration, angiogenesis, preeclampsia, and stress. Two EOPE TB modules, EOPE1 and EOPE2, also correlated positively and negatively, respectively, with 20% O2 conditions, but only weakly with invasion; they largely contained the same sets of genes present in modules CTL4 and CTL9. Our experiments suggest that, in EOPE, the initial step precipitating disease is a reduced capacity of placental TB to invade caused by a dysregulation of O2 response mechanisms and that EOPE is a syndrome, in which unbalanced expression of various combinations of genes affecting TB invasion provoke disease onset., Competing Interests: The authors declare no conflict of interest., (Copyright © 2019 the Author(s). Published by PNAS.)- Published
- 2019
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31. ITGA1 is upregulated in response to oxygen over time in a BMP4 model of trophoblast.
- Author
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Karvas RM, Yang Y, Ezashi T, Schust DJ, Roberts RM, and Schulz LC
- Subjects
- Cell Differentiation drug effects, Female, Gene Expression, Human Embryonic Stem Cells metabolism, Humans, Integrin alpha1 genetics, Male, Up-Regulation, Bone Morphogenetic Protein 4 pharmacology, Integrin alpha1 metabolism, Oxygen metabolism, Trophoblasts metabolism
- Published
- 2018
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32. In Utero and Postnatal Exposure to High Fat, High Sucrose Diet Suppressed Testis Apoptosis and Reduced Sperm Count.
- Author
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Mao J, Pennington KA, Talton OO, Schulz LC, Sutovsky M, Lin Y, and Sutovsky P
- Subjects
- Animals, Animals, Newborn, Dietary Sucrose administration & dosage, Female, Male, Mice, Mice, Inbred C57BL, Pregnancy, Prenatal Exposure Delayed Effects etiology, Sperm Count, Testis drug effects, Apoptosis drug effects, Diet, High-Fat adverse effects, Dietary Sucrose adverse effects, Prenatal Exposure Delayed Effects pathology, Spermatogenesis drug effects, Testis pathology
- Abstract
Obesity affects male fertility and maternal diabetes affects the offspring sperm epigenome. However, the effects of in utero exposure to maternal glucose intolerance in combination with postnatal high fat, high sucrose (HFHS) diet consumption on offspring spermatogenesis is not clear. The present study was designed to test these effects. One week before and during pregnancy, dams were fed either control or HFHS diet to induce gestational glucose intolerance, and returned to standard diet during lactation. Male offspring from each maternal group were split into control and HFHS-fed groups for eight weeks prior to sacrifice at 11, 19 or 31 weeks of age, and reproductive tissues were harvested for analysis of testicular germ cell apoptosis and sperm output. Postnatal HFHS diet suppressed spermatogonia apoptosis in all age groups and maternal HFHS diet reduced testosterone levels at 11 weeks. At 31 weeks of age, the postnatal HFHS diet increased body weight, and reduced epididymis weight and sperm count. The combination of in utero and postnatal exposure impacted sperm counts most significantly. In summary, HFHS diet during pregnancy puts male offspring at greater risk of infertility, particularly when combined with postnatal high fat diet feeding.
- Published
- 2018
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33. Effects of acute exposure to a high-fat, high-sucrose diet on gestational glucose tolerance and subsequent maternal health in mice.
- Author
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Pennington KA, van der Walt N, Pollock KE, Talton OO, and Schulz LC
- Subjects
- Animal Feed analysis, Animal Nutritional Physiological Phenomena, Animals, Female, Glucose Intolerance, Maternal Nutritional Physiological Phenomena, Mice, Mice, Inbred C57BL, Pregnancy, Diabetes, Gestational, Diet, High-Fat adverse effects, Dietary Sucrose administration & dosage, Dietary Sucrose adverse effects, Disease Models, Animal
- Abstract
Gestational diabetes mellitus (GDM) is a common obstetric complication. Half of women who have GDM will go on to develop type 2 diabetes. Understanding the mechanisms by which this occurs requires an animal model of GDM without ongoing diabetes at conception. C57Bl/6J mice react acutely to a high-fat, high-sucrose (HFHS) challenge. Here, we hypothesized that a periconceptional HFHS challenge will induce glucose intolerance during gestation. C57Bl/6J female mice were placed on an HFHS either 1 or 3 weeks prior to mating and throughout pregnancy. Intraperitoneal glucose tolerance tests, insulin measurements, and histological analysis of pancreatic islets were used to assess the impact of acute HFHS. C57Bl/6J females fed HFHS beginning 1 week prior to pregnancy became severely glucose intolerant, with reduced insulin response to glucose, and decreased pancreatic islet expansion during pregnancy compared to control mice. These GDM characteristics did not occur when the HFHS diet was started 3 weeks prior to mating, suggesting the importance of acute metabolic stress. Additionally, HFHS feeding resulted in only mild insulin resistance in nonpregnant females. When the diet was discontinued at parturition, symptoms resolved within 3 weeks. However, mice that experienced glucose intolerance in pregnancy became glucose intolerant more readily in response to a HFHS challenge later in life than congenic females that experienced a normal pregnancy, or that were fed the same diet outside of pregnancy. Thus, acute HFHS challenge in C57Bl/6 mice results in a novel, nonobese, animal model that recapitulates the long-term risk of developing type 2 diabetes following GDM.
