Search

As a part of the routine neritic zooplankton collection program in Obhur Creek (central Red Sea, Saudi Arabia), specimens of a pontellid calanoid copepod, Calanopia media Gurney, 1927, were observed and studied. Since the original description was rather brief and drawings limited, especially of mouthparts and legs, which were not illustrated and described, the species is here fully redescribed. Red Sea specimens showed considerable variation in the female genital compound somite, the right caudal ramus and leg 5, as well as in the presence of a medial knob ventrally on the male right prosomal corner. DNA sequences of mtCOI of different specimens did not show any significant differences and supported their identity as one species. Calanopia media exhibited clear diel vertical migration, with high densities of 106 and 150 ind. m-3 during sunset (6:00 pm; UTC+3) and midnight (12:00 am; UTC+3) collections, respectively. However, this species was not observed in other zooplankton collections from the surface to 20 m depth, at 6:00 am and 12:00 pm (UTC+3). [ABSTRACT FROM AUTHOR]

The Andaman Sea adjoining the Indian Territory is biologically rich in terms of marine diversity. However, inadequate surveys during the post-independence era have resulted in large lacunae in data on the species composition in these waters. In this paper, we report the first observation of two species of dendrobranchiate prawns, Metapenaeopsis difficilis Crosnier, 1991 and Haliporus taprobanensis Alcock and Anderson, 1899, in the Indian seas, supplemented with notes on their morphological diagnoses, comparison with closely related species and zoogeographical distribution. Crustacean specimens were collected using trawl nets and a naturalists' dredge on board the Fisheries Oceanography Research Vessel Sagar Sampada in the deeper offshore region between 124 and 850 m depth in the Andaman Sea during the period 2015–2017. Metapenaeopsis difficilis is distinguished by an oval-shaped thelycal plate and a low, unarmed bead-like plate on the thoracic sternum between the third pereiopods. Metapenaeopsis difficilis is previously reported from the Philippines, Indonesia, Coral Sea, New Caledonia, Marquesas Islands, and Wallis and Futuna Islands at depths between 21 and 440 m, thereby indicating its westward range extension. Haliporus taprobanensis is distinguished by a tough integument, presence of a postero-dorsal spine on the fourth pleonal somite, and smooth dorsum of the first four pleonal somites. Haliporus taprobanensis is previously reported from South Africa, Madagascar, off Sri Lanka, Indonesia, Philippines and northern Australia at depths between 300 and 1650 m. [ABSTRACT FROM AUTHOR]

The problem of vertebrate head metamerism was first posed more than 200 years ago. This issue gave rise to two approaches: segmentalism, which relies on the model of the typical head segment, and antisegmentalism, which rejects the existence of cephalic segments altogether. The truth, probably, lies somewhere in the middle. Data on the development of the head mesoderm, cranial neural crest, and cranial nerves lead to the conclusion that there are five metameres in the vertebrate head: premandibular, mandibular, hyoid, 1st metahyoid, and 2nd metahyoid. According to the hypothesis of F. Edgeworth, this number of head metameres is primary; the head spreading back occurred as a result of the polymerization of visceral arches inside the nervus vagus metamere which had originally included only the 2nd metahyoid arch. According to the hypothesis of C. Gegenbaur, initially, the visceral arches were more numerous in this area and the branchial nerves merged into a single nervus vagus system only secondarily. The Cambrian paleontology may assist in resolving this controversy. In any case, vertebrate head metamerism is primarily heteronomous (as interpreted by P.P. Ivanov): the premandibular and mandibular metameres are morphologically atypical and lie beyond the Hox gene expression zone. Developmental biology shows that the bodies of other metameric animals are arranged on the basis of similar principles. [ABSTRACT FROM AUTHOR]

The calanoid copepod Lucicutia hulsemannae Markhaseva & Ferrari, 2005 has, until now, only been recorded from the eastern tropical Pacific Ocean, i.e., in its description as a new species. The present study now reports L. hulsemannae for the first time from the southern basin of the South China Sea (7°59.57′N 113°0.1′E), at depths of 500-800 m. The main morphological characteristics of females of this species are: (i) Cephalosome with a pair of strong, triangular, lateral protrusions; (ii) plug of genital double-somite conical in lateral view, with swollen base; (iii) second urosomal somite significantly wider and shorter than third; (iv) inner marginal seta of second exopodal segment of fifth pereiopod (P5) thin towards its tip. This finding considerably expands the reported global distribution of L. hulsemannae , at the same time updating the checklist of copepods for Chinese waters. [ABSTRACT FROM AUTHOR]

