133 results on '"Hölter SM"'
Search Results
2. MiR-34a deficiency accelerates medulloblastoma formation in vivo
- Author
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Thor, T, Künkele, A, Pajtler, Kw, Wefers, Ak, Stephan, H, Mestdagh, P, Heukamp, L, Hartmann, W, Vandesompele, J, Sadowski, N, Becker, L, Garrett, L, Hölter, Sm, Horsch, M, Calzada Wack, J, Klein Rodewald, T, Racz, I, Zimmer, A, Beckers, J, Neff, F, Klopstock, T, DE ANTONELLIS, PASQUALINO, ZOLLO, MASSIMO, Wurst, W, Fuchs, H, Gailus Durner, V, Schüller, U, de Angelis, Mh, Eggert, A, Schramm, A, Schulte, J.h., Thor, T, Künkele, A, Pajtler, Kw, Wefers, Ak, Stephan, H, Mestdagh, P, Heukamp, L, Hartmann, W, Vandesompele, J, Sadowski, N, Becker, L, Garrett, L, Hölter, Sm, Horsch, M, Calzada Wack, J, Klein Rodewald, T, Racz, I, Zimmer, A, Beckers, J, Neff, F, Klopstock, T, DE ANTONELLIS, Pasqualino, Zollo, Massimo, Wurst, W, Fuchs, H, Gailus Durner, V, Schüller, U, de Angelis, Mh, Eggert, A, Schramm, A, and Schulte, J. h.
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MIRN34 microRNA, human ,Medizin ,Mice, Transgenic ,metabolism [Cerebellar Neoplasms] ,ND2:SmoA1 ,MIRN34 microRNA, mouse ,Gene Knockout Techniques ,Mice ,metabolism [MicroRNAs] ,metabolism [Medulloblastoma] ,Cell Line, Tumor ,Cerebellum ,MYCN ,Animals ,Humans ,genetics [Cerebellar Neoplasms] ,ddc:610 ,genetics [MicroRNAs] ,Cerebellar Neoplasms ,neoplasms ,Cell Proliferation ,SmoA1 [miR-34a ,ND2] ,genetics [Medulloblastoma] ,pediatric brain tumors ,pathology [Medulloblastoma] ,metabolism [Cerebellum] ,pathology [Cerebellar Neoplasms] ,Gene Expression Regulation, Neoplastic ,MicroRNAs ,Phenotype ,miR-34a ,Medulloblastoma ,Signal Transduction - Abstract
International Journal of Cancer Volume 136, Issue 10, 15 May 2015, Pages 2293-2303 MiR-34a deficiency accelerates medulloblastoma formation in vivo (Article) Thor, T.abc , Künkele, A.a, Pajtler, K.W.a, Wefers, A.K.d, Stephan, H.a, Mestdagh, P.e, Heukamp, L.f, Hartmann, W.g, Vandesompele, J.e, Sadowski, N.a, Becker, L.hi, Garrett, L.hj, Hölter, S.M.hj, Horsch, M.hk, Calzada-Wack, J.hl, Klein-Rodewald, T.hl, Racz, I.hm, Zimmer, A.hm, Beckers, J.hkn, Neff, F.hl, Klopstock, T.i, Antonellis, P.D.op, Zollo, M.op, Wurst, W.hjqrs, Fuchs, H.hk, Gailus-Durner, V.hk, Schüller, U.d, De Angelis, M.H.hkn, Eggert, A.t, Schramm, A.a, Schulte, J.H.abcuv a Department of Pediatric Oncology and Hematology, University Children's Hospital Essen, Hufelandstr. 55, Essen, Germany b German Cancer Consortium (DKTK), Heidelberg, Germany c German Cancer Research Center (DKFZ), Heidelberg, Germany View additional affiliations View references (61) Abstract Previous studies have evaluated the role of miRNAs in cancer initiation and progression. MiR-34a was found to be downregulated in several tumors, including medulloblastomas. Here we employed targeted transgenesis to analyze the function of miR-34a in vivo. We generated mice with a constitutive deletion of the miR-34a gene. These mice were devoid of mir-34a expression in all analyzed tissues, but were viable and fertile. A comprehensive standardized phenotypic analysis including more than 300 single parameters revealed no apparent phenotype. Analysis of miR-34a expression in human medulloblastomas and medulloblastoma cell lines revealed significantly lower levels than in normal human cerebellum. Re-expression of miR-34a in human medulloblastoma cells reduced cell viability and proliferation, induced apoptosis and downregulated the miR-34a target genes, MYCN and SIRT1. Activation of the Shh pathway by targeting SmoA1 transgene overexpression causes medulloblastoma in mice, which is dependent on the presence and upregulation of Mycn. Analysis of miR-34a in medulloblastomas derived from ND2:SmoA1(tg) mice revealed significant suppression of miR-34a compared to normal cerebellum. Tumor incidence was significantly increased and tumor formation was significantly accelerated in mice transgenic for SmoA1 and lacking miR-34a. Interestingly, Mycn and Sirt1 were strongly expressed in medulloblastomas derived from these mice. We here demonstrate that miR-34a is dispensable for normal development, but that its loss accelerates medulloblastomagenesis. Strategies aiming to re-express miR-34a in tumors could, therefore, represent an efficient therapeutic option.
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- 2014
3. ABCB4-defiziente Mäuse zeigen depressives und ängstliches Verhalten
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Hochrath, K, primary, Hölter, SM, additional, Garrett, L, additional, Gailus-Durner, V, additional, Fuchs, H, additional, Hrabě de Angelis, M, additional, Wurst, W, additional, Stokes, CS, additional, and Lammert, F, additional
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- 2014
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4. Die Deletion des Glucosetransporters GLUT8 in Mäusen vermindert die Spermienmotilität und steigert die körperlichen Aktivität
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Gawlik, V, primary, Kluth, O, additional, Schmidt, S, additional, Scheepers, A, additional, Wennemuth, G, additional, Hölter, SM, additional, Augustin, R, additional, Moser, M, additional, Behrens, M, additional, Joost, HG, additional, and Schürmann, A, additional
- Published
- 2009
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5. Creatine improves health and survival in mice
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Haack, T, primary, Bender, A, additional, Beckers, J, additional, Hölter, SM, additional, Ruthsatz, T, additional, Vogt-Weisenhorn, D, additional, Becker, L, additional, Genius, J, additional, Rujescu, D, additional, Irmler, M, additional, Mijalski, T, additional, Mader, M, additional, Quintanilla-Martinez, L, additional, Fuchs, H, additional, Gailus-Durner, V, additional, Hrabé de Angelis, M, additional, Wurst, W, additional, Schmidt, J, additional, Schneider, I, additional, and Klopstock, T, additional
- Published
- 2007
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6. Telomere shortening reduces Alzheimer's disease amyloid pathology in mice.
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Rolyan H, Scheffold A, Heinrich A, Begus-Nahrmann Y, Langkopf BH, Hölter SM, Vogt-Weisenhorn DM, Liss B, Wurst W, Lie DC, Thal DR, Biber K, Rudolph KL, Rolyan, Harshvardhan, Scheffold, Annika, Heinrich, Annette, Begus-Nahrmann, Yvonne, Langkopf, Britta Heike, Hölter, Sabine M, and Vogt-Weisenhorn, Daniela M
- Abstract
Alzheimer's disease is a neurodegenerative disorder of the elderly and advancing age is the major risk factor for Alzheimer's disease development. Telomere shortening represents one of the molecular causes of ageing that limits the proliferative capacity of cells, including neural stem cells. Studies on telomere lengths in patients with Alzheimer's disease have revealed contrary results and the functional role of telomere shortening on brain ageing and Alzheimer's disease is not known. Here, we have investigated the effects of telomere shortening on adult neurogenesis and Alzheimer's disease progression in mice. The study shows that aged telomerase knockout mice with short telomeres (G3Terc-/-) exhibit reduced dentate gyrus neurogenesis and loss of neurons in hippocampus and frontal cortex, associated with short-term memory deficit in comparison to mice with long telomere reserves (Terc+/+). In contrast, telomere shortening improved the spatial learning ability of ageing APP23 transgenic mice, a mouse model for Alzheimer's disease. Telomere shortening was also associated with an activation of microglia in ageing amyloid-free brain. However, in APP23 transgenic mice, telomere shortening reduced both amyloid plaque pathology and reactive microgliosis. Together, these results provide the first experimental evidence that telomere shortening, despite impairing adult neurogenesis and maintenance of post-mitotic neurons, can slow down the progression of amyloid plaque pathology in Alzheimer's disease, possibly involving telomere-dependent effects on microglia activation. [ABSTRACT FROM AUTHOR]
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- 2011
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7. Comparative Phenotyping of Mice Reveals Canonical and Noncanonical Physiological Functions of TRα and TRβ.
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Hönes GS, Geist D, Wenzek C, Pfluger PT, Müller TD, Aguilar-Pimentel JA, Amarie OV, Becker L, Dragano N, Garrett L, Hölter SM, Rathkolb B, Rozman J, Spielmann N, Treise I, Wolf E, Wurst W, Fuchs H, Gailus-Durner V, Hrabe de Angelis M, Führer D, and Moeller LC
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- Animals, Female, Male, Mice, Signal Transduction genetics, Thyroid Hormones metabolism, Mice, Inbred C57BL, Mice, Knockout, Phenotype, Thyroid Hormone Receptors alpha genetics, Thyroid Hormone Receptors alpha metabolism, Thyroid Hormone Receptors beta genetics, Thyroid Hormone Receptors beta metabolism
- Abstract
Thyroid hormone (TH) effects are mediated through TH receptors (TRs), TRα1, TRβ1, and TRβ2. The TRs bind to the DNA and regulate expression of TH target genes (canonical signaling). In addition, they mediate activation of signaling pathways (noncanonical signaling). Whether noncanonical TR action contributes to the spectrum of TH effects is largely unknown. The aim of this study was to attribute physiological effects to the TR isoforms and their canonical and noncanonical signaling. We conducted multiparameter phenotyping in male and female TR knockout mice (TRαKO, TRβKO), mice with disrupted canonical signaling due to mutations in the TR DNA binding domain (TRαGS, TRβGS), and their wild-type littermates. Perturbations in senses, especially hearing (mainly TRβ with a lesser impact of TRα), visual acuity, retinal thickness (TRα and TRβ), and in muscle metabolism (TRα) highlighted the role of canonical TR action. Strikingly, selective abrogation of canonical TR action often had little phenotypic consequence, suggesting that noncanonical TR action sufficed to maintain the wild-type phenotype for specific effects. For instance, macrocytic anemia, reduced retinal vascularization, or increased anxiety-related behavior were only observed in TRαKO but not TRαGS mice. Noncanonical TRα action improved energy utilization and prevented hyperphagia observed in female TRαKO mice. In summary, by examining the phenotypes of TRα and TRβ knockout models alongside their DNA binding-deficient mutants and wild-type counterparts, we could establish that the noncanonical actions of TRα and TRβ play a crucial role in modulating sensory, behavioral, and metabolic functions and, thus, contribute to the spectrum of physiological TH effects., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Endocrine Society. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com. See the journal About page for additional terms.)
- Published
- 2024
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8. A brain-enriched circular RNA controls excitatory neurotransmission and restricts sensitivity to aversive stimuli.
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Giusti SA, Pino NS, Pannunzio C, Ogando MB, Armando NG, Garrett L, Zimprich A, Becker L, Gimeno ML, Lukin J, Merino FL, Pardi MB, Pedroncini O, Di Mauro GC, Durner VG, Fuchs H, de Angelis MH, Patop IL, Turck CW, Deussing JM, Vogt Weisenhorn DM, Jahn O, Kadener S, Hölter SM, Brose N, Giesert F, Wurst W, Marin-Burgin A, and Refojo D
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- Animals, Mice, Mice, Knockout, Neurons metabolism, Neurons physiology, RNA, Circular genetics, Synaptic Transmission, Brain metabolism, Brain physiology
- Abstract
Circular RNAs (circRNAs) are a large class of noncoding RNAs. Despite the identification of thousands of circular transcripts, the biological significance of most of them remains unexplored, partly because of the lack of effective methods for generating loss-of-function animal models. In this study, we focused on circTulp4, an abundant circRNA derived from the Tulp4 gene that is enriched in the brain and synaptic compartments. By creating a circTulp4-deficient mouse model, in which we mutated the splice acceptor site responsible for generating circTulp4 without affecting the linear mRNA or protein levels, we were able to conduct a comprehensive phenotypic analysis. Our results demonstrate that circTulp4 is critical in regulating neuronal and brain physiology, modulating the strength of excitatory neurotransmission and sensitivity to aversive stimuli. This study provides evidence that circRNAs can regulate biologically relevant functions in neurons, with modulatory effects at multiple levels of the phenotype, establishing a proof of principle for the regulatory role of circRNAs in neural processes.
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- 2024
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9. A patient-enriched MEIS1 coding variant causes a restless legs syndrome-like phenotype in mice.