- Published
- 2017
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34. Decreasing maternal myostatin programs adult offspring bone strength in a mouse model of osteogenesis imperfecta.
- Author
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Oestreich AK, Kamp WM, McCray MG, Carleton SM, Karasseva N, Lenz KL, Jeong Y, Daghlas SA, Yao X, Wang Y, Pfeiffer FM, Ellersieck MR, Schulz LC, and Phillips CL
- Subjects
- Animals, Biomarkers blood, Biomechanical Phenomena, Body Weight, Collagen metabolism, Disease Models, Animal, Embryo Implantation, Female, Femur pathology, Male, Mice, Inbred C57BL, Muscle Contraction, Myostatin deficiency, Osteoblasts metabolism, Osteogenesis Imperfecta blood, Osteogenesis Imperfecta embryology, Tibia pathology, Tibia physiopathology, Femur physiopathology, Myostatin metabolism, Osteogenesis Imperfecta physiopathology
- Abstract
During fetal development, the uterine environment can have effects on offspring bone architecture and integrity that persist into adulthood; however, the biochemical and molecular mechanisms remain unknown. Myostatin is a negative regulator of muscle mass. Parental myostatin deficiency (Mstn
tm1Sjl/+ ) increases muscle mass in wild-type offspring, suggesting an intrauterine programming effect. Here, we hypothesized that Mstntm1Sjl/+ dams would also confer increased bone strength. In wild-type offspring, maternal myostatin deficiency altered fetal growth and calvarial collagen content of newborn mice and conferred a lasting impact on bone geometry and biomechanical integrity of offspring at 4 mo of age, the age of peak bone mass. Second, we sought to apply maternal myostatin deficiency to a mouse model with osteogenesis imperfecta (Col1a2oim ), a heritable connective tissue disorder caused by abnormalities in the structure and/or synthesis of type I collagen. Femora of male Col1a2oim/+ offspring from natural mating of Mstntm1Sjl/+ dams to Col1a2oim/+ sires had a 15% increase in torsional ultimate strength, a 29% increase in tensile strength, and a 24% increase in energy to failure compared with age, sex, and genotype-matched offspring from natural mating of Col1a2oim/+ dams to Col1a2oim/+ sires. Finally, increased bone biomechanical strength of Col1a2oim/+ offspring that had been transferred into Mstntm1Sjl/+ dams as blastocysts demonstrated that the effects of maternal myostatin deficiency were conferred by the postimplantation environment. Thus, targeting the gestational environment, and specifically prenatal myostatin pathways, provides a potential therapeutic window and an approach for treating osteogenesis imperfecta., Competing Interests: The authors declare no conflict of interest.- Published
- 2016
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35. The evolution of the placenta.
- Author
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Roberts RM, Green JA, and Schulz LC
- Subjects
- Animals, Female, Humans, Pregnancy, Biological Evolution, Placenta cytology, Placentation
- Abstract
The very apt definition of a placenta is coined by Mossman, namely apposition or fusion of the fetal membranes to the uterine mucosa for physiological exchange. As such, it is a specialized organ whose purpose is to provide continuing support to the developing young. By this definition, placentas have evolved within every vertebrate class other than birds. They have evolved on multiple occasions, often within quite narrow taxonomic groups. As the placenta and the maternal system associate more intimately, such that the conceptus relies extensively on maternal support, the relationship leads to increased conflict that drives adaptive changes on both sides. The story of vertebrate placentation, therefore, is one of convergent evolution at both the macromolecular and molecular levels. In this short review, we first describe the emergence of placental-like structures in nonmammalian vertebrates and then transition to mammals themselves. We close the review by discussing the mechanisms that might have favored diversity and hence evolution of the morphology and physiology of the placentas of eutherian mammals., Competing Interests: Declaration of interest The authors declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported., (© 2016 Society for Reproduction and Fertility.)
- Published
- 2016
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36. Maternal Hyperleptinemia Improves Offspring Insulin Sensitivity in Mice.