Zebrafish are a valuable vertebrate model in which to study development and characterize genes involved in cystic kidney disease. Zebrafish embryos and larvae are transparent, allowing non-invasive imaging during their rapid development, which takes place over the first 72 hours post fertilisation. Gene-specific knockdown of nephronophthisis-associated genes leads to ciliary phenotypes which can be assessed in various developmental structures. Here we describe in detail the methods used for imaging cilia within Kupffer’s vesicle to assess nephronophthisis and related ciliopathy phenotypes.

Somites form during embryonic development and give rise to unique cell and tissue types, such as skeletal muscles and bones and cartilage of the vertebrae. Using somitogenesis-stage human embryos, we performed transcriptomic profiling of human presomitic mesoderm as well as nascent and developed somites. In addition to conserved pathways such as WNT-β-catenin, we also identified BMP and transforming growth factor β (TGF-β) signaling as major regulators unique to human somitogenesis. This information enabled us to develop an efficient protocol to derive somite cells in vitro from human pluripotent stem cells (hPSCs). Importantly, the in-vitro-differentiating cells progressively expressed markers of the distinct developmental stages that are known to occur during in vivo somitogenesis. Furthermore, when subjected to lineage-specific differentiation conditions, the hPSC-derived somite cells were multipotent in generating somite derivatives, including skeletal myocytes, osteocytes, and chondrocytes. This work improves our understanding of human somitogenesis and may enhance our ability to treat diseases affecting somite derivatives.

The embryological origin of the trapezius and sternocleidomastoid muscles has been debated for over a century. To shed light on this issue, the present anatomical study was performed. Five fresh frozen human cadavers, three males and two females, were used for this study. Samples from each specimen's trapezius and sternocleidomastoid were fixed in 10% formalin and placed in paraffin blocks. As Paired like homeodomain 2 (Pitx2) and T-box factor 1(Tbx1) have been implicated in the region and muscle type regulation, we performed Tbx1 and Pitx2 Immunohistochemistry (IHC) on these muscle tissue samples to identify the origin of the trapezius and sternocleidomastoid muscles. We have used the latest version of QuPath, v0.4.3, software to quantify the Tbx and Pitx2 staining. For the sternocleidomastoid muscle, for evaluated samples, the average amount of positively stained Tbx1 and Pitx2 was 25% (range 16%-30%) and 18% (range 12%-23%), respectively. For the trapezius muscles, for evaluated samples, the average amount of positively stained Tbx1 and Pitx2 parts of the samples was 17% (range 15%-20%) and 15% (14%-17%), respectively. Our anatomical findings suggest dual origins of both the trapezius and sternocleidomastoid muscles. Additionally, as neither Pitx2 nor Tbx1 made up all the staining observed for each muscle, other contributions to these structures are likely. Future studies with larger samples are now necessary to confirm these findings., (© 2023 American Association of Clinical Anatomists.)

Paraxial mesoderm in the early embryo is segmented into epithelial blocks called somites that establish the metameric organization of the vertebrate body plan. Somites are sequentially formed from head to tail in a rhythmic manner controlled by an oscillating gene regulatory network known as the segmentation clock. We know very little about this important process during human development due to limited access to human embryos and ethical concerns. To bypass these difficulties, model systems derived from human pluripotent stem cells have been established. Here, we detail three protocols modeling different aspects of human paraxial mesoderm development in vitro: a 2D cell monolayer system recapitulating dynamics of the human segmentation clock, a 3D organoid system called "somitoid" supporting the simultaneous formation of somite-like structures, and another organoid system called "segmentoid" reconstituting in vivo-like hallmarks of somitogenesis. Together, these complementary model systems provide an excellent platform to decode somitogenesis and advance human developmental biology., (© 2023. Springer Science+Business Media, LLC.)