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Leu CL, Lam DD, Salminen AV, Wefers B, Becker L, Garrett L, Rozman J, Wurst W, Hrabě de Angelis M, Hölter SM, Winkelmann J, and Williams RH
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- Animals, Female, Humans, Mice, Age Factors, Disease Models, Animal, Mice, Inbred C57BL, Mutation, Missense genetics, Neoplasm Proteins genetics, Phenotype, Point Mutation genetics, Sleep genetics, Sleep physiology, Homeodomain Proteins genetics, Myeloid Ecotropic Viral Integration Site 1 Protein genetics, Restless Legs Syndrome genetics, Restless Legs Syndrome physiopathology
- Abstract
Restless legs syndrome (RLS) is a neurological disorder characterized by uncomfortable or unpleasant sensations in the legs during rest periods. To relieve these sensations, patients move their legs, causing sleep disruption. While the pathogenesis of RLS has yet to be resolved, there is a strong genetic association with the MEIS1 gene. A missense variant in MEIS1 is enriched sevenfold in people with RLS compared to non-affected individuals. We generated a mouse line carrying this mutation (p.Arg272His/c.815G>A), referred to herein as Meis1R272H/R272H (Meis1 point mutation), to determine whether it would phenotypically resemble RLS. As women are more prone to RLS, driven partly by an increased risk of developing RLS during pregnancy, we focused on female homozygous mice. We evaluated RLS-related outcomes, particularly sensorimotor behavior and sleep, in young and aged mice. Compared to noncarrier littermates, homozygous mice displayed very few differences. Significant hyperactivity occurred before the lights-on (rest) period in aged female mice, reflecting the age-dependent incidence of RLS. Sensory experiments involving tactile feedback (rotarod, wheel running, and hotplate) were only marginally different. Overall, RLS-like phenomena were not recapitulated except for the increased wake activity prior to rest. This is likely due to the focus on young mice. Nevertheless, the Meis1R272H mouse line is a potentially useful RLS model, carrying a clinically relevant variant and showing an age-dependent phenotype., (© The Author(s) 2024. Published by Oxford University Press on behalf of Sleep Research Society. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)
- Published
- 2024
- Full Text
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10. Implication of transcription factor FOXD2 dysfunction in syndromic congenital anomalies of the kidney and urinary tract (CAKUT).
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Riedhammer KM, Nguyen TT, Koşukcu C, Calzada-Wack J, Li Y, Assia Batzir N, Saygılı S, Wimmers V, Kim GJ, Chrysanthou M, Bakey Z, Sofrin-Drucker E, Kraiger M, Sanz-Moreno A, Amarie OV, Rathkolb B, Klein-Rodewald T, Garrett L, Hölter SM, Seisenberger C, Haug S, Schlosser P, Marschall S, Wurst W, Fuchs H, Gailus-Durner V, Wuttke M, Hrabe de Angelis M, Ćomić J, Akgün Doğan Ö, Özlük Y, Taşdemir M, Ağbaş A, Canpolat N, Orenstein N, Çalışkan S, Weber RG, Bergmann C, Jeanpierre C, Saunier S, Lim TY, Hildebrandt F, Alhaddad B, Basel-Salmon L, Borovitz Y, Wu K, Antony D, Matschkal J, Schaaf CW, Renders L, Schmaderer C, Rogg M, Schell C, Meitinger T, Heemann U, Köttgen A, Arnold SJ, Ozaltin F, Schmidts M, and Hoefele J
- Subjects
- Adult, Animals, Humans, Mice, Genome-Wide Association Study, Mice, Knockout, Transcription Factors genetics, Embryonic Structures, Kidney abnormalities, Kidney embryology, Kidney Diseases genetics, Nephrons embryology, Urinary Tract, Urogenital Abnormalities genetics, Vesico-Ureteral Reflux genetics, Forkhead Transcription Factors deficiency, Forkhead Transcription Factors metabolism
- Abstract
Congenital anomalies of the kidney and urinary tract (CAKUT) are the predominant cause for chronic kidney disease below age 30 years. Many monogenic forms have been discovered due to comprehensive genetic testing like exome sequencing. However, disease-causing variants in known disease-associated genes only explain a proportion of cases. Here, we aim to unravel underlying molecular mechanisms of syndromic CAKUT in three unrelated multiplex families with presumed autosomal recessive inheritance. Exome sequencing in the index individuals revealed three different rare homozygous variants in FOXD2, encoding a transcription factor not previously implicated in CAKUT in humans: a frameshift in the Arabic and a missense variant each in the Turkish and the Israeli family with segregation patterns consistent with autosomal recessive inheritance. CRISPR/Cas9-derived Foxd2 knockout mice presented with a bilateral dilated kidney pelvis accompanied by atrophy of the kidney papilla and mandibular, ophthalmologic, and behavioral anomalies, recapitulating the human phenotype. In a complementary approach to study pathomechanisms of FOXD2-dysfunction-mediated developmental kidney defects, we generated CRISPR/Cas9-mediated knockout of Foxd2 in ureteric bud-induced mouse metanephric mesenchyme cells. Transcriptomic analyses revealed enrichment of numerous differentially expressed genes important for kidney/urogenital development, including Pax2 and Wnt4 as well as gene expression changes indicating a shift toward a stromal cell identity. Histology of Foxd2 knockout mouse kidneys confirmed increased fibrosis. Further, genome-wide association studies suggest that FOXD2 could play a role for maintenance of podocyte integrity during adulthood. Thus, our studies help in genetic diagnostics of monogenic CAKUT and in understanding of monogenic and multifactorial kidney diseases., (Copyright © 2023 International Society of Nephrology. Published by Elsevier Inc. All rights reserved.)
- Published
- 2024
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11. A systematic review of the development and application of home cage monitoring in laboratory mice and rats.
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Kahnau P, Mieske P, Wilzopolski J, Kalliokoski O, Mandillo S, Hölter SM, Voikar V, Amfim A, Badurek S, Bartelik A, Caruso A, Čater M, Ey E, Golini E, Jaap A, Hrncic D, Kiryk A, Lang B, Loncarevic-Vasiljkovic N, Meziane H, Radzevičienė A, Rivalan M, Scattoni ML, Torquet N, Trifkovic J, Ulfhake B, Thöne-Reineke C, Diederich K, Lewejohann L, and Hohlbaum K
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- Male, Female, Mice, Animals, Rats, Social Behavior, Heart Rate physiology, Animals, Domestic, Behavior, Animal physiology, Artificial Intelligence
- Abstract
Background: Traditionally, in biomedical animal research, laboratory rodents are individually examined in test apparatuses outside of their home cages at selected time points. However, the outcome of such tests can be influenced by various factors and valuable information may be missed when the animals are only monitored for short periods. These issues can be overcome by longitudinally monitoring mice and rats in their home cages. To shed light on the development of home cage monitoring (HCM) and the current state-of-the-art, a systematic review was carried out on 521 publications retrieved through PubMed and Web of Science., Results: Both the absolute (~ × 26) and relative (~ × 7) number of HCM-related publications increased from 1974 to 2020. There was a clear bias towards males and individually housed animals, but during the past decade (2011-2020), an increasing number of studies used both sexes and group housing. In most studies, animals were kept for short (up to 4 weeks) time periods in the HCM systems; intermediate time periods (4-12 weeks) increased in frequency in the years between 2011 and 2020. Before the 2000s, HCM techniques were predominantly applied for less than 12 h, while 24-h measurements have been more frequent since the 2000s. The systematic review demonstrated that manual monitoring is decreasing in relation to automatic techniques but still relevant. Until (and including) the 1990s, most techniques were applied manually but have been progressively replaced by automation since the 2000s. Independent of the year of publication, the main behavioral parameters measured were locomotor activity, feeding, and social behaviors; the main physiological parameters were heart rate and electrocardiography. External appearance-related parameters were rarely examined in the home cages. Due to technological progress and application of artificial intelligence, more refined and detailed behavioral parameters have been investigated in the home cage more recently., Conclusions: Over the period covered in this study, techniques for HCM of mice and rats have improved considerably. This development is ongoing and further progress as well as validation of HCM systems will extend the applications to allow for continuous, longitudinal, non-invasive monitoring of an increasing range of parameters in group-housed small rodents in their home cages., (© 2023. The Author(s).)
- Published
- 2023
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12. AOX delays the onset of the lethal phenotype in a mouse model of Uqcrh (complex III) disease.
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Jacobs HT, Szibor M, Rathkolb B, da Silva-Buttkus P, Aguilar-Pimentel JA, Amarie OV, Becker L, Calzada-Wack J, Dragano N, Garrett L, Gerlini R, Hölter SM, Klein-Rodewald T, Kraiger M, Leuchtenberger S, Marschall S, Östereicher MA, Pfannes K, Sanz-Moreno A, Seisenberger C, Spielmann N, Stoeger C, Wurst W, Fuchs H, Hrabě de Angelis M, and Gailus-Durner V
- Subjects
- Animals, Mice, Mammals metabolism, Mitochondrial Membranes metabolism, Oxidation-Reduction, Phenotype, Transcription Factors metabolism, Electron Transport Complex III genetics, Electron Transport Complex III metabolism, Mitochondria metabolism
- Abstract
The alternative oxidase, AOX, provides a by-pass of the cytochrome segment of the mitochondrial respiratory chain when the chain is unavailable. AOX is absent from mammals, but AOX from Ciona intestinalis is benign when expressed in mice. Although non-protonmotive, so does not contribute directly to ATP production, it has been shown to modify and in some cases rescue phenotypes of respiratory-chain disease models. Here we studied the effect of C. intestinalis AOX on mice engineered to express a disease-equivalent mutant of Uqcrh, encoding the hinge subunit of mitochondrial respiratory complex III, which results in a complex metabolic phenotype beginning at 4-5 weeks, rapidly progressing to lethality within a further 6-7 weeks. AOX expression delayed the onset of this phenotype by several weeks, but provided no long-term benefit. We discuss the significance of this finding in light of the known and hypothesized effects of AOX on metabolism, redox homeostasis, oxidative stress and cell signaling. Although not a panacea, the ability of AOX to mitigate disease onset and progression means it could be useful in treatment., Competing Interests: Declaration of competing interest The authors declare that they have no conflicts of interest regarding the contents of this article. MSz is a shareholder in a spin-off company founded to develop AOX-based therapeutics., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2023
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13. Monoallelic variation in DHX9, the gene encoding the DExH-box helicase DHX9, underlies neurodevelopment disorders and Charcot-Marie-Tooth disease.
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Calame DG, Guo T, Wang C, Garrett L, Jolly A, Dawood M, Kurolap A, Henig NZ, Fatih JM, Herman I, Du H, Mitani T, Becker L, Rathkolb B, Gerlini R, Seisenberger C, Marschall S, Hunter JV, Gerard A, Heidlebaugh A, Challman T, Spillmann RC, Jhangiani SN, Coban-Akdemir Z, Lalani S, Liu L, Revah-Politi A, Iglesias A, Guzman E, Baugh E, Boddaert N, Rondeau S, Ormieres C, Barcia G, Tan QKG, Thiffault I, Pastinen T, Sheikh K, Biliciler S, Mei D, Melani F, Shashi V, Yaron Y, Steele M, Wakeling E, Østergaard E, Nazaryan-Petersen L, Millan F, Santiago-Sim T, Thevenon J, Bruel AL, Thauvin-Robinet C, Popp D, Platzer K, Gawlinski P, Wiszniewski W, Marafi D, Pehlivan D, Posey JE, Gibbs RA, Gailus-Durner V, Guerrini R, Fuchs H, Hrabě de Angelis M, Hölter SM, Cheung HH, Gu S, and Lupski JR
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- Animals, Humans, Mice, Cell Line, DEAD-box RNA Helicases genetics, Dichlorodiphenyl Dichloroethylene, DNA Helicases, Mammals, Neoplasm Proteins genetics, Charcot-Marie-Tooth Disease genetics, Neurodevelopmental Disorders
- Abstract
DExD/H-box RNA helicases (DDX/DHX) are encoded by a large paralogous gene family; in a subset of these human helicase genes, pathogenic variation causes neurodevelopmental disorder (NDD) traits and cancer. DHX9 encodes a BRCA1-interacting nuclear helicase regulating transcription, R-loops, and homologous recombination and exhibits the highest mutational constraint of all DDX/DHX paralogs but remains unassociated with disease traits in OMIM. Using exome sequencing and family-based rare-variant analyses, we identified 20 individuals with de novo, ultra-rare, heterozygous missense or loss-of-function (LoF) DHX9 variant alleles. Phenotypes ranged from NDDs to the distal symmetric polyneuropathy axonal Charcot-Marie-Tooth disease (CMT2). Quantitative Human Phenotype Ontology (HPO) analysis demonstrated genotype-phenotype correlations with LoF variants causing mild NDD phenotypes and nuclear localization signal (NLS) missense variants causing severe NDD. We investigated DHX9 variant-associated cellular phenotypes in human cell lines. Whereas wild-type DHX9 was restricted to the nucleus, NLS missense variants abnormally accumulated in the cytoplasm. Fibroblasts from an individual with an NLS variant also showed abnormal cytoplasmic DHX9 accumulation. CMT2-associated missense variants caused aberrant nucleolar DHX9 accumulation, a phenomenon previously associated with cellular stress. Two NDD-associated variants, p.Gly411Glu and p.Arg761Gln, altered DHX9 ATPase activity. The severe NDD-associated variant p.Arg141Gln did not affect DHX9 localization but instead increased R-loop levels and double-stranded DNA breaks. Dhx9
-/- mice exhibited hypoactivity in novel environments, tremor, and sensorineural hearing loss. All together, these results establish DHX9 as a critical regulator of mammalian neurodevelopment and neuronal homeostasis., Competing Interests: Declaration of interests J.R.L. has stock ownership in 23andMe, is a paid consultant for Genome International, and is a co-inventor on multiple US and European patents related to molecular diagnostics for inherited neuropathies, eye diseases, genomic disorders, and bacterial genomic fingerprinting. The Department of Molecular and Human Genetics at the Baylor College of Medicine receives revenue from clinical genetic testing conducted at Baylor Genetics (BG) Laboratories. F.M. and T.S.-S. are employees of GeneDx., (Copyright © 2023 American Society of Human Genetics. All rights reserved.)- Published
- 2023
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14. A rationale for considering heart/brain axis control in neuropsychiatric disease.