- Author
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Talton OO, Pennington KA, Pollock KE, Bates K, Ma L, Ellersieck MR, and Schulz LC
- Subjects
- Animals, Female, Insulin metabolism, Leptin pharmacology, Liver metabolism, Mice, Mutation, Pancreas metabolism, Pregnancy, Receptors, Leptin genetics, Triglycerides metabolism, Insulin Resistance physiology, Leptin blood, Non-alcoholic Fatty Liver Disease metabolism, Prenatal Exposure Delayed Effects metabolism, Receptors, Leptin metabolism
- Abstract
Maternal obesity and gestational diabetes are prevalent worldwide. Offspring of mothers with these conditions weigh more and are predisposed to metabolic syndrome. A hallmark of both conditions is maternal hyperleptinemia, but the role of elevated leptin levels during pregnancy on developmental programming is largely unknown. We previously found that offspring of hyperleptinemic mothers weighed less and had increased activity. The goal of this study was to determine whether maternal leptin affects offspring insulin sensitivity by investigating offspring glucose metabolism and lipid accumulation. Offspring from two maternal hyperleptinemic models were compared. The first model of hyperleptinemia is the Lepr(db/+) mouse, which has a mutation in one copy of the gene that encodes the leptin receptor, resulting in a truncated long form of the receptor, and hyperleptinemia. Wild-type females served as the control for the Lepr(db/+) females. For the second hyperleptinemic model, wild-type females were implanted with miniosmotic pumps, which released leptin (350 ng/h) or saline (as the control) just prior to mating and throughout gestation. In the offspring of these dams, we measured glucose tolerance; serum leptin, insulin, and triglyceride levels; liver triglycerides; pancreatic α- and β-cell numbers; body composition; incidence of nonalcoholic fatty liver disease; and the expression of key metabolic genes in the liver and adipose tissue. We found that the offspring of hyperleptinemic dams exhibited improved glucose tolerance, reduced insulin and leptin concentrations, reduced liver triglycerides, and a lower incidence of nonalcoholic fatty liver disease. Overall, maternal hyperleptinemia was beneficial for offspring glucose and lipid metabolism.
- Published
- 2016
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37. Maternal Hyperleptinemia Is Associated with Male Offspring's Altered Vascular Function and Structure in Mice.
- Author
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Pennington KA, Ramirez-Perez FI, Pollock KE, Talton OO, Foote CA, Reyes-Aldasoro CC, Wu HH, Ji T, Martinez-Lemus LA, and Schulz LC
- Subjects
- Acetylcholine, Animals, Blood Pressure, Disease Models, Animal, Female, Fibrosis, Insulin, Leptin genetics, Lipid Metabolism, Male, Mice, Pregnancy, Sex Factors, Vascular Resistance, Leptin blood, Maternal Exposure adverse effects, Prenatal Exposure Delayed Effects, Vascular Diseases etiology, Vascular Diseases pathology, Vascular Diseases physiopathology
- Abstract
Children of mothers with gestational diabetes have greater risk of developing hypertension but little is known about the mechanisms by which this occurs. The objective of this study was to test the hypothesis that high maternal concentrations of leptin during pregnancy, which are present in mothers with gestational diabetes and/or obesity, alter blood pressure, vascular structure and vascular function in offspring. Wildtype (WT) offspring of hyperleptinemic, normoglycemic, Leprdb/+ dams were compared to genotype matched offspring of WT-control dams. Vascular function was assessed in male offspring at 6, and at 31 weeks of age after half the offspring had been fed a high fat, high sucrose diet (HFD) for 6 weeks. Blood pressure was increased by HFD but not affected by maternal hyperleptinemia. On a standard diet, offspring of hyperleptinemic dams had outwardly remodeled mesenteric arteries and an enhanced vasodilatory response to insulin. In offspring of WT but not Leprdb/+ dams, HFD induced vessel hypertrophy and enhanced vasodilatory responses to acetylcholine, while HFD reduced insulin responsiveness in offspring of hyperleptinemic dams. Offspring of hyperleptinemic dams had stiffer arteries regardless of diet. Therefore, while maternal hyperleptinemia was largely beneficial to offspring vascular health under a standard diet, it had detrimental effects in offspring fed HFD. These results suggest that circulating maternal leptin concentrations may interact with other factors in the pre- and post -natal environments to contribute to altered vascular function in offspring of diabetic pregnancies.
- Published
- 2016
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38. Placental development in a mouse model of spinal muscular atrophy.
- Author
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Van Gronigen Caesar G, Dale JM, Osman EY, Garcia ML, Lorson CL, and Schulz LC
- Subjects
- Animals, Female, Male, Mice, Pregnancy, Muscular Atrophy, Spinal pathology, Muscular Atrophy, Spinal physiopathology, Placentation, SMN Complex Proteins metabolism, Trophoblasts metabolism, Trophoblasts pathology
- Abstract
Spinal Muscular Atrophy (SMA) is an autosomal recessive disorder, leading to fatal loss of motor neurons. It is caused by loss of function of the SMN gene, which is expressed throughout the body, and there is increasing evidence of dysfunction in non-neuronal tissues. Birthweight is one of most powerful prognostic factors for infants born with SMA, and intrauterine growth restriction is common. In the SMNΔ7 mouse model of SMA, pups with the disease lived 25% longer when their mothers were fed a higher fat, "breeder" diet. The placenta is responsible for transport of nutrients from mother to fetus, and is a major determinant of fetal growth. Thus, the present study tested the hypothesis that placental development is impaired in SMNΔ7 conceptuses. Detailed morphological characterization revealed no defects in SMNΔ7 placental development, and expression of key transcription factors regulating mouse placental development was unaffected. The intrauterine growth restriction observed in SMA infants likely does not result from impaired placental development., (Copyright © 2016 Elsevier Inc. All rights reserved.)