In vertebrates, head and trunk muscles develop from different mesodermal populations and are regulated by distinct genetic networks. Neck muscles at the head-trunk interface remain poorly defined due to their complex morphogenesis and dual mesodermal origins. Here, we use genetically modified mice to establish a 3D model that integrates regulatory genes, cell populations and morphogenetic events that define this transition zone. We show that the evolutionary conserved cucullaris-derived muscles originate from posterior cardiopharyngeal mesoderm, not lateral plate mesoderm, and we define new boundaries for neural crest and mesodermal contributions to neck connective tissue. Furthermore, lineage studies and functional analysis of Tbx1- and Pax3-null mice reveal a unique developmental program for somitic neck muscles that is distinct from that of somitic trunk muscles. Our findings unveil the embryological and developmental requirements underlying tetrapod neck myogenesis and provide a blueprint to investigate how muscle subsets are selectively affected in some human myopathies.

Somites are epithelial segments of the paraxial mesoderm. Shortly after their formation, the epithelial somites undergo extensive cellular rearrangements and form specific somite compartments, including the sclerotome and the myotome, which give rise to the axial skeleton and to striated musculature, respectively. The dynamics of somite development varies along the body axis, but most research has focused on somite development at thoracolumbar levels. The development of tail somites has not yet been thoroughly characterized, even though vertebrate tail development has been intensely studied recently with respect to the termination of segmentation and the limitation of body length in evolution. Here, we provide a detailed description of the somites in the avian tail from the beginning of tail formation at HH‐stage 20 to the onset of degeneration of tail segments at HH‐stage 27. We characterize the formation of somite compartment formation in the tail region with respect to morphology and the expression patterns of the sclerotomal marker gene paired‐box gene 1 (Pax1) and the myotomal marker genes MyoD and myogenic factor 5 (Myf5). Our study gives insight into the development of the very last segments formed in the avian embryo, and provides a basis for further research on the development of tail somite derivatives such as tail vertebrae, pygostyle and tail musculature. [ABSTRACT FROM AUTHOR]

Background: Skeletal muscle in the trunk derives from the somites, paired segments of paraxial mesoderm. Whereas axial musculature develops within the somite, appendicular muscle develops following migration of muscle precursors into lateral plate mesoderm. The development of muscles bridging axial and appendicular systems appears mixed. Results: We examine development of three migratory muscle precursor‐derived muscles in zebrafish: the sternohyoideus (SH), pectoral fin (PF), and posterior hypaxial (PHM) muscles. We show there is an anterior to posterior gradient to the developmental gene expression and maturation of these three muscles. SH muscle precursors exhibit a long delay between migration and differentiation, PF muscle precursors exhibit a moderate delay in differentiation, and PHM muscle precursors show virtually no delay between migration and differentiation. Using lineage tracing, we show that lateral plate contribution to the PHM muscle is minor, unlike its known extensive contribution to the PF muscle and absence in the ventral extension of axial musculature. Conclusions: We propose that PHM development is intermediate between a migratory muscle mode and an axial muscle mode of development, wherein the PHM differentiates after a very short migration of its precursors and becomes more anterior primarily by elongation of differentiated muscle fibers. Key Findings: The posterior hypaxial muscle straddles the axial and appendicular systems in teleosts.The posterior hypaxial muscle is neither a simple ventral extension of axial myotome nor a classic MMP‐derived muscle like the pectoral fin muscles.The development of the posterior hypaxial muscle is intermediate to axial and appendicular muscle development. [ABSTRACT FROM AUTHOR]