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Garrett L, Trümbach D, Spielmann N, Wurst W, Fuchs H, Gailus-Durner V, Hrabě de Angelis M, and Hölter SM
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- Humans, Brain, Autonomic Nervous System physiology, Biomarkers, Heart, Heart Failure
- Abstract
Neuropsychiatric diseases (NPD) represent a significant global disease burden necessitating innovative approaches to pathogenic understanding, biomarker identification and therapeutic strategy. Emerging evidence implicates heart/brain axis malfunction in NPD etiology, particularly via the autonomic nervous system (ANS) and brain central autonomic network (CAN) interaction. This heart/brain inter-relationship harbors potentially novel NPD diagnosis and treatment avenues. Nevertheless, the lack of multidisciplinary clinical approaches as well as a limited appreciation of molecular underpinnings has stymied progress. Large-scale preclinical multi-systemic functional data can therefore provide supplementary insight into CAN and ANS interaction. We here present an overview of the heart/brain axis in NPD and establish a unique rationale for utilizing a preclinical cardiovascular disease risk gene set to glean insights into heart/brain axis control in NPD. With a top-down approach focusing on genes influencing electrocardiogram ANS function, we combined hierarchical clustering of corresponding regional CAN expression data and functional enrichment analysis to reveal known and novel molecular insights into CAN and NPD. Through 'support vector machine' inquiries for classification and literature validation, we further pinpointed the top 32 genes highly expressed in CAN brain structures altering both heart rate/heart rate variability (HRV) and behavior. Our observations underscore the potential of HRV/hyperactivity behavior as endophenotypes for multimodal disease biomarker identification to index aberrant executive brain functioning with relevance for NPD. This work heralds the potential of large-scale preclinical functional genetic data for understanding CAN/ANS control and introduces a stepwise design leveraging preclinical data to unearth novel heart/brain axis control genes in NPD., (© 2022. The Author(s).)
- Published
- 2023
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15. Knockout mouse models as a resource for the study of rare diseases.
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da Silva-Buttkus P, Spielmann N, Klein-Rodewald T, Schütt C, Aguilar-Pimentel A, Amarie OV, Becker L, Calzada-Wack J, Garrett L, Gerlini R, Kraiger M, Leuchtenberger S, Östereicher MA, Rathkolb B, Sanz-Moreno A, Stöger C, Hölter SM, Seisenberger C, Marschall S, Fuchs H, Gailus-Durner V, and Hrabě de Angelis M
- Subjects
- Mice, Animals, Humans, Mice, Knockout, Gene Knockout Techniques, Phenotype, Rare Diseases genetics
- Abstract
Rare diseases (RDs) are a challenge for medicine due to their heterogeneous clinical manifestations and low prevalence. There is a lack of specific treatments and only a few hundred of the approximately 7,000 RDs have an approved regime. Rapid technological development in genome sequencing enables the mass identification of potential candidates that in their mutated form could trigger diseases but are often not confirmed to be causal. Knockout (KO) mouse models are essential to understand the causality of genes by allowing highly standardized research into the pathogenesis of diseases. The German Mouse Clinic (GMC) is one of the pioneers in mouse research and successfully uses (preclinical) data obtained from single-gene KO mutants for research into monogenic RDs. As part of the International Mouse Phenotyping Consortium (IMPC) and INFRAFRONTIER, the pan-European consortium for modeling human diseases, the GMC expands these preclinical data toward global collaborative approaches with researchers, clinicians, and patient groups.Here, we highlight proprietary genes that when deleted mimic clinical phenotypes associated with known RD targets (Nacc1, Bach2, Klotho alpha). We focus on recognized RD genes with no pre-existing KO mouse models (Kansl1l, Acsf3, Pcdhgb2, Rabgap1, Cox7a2) which highlight novel phenotypes capable of optimizing clinical diagnosis. In addition, we present genes with intriguing phenotypic data (Zdhhc5, Wsb2) that are not presently associated with known human RDs.This report provides comprehensive evidence for genes that when deleted cause differences in the KO mouse across multiple organs, providing a huge translational potential for further understanding monogenic RDs and their clinical spectrum. Genetic KO studies in mice are valuable to further explore the underlying physiological mechanisms and their overall therapeutic potential., (© 2023. The Author(s).)
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- 2023
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16. Implication of FOXD2 dysfunction in syndromic congenital anomalies of the kidney and urinary tract (CAKUT).
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Riedhammer KM, Nguyen TT, Koşukcu C, Calzada-Wack J, Li Y, Saygılı S, Wimmers V, Kim GJ, Chrysanthou M, Bakey Z, Kraiger M, Sanz-Moreno A, Amarie OV, Rathkolb B, Klein-Rodewald T, Garrett L, Hölter SM, Seisenberger C, Haug S, Marschall S, Wurst W, Fuchs H, Gailus-Durner V, Wuttke M, de Angelis MH, Ćomić J, Doğan ÖA, Özlük Y, Taşdemir M, Ağbaş A, Canpolat N, Ćalışkan S, Weber R, Bergmann C, Jeanpierre C, Saunier S, Lim TY, Hildebrandt F, Alhaddad B, Wu K, Antony D, Matschkal J, Schaaf C, Renders L, Schmaderer C, Meitinger T, Heemann U, Köttgen A, Arnold S, Ozaltin F, Schmidts M, and Hoefele J
- Abstract
Background: Congenital anomalies of the kidney and urinary tract (CAKUT) are the predominant cause for chronic kidney disease below 30 years of age. Many monogenic forms have been discovered mainly due to comprehensive genetic testing like exome sequencing (ES). However, disease-causing variants in known disease-associated genes still only explain a proportion of cases. Aim of this study was to unravel the underlying molecular mechanism of syndromic CAKUT in two multiplex families with presumed autosomal recessive inheritance., Methods and Results: ES in the index individuals revealed two different rare homozygous variants in FOXD2 , a transcription factor not previously implicated in CAKUT in humans: a frameshift in family 1 and a missense variant in family 2 with family segregation patterns consistent with autosomal-recessive inheritance. CRISPR/Cas9-derived Foxd2 knock-out (KO) mice presented with bilateral dilated renal pelvis accompanied by renal papilla atrophy while extrarenal features included mandibular, ophthalmologic, and behavioral anomalies, recapitulating the phenotype of humans with FOXD2 dysfunction. To study the pathomechanism of FOXD2 -dysfunction-mediated developmental renal defects, in a complementary approach, we generated CRISPR/Cas9-mediated KO of Foxd2 in ureteric-bud-induced mouse metanephric mesenchyme cells. Transcriptomic analyses revealed enrichment of numerous differentially expressed genes important in renal/urogenital development, including Pax2 and Wnt4 as well as gene expression changes indicating a cell identity shift towards a stromal cell identity. Histology of Foxd2 KO mouse kidneys confirmed increased fibrosis. Further, GWAS data (genome-wide association studies) suggests that FOXD2 could play a role for maintenance of podocyte integrity during adulthood., Conclusions: In summary, our data implicate that FOXD2 dysfunction is a very rare cause of autosomal recessive syndromic CAKUT and suggest disturbances of the PAX2-WNT4 cell signaling axis contribute to this phenotype., Competing Interests: Competing interests The authors declare no competing interests.
- Published
- 2023
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17. Improving biomedical research by automated behaviour monitoring in the animal home cage - action needed for networking.
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Hölter SM, Wells S, and Voikar V
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- Animals, Humans, Reproducibility of Results, Monitoring, Physiologic, Models, Animal, Behavior, Animal, Biomedical Research
- Abstract
The key goal in biomedical research is a better understanding of disease aetiologies, which ideally results in strategies and recommendations for the prevention of diseases before they arise, and in the development of effective therapies. However, many concerns have been expressed about the reproducibility and the translational validity of preclinical research in animal models to inform clinical trials in humans. It has been proposed that improving internal, external and construct validity of animal studies will lead to improved translatability. Automated behaviour monitoring in the animal's home cage, which allows for longitudinal assessment of individual trajectories over sufficiently long intervals for (chronic) drug treatment or phenotype progression, is a promising solution to these problems.
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- 2023
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18. Recommendations for robust and reproducible preclinical research in personalised medicine.
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Fosse V, Oldoni E, Bietrix F, Budillon A, Daskalopoulos EP, Fratelli M, Gerlach B, Groenen PMA, Hölter SM, Menon JML, Mobasheri A, Osborne N, Ritskes-Hoitinga M, Ryll B, Schmitt E, Ussi A, Andreu AL, and McCormack E
- Subjects
- Humans, Precision Medicine
- Abstract
Background: Personalised medicine is a medical model that aims to provide tailor-made prevention and treatment strategies for defined groups of individuals. The concept brings new challenges to the translational step, both in clinical relevance and validity of models. We have developed a set of recommendations aimed at improving the robustness of preclinical methods in translational research for personalised medicine., Methods: These recommendations have been developed following four main steps: (1) a scoping review of the literature with a gap analysis, (2) working sessions with a wide range of experts in the field, (3) a consensus workshop, and (4) preparation of the final set of recommendations., Results: Despite the progress in developing innovative and complex preclinical model systems, to date there are fundamental deficits in translational methods that prevent the further development of personalised medicine. The literature review highlighted five main gaps, relating to the relevance of experimental models, quality assessment practices, reporting, regulation, and a gap between preclinical and clinical research. We identified five points of focus for the recommendations, based on the consensus reached during the consultation meetings: (1) clinically relevant translational research, (2) robust model development, (3) transparency and education, (4) revised regulation, and (5) interaction with clinical research and patient engagement. Here, we present a set of 15 recommendations aimed at improving the robustness of preclinical methods in translational research for personalised medicine., Conclusions: Appropriate preclinical models should be an integral contributor to interventional clinical trial success rates, and predictive translational models are a fundamental requirement to realise the dream of personalised medicine. The implementation of these guidelines is ambitious, and it is only through the active involvement of all relevant stakeholders in this field that we will be able to make an impact and effectuate a change which will facilitate improved translation of personalised medicine in the future., (© 2023. The Author(s).)
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- 2023
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19. Deep phenotyping and lifetime trajectories reveal limited effects of longevity regulators on the aging process in C57BL/6J mice.
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Xie K, Fuchs H, Scifo E, Liu D, Aziz A, Aguilar-Pimentel JA, Amarie OV, Becker L, da Silva-Buttkus P, Calzada-Wack J, Cho YL, Deng Y, Edwards AC, Garrett L, Georgopoulou C, Gerlini R, Hölter SM, Klein-Rodewald T, Kramer M, Leuchtenberger S, Lountzi D, Mayer-Kuckuk P, Nover LL, Oestereicher MA, Overkott C, Pearson BL, Rathkolb B, Rozman J, Russ J, Schaaf K, Spielmann N, Sanz-Moreno A, Stoeger C, Treise I, Bano D, Busch DH, Graw J, Klingenspor M, Klopstock T, Mock BA, Salomoni P, Schmidt-Weber C, Weiergräber M, Wolf E, Wurst W, Gailus-Durner V, Breteler MMB, Hrabě de Angelis M, and Ehninger D
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- Mice, Animals, Male, Mice, Inbred C57BL, Phenotype, Longevity genetics, Aging physiology
- Abstract
Current concepts regarding the biology of aging are primarily based on studies aimed at identifying factors regulating lifespan. However, lifespan as a sole proxy measure for aging can be of limited value because it may be restricted by specific pathologies. Here, we employ large-scale phenotyping to analyze hundreds of markers in aging male C57BL/6J mice. For each phenotype, we establish lifetime profiles to determine when age-dependent change is first detectable relative to the young adult baseline. We examine key lifespan regulators (putative anti-aging interventions; PAAIs) for a possible countering of aging. Importantly, unlike most previous studies, we include in our study design young treated groups of animals, subjected to PAAIs prior to the onset of detectable age-dependent phenotypic change. Many PAAI effects influence phenotypes long before the onset of detectable age-dependent change, but, importantly, do not alter the rate of phenotypic change. Hence, these PAAIs have limited effects on aging., (© 2022. The Author(s).)
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- 2022
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20. Seizures, ataxia and parvalbumin-expressing interneurons respond to selenium supply in Selenop-deficient mice.
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Schweizer U, Wirth EK, Klopstock T, Hölter SM, Becker L, Moskovitz J, Grune T, Fuchs H, Gailus-Durner V, Hrabe de Angelis M, Köhrle J, and Schomburg L
- Abstract
Mice with constitutive disruption of the Selenop gene have been key to delineate the importance of selenoproteins in neurobiology. However, the phenotype of this mouse model is exquisitely dependent on selenium supply and timing of selenium supplementation. Combining biochemical, histological, and behavioral methods, we tested the hypothesis that parvalbumin-expressing interneurons in the primary somatosensory cortex and hippocampus depend on dietary selenium availability in Selenop
-/- mice. Selenop-deficient mice kept on adequate selenium diet (0.15 mg/kg, i.e. the recommended dietary allowance, RDA) developed ataxia, tremor, and hyperexcitability between the age of 4-5 weeks. Video-electroencephalography demonstrated epileptic seizures in Selenop-/- mice fed the RDA diet, while Selenop± heterozygous mice behaved normally. Both neurological phenotypes, hyperexcitability/seizures and ataxia/dystonia were successfully prevented by selenium supplementation from birth or transgenic expression of human SELENOP under a hepatocyte-specific promoter. Selenium supplementation with 10 μM selenite in the drinking water on top of the RDA diet increased the activity of glutathione peroxidase in the brains of Selenop-/- mice to control levels. The effects of selenium supplementation on the neurological phenotypes were dose- and time-dependent. Selenium supplementation after weaning was apparently too late to prevent ataxia/dystonia, while selenium withdrawal from rescued Selenop-/- mice eventually resulted in ataxia. We conclude that SELENOP expression is essential for preserving interneuron survival under limiting Se supply, while SELENOP appears dispensable under sufficiently high Se status., Competing Interests: Declaration of competing interest L.S. holds shares of selenOmed GmbH, a company involved in Se status assessment. All other authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)- Published
- 2022
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21. A Pathophysiological Intersection of Diabetes and Alzheimer's Disease.