- Published
- 2016
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39. Hyperleptinemia During Pregnancy Decreases Adult Weight of Offspring and Is Associated With Increased Offspring Locomotor Activity in Mice.
- Author
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Pollock KE, Stevens D, Pennington KA, Thaisrivongs R, Kaiser J, Ellersieck MR, Miller DK, and Schulz LC
- Subjects
- Animals, Body Weight drug effects, Body Weight genetics, Diet, High-Fat, Eating drug effects, Eating genetics, Exploratory Behavior drug effects, Female, Gene Expression, Leptin pharmacology, Male, Metabolic Diseases blood, Metabolic Diseases physiopathology, Mice, Inbred C57BL, Mice, Knockout, Motor Activity drug effects, Motor Activity genetics, Mutation, Obesity etiology, Obesity genetics, Obesity physiopathology, Pregnancy, Pregnancy Complications blood, Receptors, Leptin genetics, Receptors, Leptin metabolism, Reverse Transcriptase Polymerase Chain Reaction, Time Factors, Body Weight physiology, Leptin blood, Motor Activity physiology, Pregnancy Complications physiopathology
- Abstract
Pregnant women who are obese or have gestational diabetes mellitus have elevated leptin levels and their children have an increased risk for child and adult obesity. The goals of this study were to determine whether offspring weights are altered by maternal hyperleptinemia, and whether this occurs via behavioral changes that influence energy balance. We used 2 hyperleptinemic mouse models. The first was females heterozygous for a leptin receptor mutation (DB/+), which were severely hyperleptinemic, and that were compared with wild-type females. The second model was wild-type females infused with leptin (LEP), which were moderately hyperleptinemic, and were compared with wild-type females infused with saline (SAL). Total food consumption, food preference, locomotor activity, coordinated motor skills, and anxiety-like behaviors were assessed in wild-type offspring from each maternal group at 3 postnatal ages: 4-6, 11-13, and 19-21 weeks. Half the offspring from each group were then placed on a high-fat diet, and behaviors were reassessed. Adult offspring from both groups of hyperleptinemic dams weighed less than their respective controls beginning at 23 weeks of age, independent of diet or sex. Weight differences were not explained by food consumption or preference, because female offspring from hyperleptinemic dams tended to consume more food and had reduced preference for palatable, high-fat and sugar, food compared with controls. Offspring from DB/+ dams were more active than offspring of controls, as were female offspring of LEP dams. Maternal hyperleptinemia during pregnancy did not predispose offspring to obesity, and in fact, reduced weight gain.
- Published
- 2015
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40. Placental changes caused by food restriction during early pregnancy in mice are reversible.
- Author
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Harper JL, Caesar GA, Pennington KA, Davis JW, and Schulz LC
- Subjects
- Animals, Female, Gene Expression Regulation, Germinal Center Kinases, Gestational Age, Male, Mice, PPAR alpha metabolism, Pregnancy, Protein Serine-Threonine Kinases metabolism, Caloric Restriction adverse effects, Liver metabolism, Placenta metabolism, Placenta pathology, Transcriptome
- Abstract
In a previous study, 50% calorie restriction in mice from d1.5 to 11.5 of pregnancy resulted in reduced placental weights and areas,relative sparing of labyrinth zone area compared to junctional zone area, and dramatic changes in global gene expression profiles.However, little lasting effect was seen on adult offspring of these pregnancies, with a slight reduction in adiposity in males and some changes in liver gene expression in both sexes. The goals of the present study were to determine whether the placental changes induced by caloric restriction in early pregnancy had permanent, irreversible effects on the placenta, and whether the changes in liver gene expression in adult offspring were present before birth. There were no differences in placental weights or areas, or the areas of individual placental zones near term in mice that had previously been food restricted. Global gene expression profiles at d18.5 were indistinguishable in placentas from control and previously food-restricted mothers. In fetuses from restricted dams at d18.5, liver expression of Gck, a key regulator of glycogen synthesis, was reduced, whereas its expression was increased in livers from adult offspring of restricted dams. Ppara expression was also reduced in fetal livers from restricted dams at d18.5, but not in adult offspring livers. We conclude that alterations in the placenta caused by nutrient restriction in early pregnancy are reversible, and that alterations in gene expression in livers of adult offspring are not a result of changes initiated during pregnancy and maintained through adulthood.
- Published
- 2015
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41. Heightened potency of human pluripotent stem cell lines created by transient BMP4 exposure.