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Čater M and Hölter SM
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- Humans, Obesity metabolism, Alzheimer Disease metabolism, Cognitive Dysfunction complications, Diabetes Mellitus, Type 2 complications, Insulin Resistance physiology
- Abstract
Diabetes is among the most prevalent diseases of the modern world and is strongly linked to an increased risk of numerous neurodegenerative disorders, although the exact pathophysiological mechanisms are not clear yet. Insulin resistance is a serious pathological condition, connecting type 2 diabetes, metabolic syndrome, and obesity. Recently, insulin resistance has been proven to be connected also to cognitive decline and dementias, including the most prevalent form, Alzheimer's disease. The relationship between diabetes and Alzheimer's disease regarding pathophysiology is so significant that it has been proposed that some presentations of the condition could be termed type 3 diabetes.
- Published
- 2022
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22. Parkinson's disease motor symptoms rescue by CRISPRa-reprogramming astrocytes into GABAergic neurons.
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Giehrl-Schwab J, Giesert F, Rauser B, Lao CL, Hembach S, Lefort S, Ibarra IL, Koupourtidou C, Luecken MD, Truong DJ, Fischer-Sternjak J, Masserdotti G, Prakash N, Ninkovic J, Hölter SM, Vogt Weisenhorn DM, Theis FJ, Götz M, and Wurst W
- Subjects
- Animals, Astrocytes, Corpus Striatum, Dopamine, Dopaminergic Neurons, GABAergic Neurons, Mice, Parkinson Disease genetics, Parkinson Disease therapy
- Abstract
Direct reprogramming based on genetic factors resembles a promising strategy to replace lost cells in degenerative diseases such as Parkinson's disease. For this, we developed a knock-in mouse line carrying a dual dCas9 transactivator system (dCAM) allowing the conditional in vivo activation of endogenous genes. To enable a translational application, we additionally established an AAV-based strategy carrying intein-split-dCas9 in combination with activators (AAV-dCAS). Both approaches were successful in reprogramming striatal astrocytes into induced GABAergic neurons confirmed by single-cell transcriptome analysis of reprogrammed neurons in vivo. These GABAergic neurons functionally integrate into striatal circuits, alleviating voluntary motor behavior aspects in a 6-OHDA Parkinson's disease model. Our results suggest a novel intervention strategy beyond the restoration of dopamine levels. Thus, the AAV-dCAS approach might enable an alternative route for clinical therapies of Parkinson's disease., (© 2022 The Authors. Published under the terms of the CC BY 4.0 license.)
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- 2022
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23. Post-synaptic scaffold protein TANC2 in psychiatric and somatic disease risk.
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Garrett L, Da Silva-Buttkus P, Rathkolb B, Gerlini R, Becker L, Sanz-Moreno A, Seisenberger C, Zimprich A, Aguilar-Pimentel A, Amarie OV, Cho YL, Kraiger M, Spielmann N, Calzada-Wack J, Marschall S, Busch D, Schmitt-Weber C, Wolf E, Wurst W, Fuchs H, Gailus-Durner V, Hölter SM, and Hrabě de Angelis M
- Subjects
- Animals, Brain metabolism, Humans, Mice, Neuronal Plasticity, Protein Domains, Neurodevelopmental Disorders genetics, Neurodevelopmental Disorders metabolism, Proteins metabolism
- Abstract
Understanding the shared genetic aetiology of psychiatric and medical comorbidity in neurodevelopmental disorders (NDDs) could improve patient diagnosis, stratification and treatment options. Rare tetratricopeptide repeat, ankyrin repeat and coiled-coil containing 2 (TANC2)-disrupting variants were disease causing in NDD patients. The post-synaptic scaffold protein TANC2 is essential for dendrite formation in synaptic plasticity and plays an unclarified but critical role in development. We here report a novel homozygous-viable Tanc2-disrupted function model in which mutant mice were hyperactive and had impaired sensorimotor gating consistent with NDD patient psychiatric endophenotypes. Yet, a multi-systemic analysis revealed the pleiotropic effects of Tanc2 outside the brain, such as growth failure and hepatocellular damage. This was associated with aberrant liver function including altered hepatocellular metabolism. Integrative analysis indicates that these disrupted Tanc2 systemic effects relate to interaction with Hippo developmental signalling pathway proteins and will increase the risk for comorbid somatic disease. This highlights how NDD gene pleiotropy can augment medical comorbidity susceptibility, underscoring the benefit of holistic NDD patient diagnosis and treatment for which large-scale preclinical functional genomics can provide complementary pleiotropic gene function information., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2022. Published by The Company of Biologists Ltd.)
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- 2022
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24. Characterising a homozygous two-exon deletion in UQCRH: comparing human and mouse phenotypes.
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Vidali S, Gerlini R, Thompson K, Urquhart JE, Meisterknecht J, Aguilar-Pimentel JA, Amarie OV, Becker L, Breen C, Calzada-Wack J, Chhabra NF, Cho YL, da Silva-Buttkus P, Feichtinger RG, Gampe K, Garrett L, Hoefig KP, Hölter SM, Jameson E, Klein-Rodewald T, Leuchtenberger S, Marschall S, Mayer-Kuckuk P, Miller G, Oestereicher MA, Pfannes K, Rathkolb B, Rozman J, Sanders C, Spielmann N, Stoeger C, Szibor M, Treise I, Walter JH, Wurst W, Mayr JA, Fuchs H, Gärtner U, Wittig I, Taylor RW, Newman WG, Prokisch H, Gailus-Durner V, and Hrabě de Angelis M
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- Animals, Electron Transport Complex III, Exons, Homozygote, Humans, Mice, Phenotype, Sequence Deletion, Mitochondrial Diseases genetics
- Abstract
Mitochondrial disorders are clinically and genetically diverse, with isolated complex III (CIII) deficiency being relatively rare. Here, we describe two affected cousins, presenting with recurrent episodes of severe lactic acidosis, hyperammonaemia, hypoglycaemia and encephalopathy. Genetic investigations in both cases identified a homozygous deletion of exons 2 and 3 of UQCRH, which encodes a structural complex III (CIII) subunit. We generated a mouse model with the equivalent homozygous Uqcrh deletion (Uqcrh
-/- ), which also presented with lactic acidosis and hyperammonaemia, but had a more severe, non-episodic phenotype, resulting in failure to thrive and early death. The biochemical phenotypes observed in patient and Uqcrh-/- mouse tissues were remarkably similar, displaying impaired CIII activity, decreased molecular weight of fully assembled holoenzyme and an increase of an unexpected large supercomplex (SXL ), comprising mostly of one complex I (CI) dimer and one CIII dimer. This phenotypic similarity along with lentiviral rescue experiments in patient fibroblasts verifies the pathogenicity of the shared genetic defect, demonstrating that the Uqcrh-/- mouse is a valuable model for future studies of human CIII deficiency., (© 2021 The Authors. Published under the terms of the CC BY 4.0 license.)- Published
- 2021
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25. Introduction to the EQIPD quality system.
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Bespalov A, Bernard R, Gilis A, Gerlach B, Guillén J, Castagné V, Lefevre IA, Ducrey F, Monk L, Bongiovanni S, Altevogt B, Arroyo-Araujo M, Bikovski L, de Bruin N, Castaños-Vélez E, Dityatev A, Emmerich CH, Fares R, Ferland-Beckham C, Froger-Colléaux C, Gailus-Durner V, Hölter SM, Hofmann MC, Kabitzke P, Kas MJ, Kurreck C, Moser P, Pietraszek M, Popik P, Potschka H, Prado Montes de Oca E, Restivo L, Riedel G, Ritskes-Hoitinga M, Samardzic J, Schunn M, Stöger C, Voikar V, Vollert J, Wever KE, Wuyts K, MacLeod MR, Dirnagl U, and Steckler T
- Subjects
- Cooperative Behavior, Data Accuracy, Diffusion of Innovation, Europe, Humans, Interdisciplinary Communication, Quality Control, Quality Improvement, Stakeholder Participation, Biomedical Research standards, Drug Evaluation, Preclinical standards, Research Design standards
- Abstract
While high risk of failure is an inherent part of developing innovative therapies, it can be reduced by adherence to evidence-based rigorous research practices. Supported through the European Union's Innovative Medicines Initiative, the EQIPD consortium has developed a novel preclinical research quality system that can be applied in both public and private sectors and is free for anyone to use. The EQIPD Quality System was designed to be suited to boost innovation by ensuring the generation of robust and reliable preclinical data while being lean, effective and not becoming a burden that could negatively impact the freedom to explore scientific questions. EQIPD defines research quality as the extent to which research data are fit for their intended use. Fitness, in this context, is defined by the stakeholders, who are the scientists directly involved in the research, but also their funders, sponsors, publishers, research tool manufacturers, and collaboration partners such as peers in a multi-site research project. The essence of the EQIPD Quality System is the set of 18 core requirements that can be addressed flexibly, according to user-specific needs and following a user-defined trajectory. The EQIPD Quality System proposes guidance on expectations for quality-related measures, defines criteria for adequate processes (i.e. performance standards) and provides examples of how such measures can be developed and implemented. However, it does not prescribe any pre-determined solutions. EQIPD has also developed tools (for optional use) to support users in implementing the system and assessment services for those research units that successfully implement the quality system and seek formal accreditation. Building upon the feedback from users and continuous improvement, a sustainable EQIPD Quality System will ultimately serve the entire community of scientists conducting non-regulated preclinical research, by helping them generate reliable data that are fit for their intended use., Competing Interests: AB AB is an employee and/or shareholder at PAASP GmbH, PAASP US LLC, Exciva GmbH, Synventa LLC, Ritec Pharma, RB, MA, LB, Nd, EC, AD, RF, VG, SH, MH, MK, MP, PP, EP, LR, GR, MR, JS, MS, CS, VV, JV, KW No competing interests declared, AG AG are employees of Janssen / Johnson & Johnson and shareholders at Johnson & Johnson, BG, CE BG and CE are employees and shareholders at PAASP GmbH. JG JG is an employee of AAALAC International that is an EQIPD Associated Collaborator. VC, CF VC and CFC are employees of Porsolt. IL, FD IAL and FD are employees of Sanofi. LM LM is an employee and shareholder of UCB. SB SB is an employee of Novartis Pharma. BA BA is an employee and shareholder of Pfizer. The views and opinions expressed in this article are those of the individual author and should not be attributed to Pfizer, its directors, officers, employees, affiliates, or any organization with which the author is employed or affiliated. CF AB, BA, NdB, UD, CFB, PK, MK, MM, PM, PP, GR, JS, and TS are members of the Preclinical Data Forum (co-chairs - AB and TS), a network financially and organizationally supported by ECNP and Cohen Veterans Bioscience. PK PK is an employee and shareholder at PAASP US LLC. CK UD and CK receive funding from Volkswagen Foundation, PM PM is owner of Cerbascience Consulting, HP HP has received during the last three years consulting and speaking fees and/or funding for collaborative projects from Bayer, Roche, Zogenix, and Eisai. KW KW is a consultant of Avertim, Brussels, Belgium, support for this contribution was funded by Janssen Pharmaceutica NV. MM MM, UD and TS are members of the Advisory Board at PAASP. MM, UD and TS are members of the ARRIVE guidelines working group. UD UD and CK receive funding from Volkswagen Foundation. MM, UD and TS are members of the Advisory Board at PAASP. MM, UD and TS are members of the ARRIVE guidelines working group. TS MM, UD and TS are members of the Advisory Board at PAASP. MM, UD and TS are members of the ARRIVE guidelines working group. TS is an AAALAC ad-hoc specialist. TS and AG are employees of Janssen / Johnson & Johnson and shareholders at Johnson & Johnson., (© 2021, Bespalov et al.)
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- 2021
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26. Dose-dependent long-term effects of a single radiation event on behaviour and glial cells.
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Ung MC, Garrett L, Dalke C, Leitner V, Dragosa D, Hladik D, Neff F, Wagner F, Zitzelsberger H, Miller G, de Angelis MH, Rößler U, Vogt Weisenhorn D, Wurst W, Graw J, and Hölter SM
- Subjects
- Animals, Dose-Response Relationship, Radiation, Female, Hippocampus radiation effects, Male, Mice, Mice, Inbred C3H, Mice, Inbred C57BL, Motor Activity radiation effects, Whole-Body Irradiation, Xeroderma Pigmentosum Group D Protein genetics, Behavior, Animal radiation effects, Neuroglia radiation effects
- Abstract
Purpose: The increasing use of low-dose ionizing radiation in medicine requires a systematic study of its long-term effects on the brain, behaviour and its possible association with neurodegenerative disease vulnerability. Therefore, we analysed the long-term effects of a single low-dose irradiation exposure at 10 weeks of age compared to medium and higher doses on locomotor, emotion-related and sensorimotor behaviour in mice as well as on hippocampal glial cell populations., Materials and Methods: We determined the influence of radiation dose (0, 0.063, 0.125 or 0.5 Gy), time post-irradiation (4, 12 and 18 months p.i.), sex and genotype (wild type versus mice with Ercc2 DNA repair gene point mutation) on behaviour., Results: The high dose (0.5 Gy) had early-onset adverse effects at 4 months p.i. on sensorimotor recruitment and late-onset negative locomotor effects at 12 and 18 months p.i. Notably, the low dose (0.063 Gy) produced no early effects but subtle late-onset (18 months) protective effects on sensorimotor recruitment and exploratory behaviour. Quantification and morphological characterization of the microglial and the astrocytic cells of the dentate gyrus 24 months p.i. indicated heightened immune activity after high dose irradiation (0.125 and 0.5 Gy) while conversely, low dose (0.063 Gy) induced more neuroprotective features., Conclusion: This is one of the first studies demonstrating such long-term and late-onset effects on brain and behaviour after a single radiation event in adulthood.