- Author
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Yang Y, Adachi K, Sheridan MA, Alexenko AP, Schust DJ, Schulz LC, Ezashi T, and Roberts RM
- Subjects
- Animals, Cell Differentiation, Cell Line, Cells, Cultured, Collagen chemistry, Culture Media chemistry, Culture Media, Conditioned, Drug Combinations, Embryonic Stem Cells metabolism, Female, Humans, Induced Pluripotent Stem Cells metabolism, Laminin chemistry, Mice, Oligonucleotide Array Sequence Analysis, Phenotype, Placenta metabolism, Pregnancy, Proteoglycans chemistry, Signal Transduction, Teratoma, Transcriptome, Trophoblasts metabolism, Bone Morphogenetic Protein 4 pharmacology, Gene Expression Regulation, Developmental, Pluripotent Stem Cells cytology
- Abstract
Human pluripotent stem cells (PSCs) show epiblast-type pluripotency that is maintained with ACTIVIN/FGF2 signaling. Here, we report the acquisition of a unique stem cell phenotype by both human ES cells (hESCs) and induced pluripotent stem cells (iPSCs) in response to transient (24-36 h) exposure to bone morphogenetic protein 4 (BMP4) plus inhibitors of ACTIVIN signaling (A83-01) and FGF2 (PD173074), followed by trypsin dissociation and recovery of colonies capable of growing on a gelatin substratum in standard medium for human PSCs at low but not high FGF2 concentrations. The self-renewing cell lines stain weakly for CDX2 and strongly for NANOG, can be propagated clonally on either Matrigel or gelatin, and are morphologically distinct from human PSC progenitors on either substratum but still meet standard in vitro criteria for pluripotency. They form well-differentiated teratomas in immune-compromised mice that secrete human chorionic gonadotropin (hCG) into the host mouse and include small areas of trophoblast-like cells. The cells have a distinct transcriptome profile from the human PSCs from which they were derived (including higher expression of NANOG, LEFTY1, and LEFTY2). In nonconditioned medium lacking FGF2, the colonies spontaneously differentiated along multiple lineages, including trophoblast. They responded to PD173074 in the absence of both FGF2 and BMP4 by conversion to trophoblast, and especially syncytiotrophoblast, whereas an A83-01/PD173074 combination favored increased expression of HLA-G, a marker of extravillous trophoblast. Together, these data suggest that the cell lines exhibit totipotent potential and that BMP4 can prime human PSCs to a self-renewing alternative state permissive for trophoblast development. The results may have implications for regulation of lineage decisions in the early embryo.
- Published
- 2015
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42. Abnormal oxidative stress responses in fibroblasts from preeclampsia infants.
- Author
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Yang P, Dai A, Alexenko AP, Liu Y, Stephens AJ, Schulz LC, Schust DJ, Roberts RM, and Ezashi T
- Subjects
- Case-Control Studies, Cells, Cultured, Female, Fibroblasts drug effects, Gene Expression Profiling, Glucose pharmacology, Humans, Infant, Newborn, Male, Oxygen pharmacology, Pre-Eclampsia genetics, Pre-Eclampsia metabolism, Pregnancy, Stress, Physiological drug effects, Stress, Physiological genetics, Transcriptome drug effects, Umbilical Cord metabolism, Fibroblasts metabolism, Fibroblasts pathology, Oxidative Stress drug effects, Oxidative Stress genetics, Pre-Eclampsia pathology, Umbilical Cord pathology
- Abstract
Background: Signs of severe oxidative stress are evident in term placentae of infants born to mothers with preeclampsia (PE), but it is unclear whether this is a cause or consequence of the disease. Here fibroblast lines were established from umbilical cords (UC) delivered by mothers who had experienced early onset PE and from controls with the goal of converting these primary cells to induced pluripotent stem cells and ultimately trophoblast. Contrary to expectations, the oxidative stress responses of these non-placental cells from PE infants were more severe than those from controls., Methods and Findings: Three features suggested that UC-derived fibroblasts from PE infants responded less well to oxidative stressors than controls: 1) While all UC provided outgrowths in 4% O2, success was significantly lower for PE cords in 20% O2; 2) PE lines established in 4% O2 proliferated more slowly than controls when switched to 20% O2; 3) PE lines were more susceptible to the pro-oxidants diethylmaleate and tert-butylhydroquinone than control lines, but, unlike controls, were not protected by glutathione. Transcriptome profiling revealed only a few genes differentially regulated between PE lines and controls in 4% O2 conditions. However, a more severely stressed phenotype than controls, particularly in the unfolded protein response, was evident when PE lines were switched suddenly to 20% O2, thus confirming the greater sensitivity of the PE fibroblasts to acute changes in oxidative stress., Conclusions: UC fibroblasts derived from PE infants are intrinsically less able to respond to acute oxidative stress than controls, and this phenotype is retained over many cell doublings. Whether the basis of this vulnerability is genetic or epigenetic and how it pertains to trophoblast development remains unclear, but this finding may provide a clue to the basis of the early onset, usually severe, form of PE.
- Published
- 2014
- Full Text
- View/download PDF
43. Differentiation of trophoblast cells from human embryonic stem cells: to be or not to be?
- Author
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Roberts RM, Loh KM, Amita M, Bernardo AS, Adachi K, Alexenko AP, Schust DJ, Schulz LC, Telugu BP, Ezashi T, and Pedersen RA
- Subjects
- Cell Lineage, Cells, Cultured, Embryonic Stem Cells physiology, Female, Humans, In Vitro Techniques, Morphogenesis physiology, Placenta cytology, Placenta physiology, Pregnancy, Trophoblasts physiology, Cell Differentiation physiology, Embryonic Stem Cells cytology, Trophoblasts cytology
- Abstract
It is imperative to unveil the full range of differentiated cell types into which human pluripotent stem cells (hPSCs) can develop. The need is twofold: it will delimit the therapeutic utility of these stem cells and is necessary to place their position accurately in the developmental hierarchy of lineage potential. Accumulated evidence suggested that hPSC could develop in vitro into an extraembryonic lineage (trophoblast (TB)) that is typically inaccessible to pluripotent embryonic cells during embryogenesis. However, whether these differentiated cells are truly authentic TB has been challenged. In this debate, we present a case for and a case against TB differentiation from hPSCs. By analogy to other differentiation systems, our debate is broadly applicable, as it articulates higher and more challenging standards for judging whether a given cell type has been genuinely produced from hPSC differentiation.
- Published
- 2014
- Full Text
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44. Comparison of extravillous trophoblast cells derived from human embryonic stem cells and from first trimester human placentas.
- Author
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Telugu BP, Adachi K, Schlitt JM, Ezashi T, Schust DJ, Roberts RM, and Schulz LC
- Subjects
- Bone Morphogenetic Protein 4 pharmacology, Cell Differentiation, Embryonic Stem Cells drug effects, Female, HLA-G Antigens biosynthesis, Humans, Keratin-7 biosynthesis, Oxygen pharmacology, Pregnancy, Pregnancy Trimester, First, Cell Movement physiology, Embryonic Stem Cells cytology, Placenta cytology, Trophoblasts cytology
- Abstract
Introduction: Preeclampsia and other placental pathologies are characterized by a lack of spiral artery remodeling associated with insufficient invasion by extravillous trophoblast cells (EVT). Because trophoblast invasion occurs in early pregnancy when access to human placental tissue is limited, there is a need for model systems for the study of trophoblast differentiation and invasion. Human embryonic stem cells (hESC) treated with BMP4- differentiate to trophoblast, and express HLA-G, a marker of EVT. The goals of the present study were to further characterize the HLA-G(+) cells derived from BMP4-treated hESC, and determine their suitability as a model., Methods: HESC were treated with BMP4 under 4% or 20% oxygen and tested in Matrigel invasion chambers. Both BMP4-treated hESC and primary human placental cells were separated into HLA-G(+) and HLA-G(-)/TACSTD2(+) populations with immunomagnetic beads and expression profiles analyzed by microarray., Results: There was a 10-fold increase in invasion when hESC were BMP4-treated. There was also an independent, stimulatory effect of oxygen on this process. Invasive cells expressed trophoblast marker KRT7, and the majority were also HLA-G(+). Gene expression profiles revealed that HLA-G(+), BMP4-treated hESC were similar to, but distinct from, HLA-G(+) cells isolated from first trimester placentas. Whereas HLA-G(+) and HLA-G(-) cells from first trimester placentas had highly divergent gene expression profiles, HLA-G(+) and HLA-G(-) cells from BMP4-treated hESC had somewhat similar profiles, and both expressed genes characteristic of early trophoblast development., Conclusions: We conclude that hESC treated with BMP4 provide a model for studying transition to the EVT lineage., (Copyright © 2013 Elsevier Ltd. All rights reserved.)
- Published
- 2013
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45. Complete and unidirectional conversion of human embryonic stem cells to trophoblast by BMP4.