- Published
- 2021
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27. Diabetes type 2 risk gene Dusp8 is associated with altered sucrose reward behavior in mice and humans.
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Baumann P, Schriever SC, Kullmann S, Zimprich A, Peter A, Gailus-Durner V, Fuchs H, Hrabe de Angelis M, Wurst W, Tschöp MH, Heni M, Hölter SM, and Pfluger PT
- Subjects
- Animals, Feeding Behavior, Female, Humans, Mice, Motivation, Reward, Diabetes Mellitus, Type 2 genetics, Dual-Specificity Phosphatases genetics, Sucrose
- Abstract
Background: Dusp8 is the first GWAS-identified gene that is predominantly expressed in the brain and has previously been linked with the development of diabetes type 2 in humans. In this study, we unravel how Dusp8 is involved in the regulation of sucrose reward behavior., Methods: Female, chow-fed global Dusp8 WT and KO mice were tested in an observer-independent IntelliCage setup for self-administrative sucrose consumption and preference followed by a progressive ratio task with restricted sucrose access to monitor seeking and motivation behavior. Sixty-three human carriers of the major C and minor T allele of DUSP8 SNP rs2334499 were tested for their perception of food cues by collecting a rating score for sweet versus savory high caloric food., Results: Dusp8 KO mice showed a comparable preference for sucrose, but consumed more sucrose compared to WT mice. In a progressive ratio task, Dusp8 KO females switched to a "trial and error" strategy to find sucrose while control Dusp8 WT mice kept their previously established seeking pattern. Nonetheless, the overall motivation to consume sucrose, and the levels of dopaminergic neurons in the brain areas NAcc and VTA were comparable between genotypes. Diabetes-risk allele carriers of DUSP8 SNP rs2334499 preferred sweet high caloric food compared to the major allele carriers, rating scores for savory food remained comparable between groups., Conclusion: Our data suggest a novel role for Dusp8 in the perception of sweet high caloric food as well as in the control of sucrose consumption and foraging in mice and humans., (© 2020 The Authors. Brain and Behavior published by Wiley Periodicals LLC.)
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- 2021
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28. Posterior subcapsular cataracts are a late effect after acute exposure to 0.5 Gy ionizing radiation in mice.
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Kunze S, Cecil A, Prehn C, Möller G, Ohlmann A, Wildner G, Thurau S, Unger K, Rößler U, Hölter SM, Tapio S, Wagner F, Beyerlein A, Theis F, Zitzelsberger H, Kulka U, Adamski J, Graw J, and Dalke C
- Subjects
- Animals, Mice, Male, Female, Lens, Crystalline radiation effects, Gamma Rays adverse effects, Radiation Dosage, Xeroderma Pigmentosum Group D Protein genetics, Cataract etiology, Cataract pathology, Dose-Response Relationship, Radiation
- Abstract
Purpose: The long-term effect of low and moderate doses of ionizing radiation on the lens is still a matter of debate and needs to be evaluated in more detail., Material and Methods: We conducted a detailed histological analysis of eyes from B6C3F1 mice cohorts after acute gamma irradiation (
60 Co source; 0.063 Gy/min) at young adult age of 10 weeks with doses of 0.063, 0.125, and 0.5 Gy. Sham irradiated (0 Gy) mice were used as controls. To test for genetic susceptibility heterozygous Ercc2 mutant mice were used and compared to wild-type mice of the same strain background. Mice of both sexes were included in all cohorts. Eyes were collected 4 h, 12, 18 and 24 months after irradiation. For a better understanding of the underlying mechanisms, metabolomics analyses were performed in lenses and plasma samples of the same mouse cohorts at 4 and 12 h as well as 12, 18 and 24 months after irradiation. For this purpose, a targeted analysis was chosen., Results: This analysis revealed histological changes particularly in the posterior part of the lens that rarely can be observed by using Scheimpflug imaging, as we reported previously. We detected a significant increase of posterior subcapsular cataracts (PSCs) 18 and 24 months after irradiation with 0.5 Gy (odds ratio 9.3; 95% confidence interval 2.1-41.3) independent of sex and genotype. Doses below 0.5 Gy (i.e. 0.063 and 0.125 Gy) did not significantly increase the frequency of PSCs at any time point. In lenses, we observed a clear effect of sex and aging but not of irradiation or genotype. While metabolomics analyses of plasma from the same mice showed only a sex effect., Conclusions: This article demonstrates a significant radiation-induced increase in the incidence of PSCs, which could not be identified using Scheimpflug imaging as the only diagnostic tool.- Published
- 2021
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29. Physiological relevance of the neuronal isoform of inositol-1,4,5-trisphosphate 3-kinases in mice.
- Author
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Blechner C, Becker L, Fuchs H, Rathkolb B, Prehn C, Adler T, Calzada-Wack J, Garrett L, Gailus-Durner V, Morellini F, Conrad S, Hölter SM, Wolf E, Klopstock T, Adamski J, Busch D, de Angelis MH, Schmeisser MJ, and Windhorst S
- Subjects
- Animals, Caco-2 Cells, Female, Humans, Isoenzymes deficiency, Isoenzymes genetics, Male, Mice, Mice, Knockout, Phosphotransferases (Alcohol Group Acceptor) genetics, Neurons enzymology, Phosphotransferases (Alcohol Group Acceptor) deficiency, Prepulse Inhibition physiology
- Abstract
Inositol-1,4,5-trisphosphate 3-kinase-A (ITPKA) is the neuronal isoform of ITPKs and exhibits both actin bundling and InsP
3 kinase activity. In addition to neurons, ITPKA is ectopically expressed in tumor cells, where its oncogenic activity increases tumor cell malignancy. In order to analyze the physiological relevance of ITPKA, here we performed a broad phenotypic screening of itpka deficient mice. Our data show that among the neurobehavioral tests analyzed, itpka deficient mice reacted faster to a hotplate, prepulse inhibition was impaired and the accelerating rotarod test showed decreased latency of itpka deficient mice to fall. These data indicate that ITPKA is involved in the regulation of nociceptive pathways, sensorimotor gating and motor learning. Analysis of extracerebral functions in control and itpka deficient mice revealed significantly reduced glucose, lactate, and triglyceride plasma concentrations in itpka deficient mice. Based on this finding, expression of ITPKA was analyzed in extracerebral tissues and the highest level was found in the small intestine. However, functional studies on CaCo-2 control and ITPKA depleted cells showed that glucose, as well as triglyceride uptake, were not significantly different between the cell lines. Altogether, these data show that ITPKA exhibits distinct functions in the central nervous system and reveal an involvement of ITPKA in energy metabolism., (Copyright © 2020 Elsevier B.V. All rights reserved.)- Published
- 2020
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30. The rRNA m 6 A methyltransferase METTL5 is involved in pluripotency and developmental programs.
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Ignatova VV, Stolz P, Kaiser S, Gustafsson TH, Lastres PR, Sanz-Moreno A, Cho YL, Amarie OV, Aguilar-Pimentel A, Klein-Rodewald T, Calzada-Wack J, Becker L, Marschall S, Kraiger M, Garrett L, Seisenberger C, Hölter SM, Borland K, Van De Logt E, Jansen PWTC, Baltissen MP, Valenta M, Vermeulen M, Wurst W, Gailus-Durner V, Fuchs H, Hrabe de Angelis M, Rando OJ, Kellner SM, Bultmann S, and Schneider R
- Subjects
- Adenosine analogs & derivatives, Adenosine metabolism, Animals, Mice, Mouse Embryonic Stem Cells cytology, Mouse Embryonic Stem Cells enzymology, Mutation, Pluripotent Stem Cells cytology, Pluripotent Stem Cells metabolism, Protein Biosynthesis genetics, RNA, Ribosomal, 18S metabolism, Cell Differentiation genetics, Gene Expression Regulation, Developmental genetics
- Abstract
Covalent chemical modifications of cellular RNAs directly impact all biological processes. However, our mechanistic understanding of the enzymes catalyzing these modifications, their substrates and biological functions, remains vague. Amongst RNA modifications N
6 -methyladenosine (m6 A) is widespread and found in messenger (mRNA), ribosomal (rRNA), and noncoding RNAs. Here, we undertook a systematic screen to uncover new RNA methyltransferases. We demonstrate that the methyltransferase-like 5 (METTL5) protein catalyzes m6 A in 18S rRNA at position A1832 We report that absence of Mettl5 in mouse embryonic stem cells (mESCs) results in a decrease in global translation rate, spontaneous loss of pluripotency, and compromised differentiation potential. METTL5-deficient mice are born at non-Mendelian rates and develop morphological and behavioral abnormalities. Importantly, mice lacking METTL5 recapitulate symptoms of patients with DNA variants in METTL5 , thereby providing a new mouse disease model. Overall, our biochemical, molecular, and in vivo characterization highlights the importance of m6 A in rRNA in stemness, differentiation, development, and diseases., (© 2020 Ignatova et al.; Published by Cold Spring Harbor Laboratory Press.)- Published
- 2020
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31. Polymorphisms in CRYBB2 encoding βB2-crystallin are associated with antisaccade performance and memory function.
- Author
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Giegling I, Hartmann AM, Genius J, Konte B, Maul S, Straube A, Eggert T, Mulert C, Leicht G, Karch S, Hegerl U, Pogarell O, Hölter SM, Möller HJ, Graw J, and Rujescu D
- Subjects
- Case-Control Studies, Humans, Mutation, Prepulse Inhibition, beta-Crystallin B Chain, Endophenotypes, Sensory Gating
- Abstract
βB2-crystallin (gene symbol: Crybb2/CRYBB2) was first described as a structural protein of the ocular lens before it was detected in various brain regions of the mouse, including the hippocampus and the cerebral cortex. Mutations in the mouse Crybb2 gene lead to alterations of sensorimotor gating measured as prepulse inhibition (PPI) and reduced hippocampal size, combined with an altered number of parvalbumin-positive GABAergic interneurons. Decreased PPI and alterations of parvalbumin-positive interneurons are also endophenotypes that typically occur in schizophrenia. To verify the results found in mice, we genotyped 27 single nucleotide polymorphisms (SNPs) within the CRYBB2 gene and its flanking regions and investigated different schizophrenia typical endophenotypes in a sample of 510 schizophrenia patients and 1322 healthy controls. In the case-control study, no association with schizophrenia was found. However, 3 of the 4 investigated haplotype blocks indicated a decreased CRYBB2 mRNA expression. Two of these blocks were associated with poorer antisaccade task performance and altered working memory-linked functional magnetic resonance imaging signals. For the two haplotypes associated with antisaccade performance, suggestive evidence was found with visual memory and in addition, haplotype block 4 showed a nominally significant association with reduced sensorimotor gating, measured as P50 ratio. These results were not schizophrenia-specific, but could be detected in a combined sample of patients and healthy controls. This is the first study to demonstrate the importance of βB2-crystallin for antisaccade performance and memory function in humans and therefore provides implications for βB2-crystallin function in the human brain.
- Published
- 2020
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32. In-depth phenotyping reveals common and novel disease symptoms in a hemizygous knock-in mouse model (Mut-ko/ki) of mut-type methylmalonic aciduria.
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Lucienne M, Aguilar-Pimentel JA, Amarie OV, Becker L, Calzada-Wack J, da Silva-Buttkus P, Garrett L, Hölter SM, Mayer-Kuckuk P, Rathkolb B, Rozman J, Spielmann N, Treise I, Busch DH, Klopstock T, Schmidt-Weber C, Wolf E, Wurst W, Forny M, Mathis D, Fingerhut R, Froese DS, Gailus-Durner V, Fuchs H, de Angelis MH, and Baumgartner MR
- Subjects
- Animals, Anxiety genetics, Anxiety pathology, Bone Density genetics, Disease Models, Animal, Female, Kidney pathology, Male, Methylmalonic Acid metabolism, Mice, Phenotype, Amino Acid Metabolism, Inborn Errors genetics, Amino Acid Metabolism, Inborn Errors pathology, Methylmalonyl-CoA Mutase deficiency, Methylmalonyl-CoA Mutase genetics
- Abstract
Isolated methylmalonic aciduria (MMAuria) is primarily caused by deficiency of methylmalonyl-CoA mutase (MMUT or MUT). Biochemically, MUT deficiency results in the accumulation of methylmalonic acid (MMA), propionyl-carnitine (C3) and other metabolites. Patients often exhibit lethargy, failure to thrive and metabolic decompensation leading to coma or even death, with kidney and neurological impairment frequently identified in the long-term. Here, we report a hemizygous mouse model which combines a knock-in (ki) missense allele of Mut with a knock-out (ko) allele (Mut-ko/ki mice) that was fed a 51%-protein diet from day 12 of life, constituting a bespoke model of MMAuria. Under this diet, mutant mice developed a pronounced metabolic phenotype characterized by drastically increased blood levels of MMA and C3 compared to their littermate controls (Mut-ki/wt). With this bespoke mouse model, we performed a standardized phenotypic screen to assess the whole-body impairments associated with this strong metabolic condition. We found that Mut-ko/ki mice show common clinical manifestations of MMAuria, including pronounced failure to thrive, indications of mild neurological and kidney dysfunction, and degenerative morphological changes in the liver, along with less well described symptoms such as cardiovascular and hematological abnormalities. The analyses also reveal so far unknown disease characteristics, including low bone mineral density, anxiety-related behaviour and ovarian atrophy. This first phenotypic screening of a MMAuria mouse model confirms its relevance to human disease, reveals new alterations associated with MUT deficiency, and suggests a series of quantifiable readouts that can be used to evaluate potential treatment strategies., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2020
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33. A truncating Aspm allele leads to a complex cognitive phenotype and region-specific reductions in parvalbuminergic neurons.
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Garrett L, Chang YJ, Niedermeier KM, Heermann T, Enard W, Fuchs H, Gailus-Durner V, Angelis MH, Huttner WB, Wurst W, and Hölter SM
- Subjects
- Alleles, Animals, Calmodulin-Binding Proteins, Cognition, Humans, Mice, Mutation, Neurons metabolism, Phenotype, Autism Spectrum Disorder, Nerve Tissue Proteins genetics
- Abstract
Neurodevelopmental disorders are heterogeneous and identifying shared genetic aetiologies and converging signalling pathways affected could improve disease diagnosis and treatment. Truncating mutations of the abnormal spindle-like microcephaly associated (ASPM) gene cause autosomal recessive primary microcephaly (MCPH) in humans. ASPM is a positive regulator of Wnt/β-Catenin signalling and controls symmetric to asymmetric cell division. This process balances neural progenitor proliferation with differentiation during embryogenesis, the malfunction of which could interfere with normal brain development. ASPM mutations may play a role also in other neurodevelopmental disorders, nevertheless, we lack the details of how or to what extent. We therefore assessed neurodevelopmental disease and circuit endophenotypes in mice with a truncating Aspm
1-7 mutation. Aspm1-7 mice exhibited impaired short- and long-term object recognition memory and markedly enhanced place learning in the IntelliCage®. This behaviour pattern is reminiscent of a cognitive phenotype seen in mouse models and patients with a rare form of autism spectrum disorder (ASD) as well as in mouse models of altered Wnt signalling. These alterations were accompanied by ventriculomegaly, corpus callosum dysgenesis and decreased parvalbumin (PV)+ interneuron numbers in the hippocampal Cornu Ammonis (CA) region and thalamic reticular nucleus (TRN). PV+ cell number correlated to object recognition (CA and TRN) and place learning (TRN). This opens the possibility that, as well as causing MCPH, mutant ASPM potentially contributes to other neurodevelopmental disorders such as ASD through altered parvalbuminergic interneuron development affecting cognitive behaviour. These findings provide important information for understanding the genetic overlap and improved treatment of neurodevelopmental disorders associated with ASPM.- Published
- 2020
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34. A comprehensive and comparative phenotypic analysis of the collaborative founder strains identifies new and known phenotypes.
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Kollmus H, Fuchs H, Lengger C, Haselimashhadi H, Bogue MA, Östereicher MA, Horsch M, Adler T, Aguilar-Pimentel JA, Amarie OV, Becker L, Beckers J, Calzada-Wack J, Garrett L, Hans W, Hölter SM, Klein-Rodewald T, Maier H, Mayer-Kuckuk P, Miller G, Moreth K, Neff F, Rathkolb B, Rácz I, Rozman J, Spielmann N, Treise I, Busch D, Graw J, Klopstock T, Wolf E, Wurst W, Yildirim AÖ, Mason J, Torres A, Balling R, Mehaan T, Gailus-Durner V, Schughart K, and Hrabě de Angelis M
- Subjects
- Animals, Collaborative Cross Mice genetics, Databases, Genetic, Female, Genetic Association Studies, Genotype, Male, Mice, Quantitative Trait Loci, Species Specificity, Mice, Inbred Strains genetics, Phenotype
- Abstract
The collaborative cross (CC) is a large panel of mouse-inbred lines derived from eight founder strains (NOD/ShiLtJ, NZO/HILtJ, A/J, C57BL/6J, 129S1/SvImJ, CAST/EiJ, PWK/PhJ, and WSB/EiJ). Here, we performed a comprehensive and comparative phenotyping screening to identify phenotypic differences and similarities between the eight founder strains. In total, more than 300 parameters including allergy, behavior, cardiovascular, clinical blood chemistry, dysmorphology, bone and cartilage, energy metabolism, eye and vision, immunology, lung function, neurology, nociception, and pathology were analyzed; in most traits from sixteen females and sixteen males. We identified over 270 parameters that were significantly different between strains. This study highlights the value of the founder and CC strains for phenotype-genotype associations of many genetic traits that are highly relevant to human diseases. All data described here are publicly available from the mouse phenome database for analyses and downloads.
- Published
- 2020
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35. Dusp8 affects hippocampal size and behavior in mice and humans.
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Baumann P, Schriever SC, Kullmann S, Zimprich A, Feuchtinger A, Amarie O, Peter A, Walch A, Gailus-Durner V, Fuchs H, Hrabě de Angelis M, Wurst W, Tschöp MH, Heni M, Hölter SM, and Pfluger PT
- Subjects
- Adult, Animals, Apoptosis genetics, Apoptosis physiology, Cell Proliferation genetics, Cell Proliferation physiology, Dual-Specificity Phosphatases genetics, Female, Humans, Male, Maze Learning, Mice, Mice, Knockout, Neurogenesis genetics, Neurogenesis physiology, Phosphorylation genetics, Phosphorylation physiology, Young Adult, p38 Mitogen-Activated Protein Kinases genetics, p38 Mitogen-Activated Protein Kinases metabolism, Dual-Specificity Phosphatases metabolism, Hippocampus metabolism
- Abstract
Dual-specificity phosphatase 8 (Dusp8) acts as physiological inhibitor for the MAPKs Jnk, Erk and p38 which are involved in regulating multiple CNS processes. While Dusp8 expression levels are high in limbic areas such as the hippocampus, the functional role of Dusp8 in hippocampus morphology, MAPK-signaling, neurogenesis and apoptosis as well as in behavior are still unclear. It is of particular interest whether human carriers of a DUSP8 allelic variant show similar hippocampal alterations to mice. Addressing these questions using Dusp8 WT and KO mouse littermates, we found that KOs suffered from mildly impaired spatial learning, increased locomotor activity and elevated anxiety. Cell proliferation, apoptosis and p38 and Jnk phosphorylation were unaffected, but phospho-Erk levels were higher in hippocampi of the KOs. Consistent with a decreased hippocampus size in Dusp8 KO mice, we found reduced volumes of the hippocampal subregions subiculum and CA4 in humans carrying the DUSP8 allelic variant SNP rs2334499:C > T. Overall, aberrations in morphology and behavior in Dusp8 KO mice and a decrease in hippocampal volume of SNP rs2334499:C > T carriers point to a novel, translationally relevant role of Dusp8 in hippocampus function that warrants further studies on the role of Dusp8 within the limbic network.
- Published
- 2019
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36. Claudin-12 is not required for blood-brain barrier tight junction function.
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Castro Dias M, Coisne C, Baden P, Enzmann G, Garrett L, Becker L, Hölter SM, Hrabě de Angelis M, Deutsch U, and Engelhardt B
- Subjects
- Animals, Blood-Brain Barrier metabolism, Claudins genetics, Encephalomyelitis, Autoimmune, Experimental metabolism, Endothelial Cells physiology, Female, Gene Knock-In Techniques, Male, Mice, Inbred C57BL, Mice, Transgenic, Tight Junctions metabolism, Blood-Brain Barrier physiology, Claudins physiology, Encephalomyelitis, Autoimmune, Experimental physiopathology, Tight Junctions physiology
- Abstract
Background: The blood-brain barrier (BBB) ensures central nervous system (CNS) homeostasis by strictly controlling the passage of molecules and solutes from the bloodstream into the CNS. Complex and continuous tight junctions (TJs) between brain endothelial cells block uncontrolled paracellular diffusion of molecules across the BBB, with claudin-5 being its dominant TJs protein. However, claudin-5 deficient mice still display ultrastructurally normal TJs, suggesting the contribution of other claudins or tight-junction associated proteins in establishing BBB junctional complexes. Expression of claudin-12 at the BBB has been reported, however the exact function and subcellular localization of this atypical claudin remains unknown., Methods: We created claudin-12-lacZ-knock-in C57BL/6J mice to explore expression of claudin-12 and its role in establishing BBB TJs function during health and neuroinflammation. We furthermore performed a broad standardized phenotypic check-up of the mouse mutant., Results: Making use of the lacZ reporter allele, we found claudin-12 to be broadly expressed in numerous organs. In the CNS, expression of claudin-12 was detected in many cell types with very low expression in brain endothelium. Claudin-12
lacZ/lacZ C57BL/6J mice lacking claudin-12 expression displayed an intact BBB and did not show any signs of BBB dysfunction or aggravated neuroinflammation in an animal model for multiple sclerosis. Determining the precise localization of claudin-12 at the BBB was prohibited by the fact that available anti-claudin-12 antibodies showed comparable detection and staining patterns in tissues from wild-type and claudin-12lacZ/lacZ C57BL/6J mice., Conclusions: Our present study thus shows that claudin-12 is not essential in establishing or maintaining BBB TJs integrity. Claudin-12 is rather expressed in cells that typically lack TJs suggesting that claudin-12 plays a role other than forming classical TJs. At the same time, in depth phenotypic screening of clinically relevant organ functions of claudin-12lacZ/lacZ C57BL/6J mice suggested the involvement of claudin-12 in some neurological but, more prominently, in cardiovascular functions.- Published
- 2019
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37. A mouse model for intellectual disability caused by mutations in the X-linked 2'‑O‑methyltransferase Ftsj1 gene.
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Jensen LR, Garrett L, Hölter SM, Rathkolb B, Rácz I, Adler T, Prehn C, Hans W, Rozman J, Becker L, Aguilar-Pimentel JA, Puk O, Moreth K, Dopatka M, Walther DJ, von Bohlen Und Halbach V, Rath M, Delatycki M, Bert B, Fink H, Blümlein K, Ralser M, Van Dijck A, Kooy F, Stark Z, Müller S, Scherthan H, Gecz J, Wurst W, Wolf E, Zimmer A, Klingenspor M, Graw J, Klopstock T, Busch D, Adamski J, Fuchs H, Gailus-Durner V, de Angelis MH, von Bohlen Und Halbach O, Ropers HH, and Kuss AW
- Subjects
- Animals, Behavior, Animal, Cognition Disorders etiology, Cognition Disorders pathology, Family, Female, Intellectual Disability pathology, Male, Methyltransferases genetics, Methyltransferases metabolism, Mice, Mice, Inbred C57BL, Mice, Knockout, Nociceptive Pain etiology, Nociceptive Pain pathology, Nuclear Proteins metabolism, tRNA Methyltransferases genetics, tRNA Methyltransferases metabolism, Disease Models, Animal, Intellectual Disability etiology, Mental Retardation, X-Linked genetics, Methyltransferases physiology, Mutation, Nuclear Proteins genetics, tRNA Methyltransferases physiology
- Abstract
Mutations in the X chromosomal tRNA 2'‑O‑methyltransferase FTSJ1 cause intellectual disability (ID). Although the gene is ubiquitously expressed affected individuals present no consistent clinical features beyond ID. In order to study the pathological mechanism involved in the aetiology of FTSJ1 deficiency-related cognitive impairment, we generated and characterized an Ftsj1 deficient mouse line based on the gene trapped stem cell line RRD143. Apart from an impaired learning capacity these mice presented with several statistically significantly altered features related to behaviour, pain sensing, bone and energy metabolism, the immune and the hormone system as well as gene expression. These findings show that Ftsj1 deficiency in mammals is not phenotypically restricted to the brain but affects various organ systems. Re-examination of ID patients with FTSJ1 mutations from two previously reported families showed that several features observed in the mouse model were recapitulated in some of the patients. Though the clinical spectrum related to Ftsj1 deficiency in mouse and man is variable, we suggest that an increased pain threshold may be more common in patients with FTSJ1 deficiency. Our findings demonstrate novel roles for Ftsj1 in maintaining proper cellular and tissue functions in a mammalian organism., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2019
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38. Low catalytic activity is insufficient to induce disease pathology in triosephosphate isomerase deficiency.