- Author
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Amita M, Adachi K, Alexenko AP, Sinha S, Schust DJ, Schulz LC, Roberts RM, and Ezashi T
- Subjects
- Activins metabolism, Animals, Antigens, Differentiation biosynthesis, Bone Morphogenetic Protein 4 metabolism, Cell Differentiation physiology, Cell Line, Culture Media, Conditioned, Embryonic Stem Cells cytology, Fibroblast Growth Factor 2 metabolism, Humans, Keratin-7 biosynthesis, Mice, Signal Transduction physiology, Trophoblasts cytology, Bone Morphogenetic Protein 4 pharmacology, Cell Differentiation drug effects, Embryonic Stem Cells metabolism, Trophoblasts metabolism
- Abstract
Human ES cells (hESC) exposed to bone morphogenic protein 4 (BMP4) in the absence of FGF2 have become widely used for studying trophoblast development, but the soundness of this model has been challenged by others, who concluded that differentiation was primarily toward mesoderm rather than trophoblast. Here we confirm that hESC grown under the standard conditions on a medium conditioned by mouse embryonic fibroblasts in the presence of BMP4 and absence of FGF2 on a Matrigel substratum rapidly convert to an epithelium that is largely KRT7(+) within 48 h, with minimal expression of mesoderm markers, including T (Brachyury). Instead, they begin to express a series of trophoblast markers, including HLA-G, demonstrate invasive properties that are independent of the continued presence of BMP4 in the medium, and, over time, produce extensive amounts of human chorionic gonadotropin, progesterone, placental growth factor, and placental lactogen. This process of differentiation is not dependent on conditioning of the medium by mouse embryonic fibroblasts and is accelerated in the presence of inhibitors of Activin and FGF2 signaling, which at day 2 provide colonies that are entirely KRT7(+) and in which the majority of cells are transiently CDX2(+). Colonies grown on two chemically defined media, including the one in which BMP4 was reported to drive mesoderm formation, also differentiate at least partially to trophoblast in response to BMP4. The experiments demonstrate that the in vitro BMP4/hESC model is valid for studying the emergence and differentiation of trophoblasts.
- Published
- 2013
- Full Text
- View/download PDF
46. Leptin and the placental response to maternal food restriction during early pregnancy in mice.
- Author
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Schulz LC, Schlitt JM, Caesar G, and Pennington KA
- Subjects
- Animal Nutritional Physiological Phenomena, Animals, Female, Fetal Weight, Gestational Age, Leptin administration & dosage, Leptin blood, Malnutrition complications, Malnutrition pathology, Malnutrition physiopathology, Maternal Nutritional Physiological Phenomena, Mice, Microarray Analysis, Organ Size, Placenta pathology, Pregnancy, Pregnancy Complications pathology, Pregnancy Complications physiopathology, Transcriptome, Food Deprivation physiology, Leptin physiology, Placenta physiology
- Abstract
Several studies have demonstrated that maternal undernutrition or overnutrition during pregnancy can have negative consequences for the health of children born to these pregnancies, but the physiological mechanisms by which this occurs are not completely understood. During periods of food restriction, concentrations of leptin decline, whereas leptin is elevated in obesity, suggesting that it may play a role in the response to altered nutrition during pregnancy. This study compares placental development and global placental gene expression profiles at Day 11.5 in pregnant control mice, mice that were undernourished, and mice that were undernourished but given leptin. Placentas from mothers exposed to food restriction preserved the placental labyrinth zone at the expense of the junctional zone, an effect abrogated in the restricted plus leptin group, which had a significant decrease in the labyrinth zone area compared with controls. Similarly, there were more significant differences in gene expression between placentas from control and restricted plus leptin mothers (1128 differentially expressed genes) than between placentas of control and restricted mothers (281 differentially expressed genes). We conclude that the presence of high concentrations of circulating leptin during food restriction disrupts the normal adaptive response of the placenta to reduced energy availability.
- Published
- 2012
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47. Effect of food restriction and leptin supplementation on fetal programming in mice.
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Pennington KA, Harper JL, Sigafoos AN, Beffa LM, Carleton SM, Phillips CL, and Schulz LC
- Subjects
- Animals, Female, Mice, Pregnancy, Fetal Development drug effects, Food Deprivation physiology, Leptin pharmacology
- Abstract
Metabolic disease is a significant global health and economic problem. In a phenomenon referred to as fetal programming, offspring of underweight or overweight mothers have an increased incidence of adulthood obesity and metabolic disease. Undernourished individuals have decreased levels of leptin, a regulator of energy balance, whereas obese people develop hyperleptinemia and leptin resistance. We hypothesize that alterations in circulating leptin during pregnancy contribute to programming events caused by maternal nutritional status. To test this hypothesis, pregnant mice were randomly placed in one of three treatment groups: ad libitum feed plus saline injection (control, n = 5), 50% food restriction plus saline injection (restricted, n = 4), or 50% food restriction plus 1 mg/kg · d leptin injection (restricted, leptin treated, n = 4). Mice were treated from 1.5 to 11.5 d after conception and then returned to ad libitum feeding until weaning. At 19 wk after weaning, offspring were placed on a 45% fat diet and then followed up until 26 wk after weaning, at which time they were killed, and samples were collected for further analysis. Our results demonstrate that males are more negatively impacted by high-fat diet than females, regardless of maternal treatment. We provide evidence that differential response to leptin may mediate the sexual dimorphism observed in fetal programming in which male offspring are more affected by maternal undernutrition and female offspring by maternal overnutrition. We show that female offspring born to food-restricted, leptin-supplemented mothers are obese and insulin resistant. This may mimic fetal programming events seen in offspring of overweight women.
- Published
- 2012
- Full Text
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48. The source of leptin, but not leptin depletion in response to food restriction, changes during early pregnancy in mice.