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Segal J, Mülleder M, Krüger A, Adler T, Scholze-Wittler M, Becker L, Calzada-Wack J, Garrett L, Hölter SM, Rathkolb B, Rozman J, Racz I, Fischer R, Busch DH, Neff F, Klingenspor M, Klopstock T, Grüning NM, Michel S, Lukaszewska-McGreal B, Voigt I, Hartmann L, Timmermann B, Lehrach H, Wolf E, Wurst W, Gailus-Durner V, Fuchs H, H de Angelis M, Schrewe H, Yuneva M, and Ralser M
- Subjects
- Anemia, Hemolytic, Congenital Nonspherocytic enzymology, Animals, Behavior, Animal, Carbohydrate Metabolism, Inborn Errors enzymology, Disease Models, Animal, Enzyme Stability, Female, Humans, Male, Mice, Mice, Inbred C57BL, Mutation, Protein Multimerization, Anemia, Hemolytic, Congenital Nonspherocytic pathology, Carbohydrate Metabolism, Inborn Errors pathology, Catalytic Domain genetics, Triose-Phosphate Isomerase deficiency, Triose-Phosphate Isomerase genetics
- Abstract
Triosephosphate isomerase (TPI) deficiency is a fatal genetic disorder characterized by hemolytic anemia and neurological dysfunction. Although the enzyme defect in TPI was discovered in the 1960s, the exact etiology of the disease is still debated. Some aspects indicate the disease could be caused by insufficient enzyme activity, whereas other observations indicate it could be a protein misfolding disease with tissue-specific differences in TPI activity. We generated a mouse model in which exchange of a conserved catalytic amino acid residue (isoleucine to valine, Ile170Val) reduces TPI specific activity without affecting the stability of the protein dimer. TPI
Ile170Val/Ile170Val mice exhibit an approximately 85% reduction in TPI activity consistently across all examined tissues, which is a stronger average, but more consistent, activity decline than observed in patients or symptomatic mouse models that carry structural defect mutant alleles. While monitoring protein expression levels revealed no evidence for protein instability, metabolite quantification indicated that glycolysis is affected by the active site mutation. TPIIle170Val/Ile170Val mice develop normally and show none of the disease symptoms associated with TPI deficiency. Therefore, without the stability defect that affects TPI activity in a tissue-specific manner, a strong decline in TPI catalytic activity is not sufficient to explain the pathological onset of TPI deficiency., (© 2019 The Authors. Journal of Inherited Metabolic Disease published by John Wiley & Sons Ltd on behalf of SSIEM.)- Published
- 2019
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39. Crybb2 Mutations Consistently Affect Schizophrenia Endophenotypes in Mice.
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Heermann T, Garrett L, Wurst W, Fuchs H, Gailus-Durner V, Hrabě de Angelis M, Graw J, and Hölter SM
- Subjects
- Alleles, Amino Acid Sequence, Animals, Calcium metabolism, Cell Count, Exons genetics, Mice, Inbred C3H, Mice, Inbred C57BL, Parvalbumins metabolism, Prepulse Inhibition, Sensory Gating, beta-Crystallin B Chain chemistry, Endophenotypes metabolism, Mutation genetics, Schizophrenia genetics, beta-Crystallin B Chain genetics
- Abstract
As part of the βγ-superfamily, βB2-crystallin (CRYBB2) is an ocular structural protein in the lens, and mutation of the corresponding gene can cause cataracts. CRYBB2 also is expressed in non-lens tissue such as the adult mouse brain and is associated with neuropsychiatric disorders such as schizophrenia. Nevertheless, the robustness of this association as well as how CRYBB2 may contribute to disease-relevant phenotypes is unknown. To add further clarity to this issue, we performed a comprehensive analysis of behavioral and neurohistological alterations in mice with an allelic series of mutations in the C-terminal end of the Crybb2 gene. Behavioral phenotyping of these three βB2-mutant lines Crybb2
O377 , Crybb2Philly , and Crybb2Aey2 included assessment of exploratory activity and anxiety-related behavior in the open field, sensorimotor gating measured by prepulse inhibition (PPI) of the acoustic startle reflex, cognitive performance measured by social discrimination, and spontaneous alternation in the Y-maze. In each mutant line, we also quantified the number of parvalbumin-positive (PV+) GABAergic interneurons in selected brain regions that express CRYBB2. While there were allele-specific differences in individual behaviors and affected brain areas, all three mutant lines exhibited consistent alterations in PPI that paralleled alterations in the PV+ cell number in the thalamic reticular nucleus (TRN). The direction of the PPI change mirrored that of the TRN PV+ cell number thereby suggesting a role for TRN PV+ cell number in modulating PPI. Moreover, as both altered PPI and PV+ cell number are schizophrenia-associated endophenotypes, our result implicates mutated Crybb2 in the development of this neuropsychiatric disorder.- Published
- 2019
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40. Understanding gene functions and disease mechanisms: Phenotyping pipelines in the German Mouse Clinic.
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Fuchs H, Aguilar-Pimentel JA, Amarie OV, Becker L, Calzada-Wack J, Cho YL, Garrett L, Hölter SM, Irmler M, Kistler M, Kraiger M, Mayer-Kuckuk P, Moreth K, Rathkolb B, Rozman J, da Silva Buttkus P, Treise I, Zimprich A, Gampe K, Hutterer C, Stöger C, Leuchtenberger S, Maier H, Miller M, Scheideler A, Wu M, Beckers J, Bekeredjian R, Brielmeier M, Busch DH, Klingenspor M, Klopstock T, Ollert M, Schmidt-Weber C, Stöger T, Wolf E, Wurst W, Yildirim AÖ, Zimmer A, Gailus-Durner V, and Hrabě de Angelis M
- Subjects
- Animals, Humans, Disease Models, Animal, Mice, Transgenic, Phenotype
- Abstract
Since decades, model organisms have provided an important approach for understanding the mechanistic basis of human diseases. The German Mouse Clinic (GMC) was the first phenotyping facility that established a collaboration-based platform for phenotype characterization of mouse lines. In order to address individual projects by a tailor-made phenotyping strategy, the GMC advanced in developing a series of pipelines with tests for the analysis of specific disease areas. For a general broad analysis, there is a screening pipeline that covers the key parameters for the most relevant disease areas. For hypothesis-driven phenotypic analyses, there are thirteen additional pipelines with focus on neurological and behavioral disorders, metabolic dysfunction, respiratory system malfunctions, immune-system disorders and imaging techniques. In this article, we give an overview of the pipelines and describe the scientific rationale behind the different test combinations., (Copyright © 2017 Elsevier B.V. All rights reserved.)
- Published
- 2018
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41. Male offspring born to mildly ZIKV-infected mice are at risk of developing neurocognitive disorders in adulthood.
- Author
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Stanelle-Bertram S, Walendy-Gnirß K, Speiseder T, Thiele S, Asante IA, Dreier C, Kouassi NM, Preuß A, Pilnitz-Stolze G, Müller U, Thanisch S, Richter M, Scharrenberg R, Kraus V, Dörk R, Schau L, Herder V, Gerhauser I, Pfankuche VM, Käufer C, Waltl I, Moraes T, Sellau J, Hoenow S, Schmidt-Chanasit J, Jansen S, Schattling B, Ittrich H, Bartsch U, Renné T, Bartenschlager R, Arck P, Cadar D, Friese MA, Vapalahti O, Lotter H, Benites S, Rolling L, Gabriel M, Baumgärtner W, Morellini F, Hölter SM, Amarie O, Fuchs H, Hrabe de Angelis M, Löscher W, Calderon de Anda F, and Gabriel G
- Subjects
- Animals, Animals, Newborn, Brain pathology, Disease Models, Animal, Female, Humans, Infectious Disease Transmission, Vertical, Learning Disabilities etiology, Male, Neurocognitive Disorders pathology, Neurocognitive Disorders physiopathology, Placental Insufficiency, Pregnancy, Sex Factors, Testosterone blood, Zika Virus Infection transmission, Neurocognitive Disorders etiology, Pregnancy Complications, Infectious, Zika Virus, Zika Virus Infection complications
- Abstract
Congenital Zika virus (ZIKV) syndrome may cause fetal microcephaly in ~1% of affected newborns. Here, we investigate whether the majority of clinically inapparent newborns might suffer from long-term health impairments not readily visible at birth. Infection of immunocompetent pregnant mice with high-dose ZIKV caused severe offspring phenotypes, such as fetal death, as expected. By contrast, low-dose (LD) maternal ZIKV infection resulted in reduced fetal birth weight but no other obvious phenotypes. Male offspring born to LD ZIKV-infected mothers had increased testosterone (TST) levels and were less likely to survive in utero infection compared to their female littermates. Males also presented an increased number of immature neurons in apical and basal hippocampal dendrites, while female offspring had immature neurons in basal dendrites only. Moreover, male offspring with high but not very high (storm) TST levels were more likely to suffer from learning and memory impairments compared to females. Future studies are required to understand the impact of TST on neuropathological and neurocognitive impairments in later life. In summary, increased sex-specific vigilance is required in countries with high ZIKV prevalence, where impaired neurodevelopment may be camouflaged by a healthy appearance at birth.
- Published
- 2018
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42. Fgf9 Y162C Mutation Alters Information Processing and Social Memory in Mice.
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Garrett L, Becker L, Rozman J, Puk O, Stoeger T, Yildirim AÖ, Bohla A, Eickelberg O, Hans W, Prehn C, Adamski J, Klopstock T, Rácz I, Zimmer A, Klingenspor M, Fuchs H, Gailus-Durner V, Wurst W, Hrabě de Angelis M, Graw J, and Hölter SM
- Subjects
- Analysis of Variance, Anesthesia, Animals, Anxiety genetics, Anxiety physiopathology, Corticosterone metabolism, Discrimination, Psychological, Female, Hippocampus pathology, Hippocampus physiopathology, Male, Mice, Mice, Mutant Strains, Motor Activity, Neurogenesis, Olfactory Bulb pathology, Olfactory Bulb physiopathology, Reflex, Startle, Sex Characteristics, Fibroblast Growth Factor 9 genetics, Memory, Mutation genetics, Social Behavior
- Abstract
In neuropsychiatric diseases, such as major depression and anxiety, pathogenic vulnerability is partially dictated by a genetic predisposition. The search continues to define this genetic susceptibility and establish new genetic elements as potential therapeutic targets. The fibroblast growth factors (FGFs) could be interesting in this regard. This family of signaling molecules plays important roles in development while also functioning within the adult. This includes effects on aspects of brain function such as neurogenesis and synapse formation. Of this family, Fgf9 is expressed in the adult brain, but its functional role is less well defined. In this study, we examined the role of Fgf9 in different brain functions by analyzing the behavior of Fgf9
Y162C mutant mice, an Fgf9 allele without the confounding systemic effects of other Fgf9 genetic models. Here, we show that this mutation caused altered locomotor and exploratory reactivity to novel, mildly stressful environments. In addition, mutants showed heightened acoustic startle reactivity as well as impaired social discrimination memory. Notably, there was a substantial decrease in the level of adult olfactory bulb neurogenesis with no difference in hippocampal neurogenesis. Collectively, our findings indicate a role for the Fgf9Y162C mutation in information processing and perception of aversive situations as well as in social memory. Thus, genetic alterations in Fgf9 could increase vulnerability to developing neuropsychiatric disease, and we propose the Fgf9Y162C mutant mice as a valuable tool to study the predictive etiological aspects.- Published
- 2018
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43. A paternal methyl donor-rich diet altered cognitive and neural functions in offspring mice.
- Author
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Ryan DP, Henzel KS, Pearson BL, Siwek ME, Papazoglou A, Guo L, Paesler K, Yu M, Müller R, Xie K, Schröder S, Becker L, Garrett L, Hölter SM, Neff F, Rácz I, Rathkolb B, Rozman J, Ehninger G, Klingenspor M, Klopstock T, Wolf E, Wurst W, Zimmer A, Fuchs H, Gailus-Durner V, Hrabě de Angelis M, Sidiropoulou K, Weiergräber M, Zhou Y, and Ehninger D
- Subjects
- Animals, DNA Methylation, Diet, Epigenesis, Genetic, Fathers, Folic Acid metabolism, Hippocampus metabolism, Large-Conductance Calcium-Activated Potassium Channel beta Subunits, Learning drug effects, Male, Memory drug effects, Methionine metabolism, Methionine Adenosyltransferase, Methylation, Mice, Mice, Inbred C57BL, Neurons physiology, Paternal Inheritance genetics, Promoter Regions, Genetic, Cognition physiology, Dietary Supplements adverse effects, Paternal Inheritance physiology
- Abstract
Dietary intake of methyl donors, such as folic acid and methionine, shows considerable intra-individual variation in human populations. While it is recognized that maternal departures from the optimum of dietary methyl donor intake can increase the risk for mental health issues and neurological disorders in offspring, it has not been explored whether paternal dietary methyl donor intake influences behavioral and cognitive functions in the next generation. Here, we report that elevated paternal dietary methyl donor intake in a mouse model, transiently applied prior to mating, resulted in offspring animals (methyl donor-rich diet (MD) F1 mice) with deficits in hippocampus-dependent learning and memory, impaired hippocampal synaptic plasticity and reduced hippocampal theta oscillations. Gene expression analyses revealed altered expression of the methionine adenosyltransferase Mat2a and BK channel subunit Kcnmb2, which was associated with changes in Kcnmb2 promoter methylation in MD F1 mice. Hippocampal overexpression of Kcnmb2 in MD F1 mice ameliorated altered spatial learning and memory, supporting a role of this BK channel subunit in the MD F1 behavioral phenotype. Behavioral and gene expression changes did not extend into the F2 offspring generation. Together, our data indicate that paternal dietary factors influence cognitive and neural functions in the offspring generation.
- Published
- 2018
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44. Lifetime study in mice after acute low-dose ionizing radiation: a multifactorial study with special focus on cataract risk.
- Author
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Dalke C, Neff F, Bains SK, Bright S, Lord D, Reitmeir P, Rößler U, Samaga D, Unger K, Braselmann H, Wagner F, Greiter M, Gomolka M, Hornhardt S, Kunze S, Kempf SJ, Garrett L, Hölter SM, Wurst W, Rosemann M, Azimzadeh O, Tapio S, Aubele M, Theis F, Hoeschen C, Slijepcevic P, Kadhim M, Atkinson M, Zitzelsberger H, Kulka U, and Graw J
- Subjects
- Animals, Cataract etiology, Chromosome Aberrations radiation effects, Dose-Response Relationship, Radiation, Female, Kaplan-Meier Estimate, Male, Mice, Radiation Injuries, Experimental etiology, Radiation Protection, Risk Assessment, Telomere radiation effects, Time Factors, Cataract genetics, Radiation Injuries, Experimental genetics
- Abstract
Because of the increasing application of ionizing radiation in medicine, quantitative data on effects of low-dose radiation are needed to optimize radiation protection, particularly with respect to cataract development. Using mice as mammalian animal model, we applied a single dose of 0, 0.063, 0.125 and 0.5 Gy at 10 weeks of age, determined lens opacities for up to 2 years and compared it with overall survival, cytogenetic alterations and cancer development. The highest dose was significantly associated with increased body weight and reduced survival rate. Chromosomal aberrations in bone marrow cells showed a dose-dependent increase 12 months after irradiation. Pathological screening indicated a dose-dependent risk for several types of tumors. Scheimpflug imaging of the lens revealed a significant dose-dependent effect of 1% of lens opacity. Comparison of different biological end points demonstrated long-term effects of low-dose irradiation for several biological end points.
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- 2018
- Full Text
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45. Laboratory mouse housing conditions can be improved using common environmental enrichment without compromising data.
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André V, Gau C, Scheideler A, Aguilar-Pimentel JA, Amarie OV, Becker L, Garrett L, Hans W, Hölter SM, Janik D, Moreth K, Neff F, Östereicher M, Racz I, Rathkolb B, Rozman J, Bekeredjian R, Graw J, Klingenspor M, Klopstock T, Ollert M, Schmidt-Weber C, Wolf E, Wurst W, Gailus-Durner V, Brielmeier M, Fuchs H, and Hrabé de Angelis M
- Subjects
- Animal Welfare economics, Animals, Energy Metabolism physiology, Female, Heart Function Tests methods, Male, Maze Learning physiology, Mice, Mice, Inbred C57BL, Mice, Inbred DBA, Nociception physiology, Animal Welfare ethics, Environment, Controlled, Nesting Behavior physiology
- Abstract
Animal welfare requires the adequate housing of animals to ensure health and well-being. The application of environmental enrichment is a way to improve the well-being of laboratory animals. However, it is important to know whether these enrichment items can be incorporated in experimental mouse husbandry without creating a divide between past and future experimental results. Previous small-scale studies have been inconsistent throughout the literature, and it is not yet completely understood whether and how enrichment might endanger comparability of results of scientific experiments. Here, we measured the effect on means and variability of 164 physiological parameters in 3 conditions: with nesting material with or without a shelter, comparing these 2 conditions to a "barren" regime without any enrichments. We studied a total of 360 mice from each of 2 mouse strains (C57BL/6NTac and DBA/2NCrl) and both sexes for each of the 3 conditions. Our study indicates that enrichment affects the mean values of some of the 164 parameters with no consistent effects on variability. However, the influence of enrichment appears negligible compared to the effects of other influencing factors. Therefore, nesting material and shelters may be used to improve animal welfare without impairment of experimental outcome or loss of comparability to previous data collected under barren housing conditions.
- Published
- 2018
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46. Analysis of locomotor behavior in the German Mouse Clinic.
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Zimprich A, Östereicher MA, Becker L, Dirscherl P, Ernst L, Fuchs H, Gailus-Durner V, Garrett L, Giesert F, Glasl L, Hummel A, Rozman J, de Angelis MH, Vogt-Weisenhorn D, Wurst W, and Hölter SM
- Subjects
- Animals, Female, Genotype, Male, Mice, Mice, Transgenic, Phenotype, Principal Component Analysis, Behavior, Animal physiology, Behavioral Research methods, Gait Analysis methods, Locomotion physiology
- Abstract
Background: Generation and phenotyping of mutant mouse models continues to increase along with the search for the most efficient phenotyping tests. Here we asked if a combination of different locomotor tests is necessary for comprehensive locomotor phenotyping, or if a large data set from an automated gait analysis with the CatWalk system would suffice., New Method: First we endeavored to meaningfully reduce the large CatWalk data set by Principal Component Analysis (PCA) to decide on the most relevant parameters. We analyzed the influence of sex, body weight, genetic background and age. Then a combination of different locomotor tests was analyzed to investigate the possibility of redundancy between tests., Result: The extracted 10 components describe 80% of the total variance in the CatWalk, characterizing different aspects of gait. With these, effects of CatWalk version, sex, body weight, age and genetic background were detected. In addition, the PCA on a combination of locomotor tests suggests that these are independent without significant redundancy in their locomotor measures., Comparison With Existing Methods: The PCA has permitted the refinement of the highly dimensional CatWalk (and other tests) data set for the extraction of individual component scores and subsequent analysis., Conclusion: The outcome of the PCA suggests the possibility to focus on measures of the front and hind paws, and one measure of coordination in future experiments to detect phenotypic differences. Furthermore, although the CatWalk is sensitive for detecting locomotor phenotypes pertaining to gait, it is necessary to include other tests for comprehensive locomotor phenotyping., (Copyright © 2017 Elsevier B.V. All rights reserved.)
- Published
- 2018
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47. RNase H2 Loss in Murine Astrocytes Results in Cellular Defects Reminiscent of Nucleic Acid-Mediated Autoinflammation.
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Bartsch K, Damme M, Regen T, Becker L, Garrett L, Hölter SM, Knittler K, Borowski C, Waisman A, Glatzel M, Fuchs H, Gailus-Durner V, Hrabe de Angelis M, and Rabe B
- Subjects
- Animals, Autoimmune Diseases metabolism, Autoimmune Diseases pathology, Autoimmune Diseases of the Nervous System etiology, Autoimmune Diseases of the Nervous System metabolism, Autoimmune Diseases of the Nervous System pathology, Biomarkers, Brain metabolism, Cells, Cultured, Disease Models, Animal, Encephalomyelitis, Autoimmune, Experimental, Female, Fluorescent Antibody Technique, Immunohistochemistry, Inflammation metabolism, Inflammation pathology, Male, Mice, Mice, Knockout, Nervous System Malformations etiology, Nervous System Malformations metabolism, Nervous System Malformations pathology, Phenotype, Astrocytes immunology, Astrocytes metabolism, Autoimmune Diseases etiology, Inflammation etiology, Nucleic Acids immunology, Ribonuclease H deficiency
- Abstract
Aicardi-Goutières syndrome (AGS) is a rare early onset childhood encephalopathy caused by persistent neuroinflammation of autoimmune origin. AGS is a genetic disorder and >50% of affected individuals bear hypomorphic mutations in ribonuclease H2 (RNase H2). All available RNase H2 mouse models so far fail to mimic the prominent CNS involvement seen in AGS. To establish a mouse model recapitulating the human disease, we deleted RNase H2 specifically in the brain, the most severely affected organ in AGS. Although RNase H2
ΔGFAP mice lacked the nuclease in astrocytes and a majority of neurons, no disease signs were apparent in these animals. We additionally confirmed these results in a second, neuron-specific RNase H2 knockout mouse line. However, when astrocytes were isolated from brains of RNase H2ΔGFAP mice and cultured under mitogenic conditions, they showed signs of DNA damage and premature senescence. Enhanced expression of interferon-stimulated genes (ISGs) represents the most reliable AGS biomarker. Importantly, primary RNase H2ΔGFAP astrocytes displayed significantly increased ISG transcript levels, which we failed to detect in in vivo in brains of RNase H2ΔGFAP mice. Isolated astrocytes primed by DNA damage, including RNase H2-deficiency, exhibited a heightened innate immune response when exposed to bacterial or viral antigens. Taken together, we established a valid cellular AGS model that utilizes the very cell type responsible for disease pathology, the astrocyte, and phenocopies major molecular defects observed in AGS patient cells.- Published
- 2018
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48. Assessing Sociability, Social Memory, and Pup Retrieval in Mice.
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Zimprich A, Niessing J, Cohen L, Garrett L, Einicke J, Sperling B, Schmidt MV, and Hölter SM
- Subjects
- Animals, Maternal Behavior, Memory, Mice physiology, Models, Animal, Social Behavior
- Abstract
Adaptive social behavior is important in mammals, both for the well-being of the individual and for the thriving of the species. Dysfunctions in social behavior occur in many neurodevelopmental and psychiatric diseases, and research into the genetic components of disease-relevant social deficits can open up new avenues for understanding the underlying biological mechanisms and therapeutic interventions. Genetically modified mouse models are particularly useful in this respect, and robust experimental protocols are needed to reliably assess relevant social behavior phenotypes. Here we describe in detail three protocols to quantitatively measure sociability, one of the most frequently investigated social behavior phenotypes in mice, using a three-chamber sociability test. These protocols can be extended to also assess social memory. In addition, we provide a detailed protocol on pup retrieval, which is a particularly robust maternal behavior amenable to various scientific questions. © 2017 by John Wiley & Sons, Inc., (Copyright © 2017 John Wiley & Sons, Inc.)
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- 2017
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49. Serum Response Factor (SRF) Ablation Interferes with Acute Stress-Associated Immediate and Long-Term Coping Mechanisms.
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Zimprich A, Mroz G, Meyer Zu Reckendorf C, Anastasiadou S, Förstner P, Garrett L, Hölter SM, Becker L, Rozman J, Prehn C, Rathkolb B, Moreth K, Wurst W, Klopstock T, Klingenspor M, Adamski J, Wolf E, Bekeredjian R, Fuchs H, Gailus-Durner V, de Angelis MH, and Knöll B
- Subjects
- Animals, Female, Male, Mice, Mice, Inbred C57BL, Mice, Transgenic, Prosencephalon pathology, Serum Response Factor genetics, Stress, Psychological genetics, Stress, Psychological pathology, Time Factors, Adaptation, Psychological physiology, Genes, Immediate-Early physiology, Prosencephalon metabolism, Serum Response Factor deficiency, Stress, Psychological metabolism
- Abstract
Stress experience modulates behavior, metabolism, and energy expenditure of organisms. One molecular hallmark of an acute stress response is a rapid induction of immediate early genes (IEGs) such as c-Fos and Egr family members. IEG transcription in neurons is mediated by the neuronal activity-driven gene regulator serum response factor (SRF). We show a first role of SRF in immediate and long-lasting acute restraint stress (AS) responses. For this, we employed a standardized mouse phenotyping protocol at the German Mouse Clinic (GMC) including behavioral, metabolic, and cardiologic tests as well as gene expression profiling to analyze the consequences of forebrain-specific SRF deletion in mice exposed to AS. Adult mice with an SRF deletion in glutamatergic neurons (Srf;
CaMKIIa-CreERT2 ) showed hyperactivity, decreased anxiety, and impaired working memory. In response to restraint AS, instant stress reactivity including locomotor behavior and corticosterone induction was impaired in Srf mutant mice. Interestingly, even several weeks after previous AS exposure, SRF-deficient mice showed long-lasting AS-associated changes including altered locomotion, metabolism, energy expenditure, and cardiovascular changes. This suggests a requirement of SRF for mediating long-term stress coping mechanisms in wild-type mice. SRF ablation decreased AS-mediated IEG induction and activity of the actin severing protein cofilin. In summary, our data suggest an SRF function in immediate AS reactions and long-term post-stress-associated coping mechanisms.- Published
- 2017
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50. The pathogenic LRRK2 R1441C mutation induces specific deficits modeling the prodromal phase of Parkinson's disease in the mouse.
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Giesert F, Glasl L, Zimprich A, Ernst L, Piccoli G, Stautner C, Zerle J, Hölter SM, Vogt Weisenhorn DM, and Wurst W
- Subjects
- Animals, Arginine genetics, Cysteine genetics, Disease Models, Animal, Exploratory Behavior physiology, Gait genetics, Genotype, Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 metabolism, Mice, Mice, Transgenic, Motor Activity genetics, Parkinson Disease pathology, Recognition, Psychology physiology, Smell genetics, Swimming psychology, Synaptophysin metabolism, Synaptotagmin I metabolism, Tyrosine 3-Monooxygenase metabolism, Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 genetics, Parkinson Disease genetics, Parkinson Disease physiopathology, Point Mutation genetics, Prodromal Symptoms
- Abstract
The aim of the present study was to further explore the in vivo function of the Leucine-rich repeat kinase 2 (LRRK2)-gene, which is mutated in certain familial forms of Parkinson's disease (PD). We generated a mouse model harboring the disease-associated point mutation R1441C in the GTPase domain of the endogenous murine LRRK2 gene (LRRK2 R1441C line) and performed a comprehensive analysis of these animals throughout lifespan in comparison with an existing knockdown line of LRRK2 (LRRK2 knockdown line). Animals of both lines do not exhibit severe motor dysfunction or pathological signs of neurodegeneration neither at young nor old age. However, at old age the homozygous LRRK2 R1441C animals exhibit clear phenotypes related to the prodromal phase of PD such as impairments in fine motor tasks, gait, and olfaction. These phenotypes are only marginally observable in the LRRK2 knockdown animals, possibly due to activation of compensatory mechanisms as suggested by in vitro studies of synaptic transmission. Thus, at the organismal level the LRRK2 R1441C mutation does not emerge as a loss of function of the protein, but induces mutation specific deficits. Furthermore, judged by the phenotypes presented, the LRRK2-R1441C knock-in line is a valid preclinical model for the prodromal phase of PD., (Copyright © 2017. Published by Elsevier Inc.)
- Published
- 2017
- Full Text
- View/download PDF
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