- Author
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Schlitt JM and Schulz LC
- Subjects
- Animals, Blood Glucose analysis, Circadian Rhythm, Female, Gene Expression Regulation, Insulin blood, Leptin blood, Leptin genetics, Mice, Mice, Inbred Strains, Organ Specificity, Pregnancy, RNA, Messenger metabolism, Random Allocation, Real-Time Polymerase Chain Reaction, Reverse Transcriptase Polymerase Chain Reaction, Subcutaneous Fat metabolism, Caloric Restriction adverse effects, Intra-Abdominal Fat metabolism, Leptin biosynthesis, Prenatal Nutritional Physiological Phenomena
- Abstract
Maternal food restriction during pregnancy results in adverse consequences for offspring, including obesity and cardiovascular disease. Early pregnancy is a critical period for this programming effect. Leptin is a regulator of energy homeostasis that also affects placental and fetal development. As food restriction results in decreased serum leptin levels, at least in non-pregnant animals, leptin depletion may be one mechanism by which food restriction affects development. The objective of this study was to test whether moderate food restriction affects serum leptin concentrations during the first half of pregnancy. We found that restriction to 50% of ad libitum consumption levels resulted in a significant decrease in serum leptin concentrations in both pregnant and non-pregnant female mice. There was no significant difference in serum leptin concentrations between non-pregnant females and at pregnancy day 11.5 when fed ad libitum. However, there was a difference in the source of leptin during pregnancy, with greater production in visceral fat in pregnant mice, and greater production in subcutaneous fat in non-pregnant mice. Leptin concentrations were dependent on time of day and time of sampling relative to feeding, particularly in restricted mice. There was a significant difference in serum leptin concentrations between fed and restricted mice when they were fed and sampled in afternoon, but not when they were fed and sampled in morning. We conclude that food restriction results in a significant decrease in leptin concentration during the first half of pregnancy in mice, but that detection of this relationship is subject to experimental design considerations.
- Published
- 2012
- Full Text
- View/download PDF
49. Isolation of primary mouse trophoblast cells and trophoblast invasion assay.
- Author
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Pennington KA, Schlitt JM, and Schulz LC
- Subjects
- Animals, Dissection, Female, Mice, Placenta cytology, Placenta surgery, Pregnancy, Cytological Techniques methods, Trophoblasts cytology
- Abstract
The placenta is responsible for the transport of nutrients, gasses and growth factors to the fetus, as well as the elimination of wastes. Thus, defects in placental development have important consequences for the fetus and mother, and are a major cause of embryonic lethality. The major cell type of the fetal portion of the placenta is the trophoblast. Primary mouse placental trophoblast cells are a useful tool for studying normal and abnormal placental development, and unlike cell lines, may be isolated and used to study trophoblast at specific stages of pregnancy. In addition, primary cultures of trophoblast from transgenic mice may be used to study the role of particular genes in placental cells. The protocol presented here is based on the description by Thordarson et al., in which a percoll gradient is used to obtain a relatively pure trophoblast cell population from isolated mouse placentas. It is similar to the more widely used methods for human trophoblast cell isolation. Purity may be assessed by immunocytochemical staining of the isolated cells for cytokeratin 7. Here, the isolated cells are then analyzed using a matrigel invasion assay to assess trophoblast invasiveness in vitro. The invaded cells are analyzed by immunocytochemistry and stained for counting., (Copyright © 2012 Journal of Visualized Experiments)
- Published
- 2012
- Full Text
- View/download PDF
50. Preeclampsia: multiple approaches for a multifactorial disease.
- Author
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Pennington KA, Schlitt JM, Jackson DL, Schulz LC, and Schust DJ
- Subjects
- Animals, Biomedical Research, Clinical Trials as Topic, Disease Models, Animal, Female, Humans, Pre-Eclampsia diagnosis, Pre-Eclampsia physiopathology, Pregnancy, Translational Research, Biomedical, Pre-Eclampsia etiology, Pre-Eclampsia pathology
- Abstract
Preeclampsia is a pregnancy-specific disorder characterized by hypertension and excess protein excretion in the urine. It is an important cause of maternal and fetal morbidity and mortality worldwide. The disease is almost exclusive to humans and delivery of the pregnancy continues to be the only effective treatment. The disorder is probably multifactorial, although most cases of preeclampsia are characterized by abnormal maternal uterine vascular remodeling by fetally derived placental trophoblast cells. Numerous in vitro and animal models have been used to study aspects of preeclampsia, the most common being models of placental oxygen dysregulation, abnormal trophoblast invasion, inappropriate maternal vascular damage and anomalous maternal-fetal immune interactions. Investigations into the pathophysiology and treatment of preeclampsia continue to move the field forward, albeit at a frustratingly slow pace. There remains a pressing need for novel approaches, new disease models and innovative investigators to effectively tackle this complex and devastating disorder.
- Published
- 2012
- Full Text
- View/download PDF
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