The two somite compartments, dorso-lateral dermomyotome and medio-ventral sclerotome are major vertebrate novelties, but little is known about their evolutionary origin. We determined that sclerotome cells in Xenopus come from lateral somitic frontier (LSF) by lineage tracing, ablation experiments and histological analysis. We identified Twist1 as marker of migrating sclerotome progenitors in two amphibians, Xenopus and axolotl. From these results, three conclusions can be drawn. First, LSF is made up of multipotent somitic cells (MSCs) since LSF gives rise to sclerotome but also to dermomytome as already shown in Xenopus. Second, the basic scheme of somite compartmentalization is conserved from cephalochordates to anamniotes since in both cases, lateral cells envelop dorsally and ventrally the ancestral myotome, suggesting that lateral MSCs should already exist in cephalochordates. Third, the transition from anamniote to amniote vertebrates is characterized by extension of the MSCs domain to the entire somite at the expense of ancestral myotome since amniote somite is a naive tissue that subdivides into sclerotome and dermomyotome. Like neural crest pluripotent cells, MSCs are at the origin of major vertebrate novelties, namely hypaxial region of the somite, dermomyotome and sclerotome compartments. Hence, change in MSCs properties and location is involved in somite evolution. • Sclerotome progenitors originate from lateral somitic frontier. • Lateral somitic frontier is made of multipotent somitic cells. • Somite compartmentalization is conserved between cephalochordates and anamniotes. • Multipotent somitic cells domain is expended to entire somite in amniote vertebrates. [ABSTRACT FROM AUTHOR]

qRT-PCR requires reliable internal control genes stably expressed in different samples and experimental conditions. The stability of reference genes is rarely tested experimentally, especially in developing tissues given the singularity of these samples. Here we evaluated the suitability of a set of reference genes (Actb , Gapdh , Tbp , Pgk1 and Sdha) using samples from early mouse embryo tissues that are widely used in research (somites, prosencephalon and heart) at different developmental stages. The comparative ΔCq method and five software packages (NormFinder, geNorm, BestKeeper, DataAssist and RefFinder) were used to rank the most stable genes while GenEx and GeNorm programs determined the optimal total number of reference genes for a reliable normalization. The ranking of most reliable reference genes was different for each tissue evaluated: (1) in somite from embryos with 16–18 somite pairs stage, the combination of Pgk1 and Actb provided the best normalization and Actb also presented high stability levels at an earlier developmental stage; (2) Gapdh is the most stable gene in prosencephalon in the two developmental stages tested; and (3) in heart samples, Sdha , Gapdh and Actb were the best combination for qPCR normalization. The analysis of these three tissues simultaneously indicated the combination of Gapdh , Actb and Tbp as the most reliable internal control. This study highlights the importance of appropriate reference genes according to the cell type and/or tissue of interest. The data here described can be applied in future research using mouse embryos as a model for mammalian development. • Standard reference genes (Actb / Gapdh / Tbp/Pgk1 / Sdha) have variable expression stability in early mouse embryo. • In somites the combination of Pgk1 and Actb provided the best normalization. • Gapdh was the most stable gene in prosencephalon. • In the developing heart Sdha , Gapdh and Actb were the top ranked reference genes. • When these 3 tissues were merged the combination of Gapdh , Actb and Tbp was the best normalization. [ABSTRACT FROM AUTHOR]

Muscle stem cells hold a great therapeutic potential in regenerating damaged muscles. However, the in vivo behavior of muscle stem cells during muscle growth and regeneration is still poorly understood. Using zebrafish as a model, we describe the in vivo dynamics and function of embryonic muscle progenitor cells (MPCs) in the dermomyotome. These cells are located in a superficial layer external to muscle fibers and express many extracellular matrix (ECM) genes, including collagen type 1 a2 (col1a2). Utilizing a new col1a2 transgenic line, we show that col1a2+ MPCs display a ramified morphology with dynamic cellular processes. Cell lineage tracing demonstrates that col1a2+ MPCs contribute to new myofibers in normal muscle growth and also during muscle regeneration. A combination of live imaging and single cell clonal analysis reveals a highly choreographed process of muscle regeneration. Activated col1a2+ MPCs change from the quiescent ramified morphology to a polarized and elongated morphology, generating daughter cells that fuse with existing myofibers. Partial depletion of col1a2+ MPCs severely compromises muscle regeneration. Our work provides a dynamic view of embryonic muscle progenitor cells during zebrafish muscle growth and regeneration. [ABSTRACT FROM AUTHOR]

The genus Xanthocalanus inhabits the intermediate and deep waters of the world ocean. The present study records and describes the first female individual of Xanthocalanus agilis Giesbrecht, 1893, collected in the South China Sea (13°0.21′N 113°0.21′E) at depths of 500-800 m. This species was identified by the following morphological characteristics: (i) rostrum with 2 slender filaments; (ii) posterolateral corners of fifth thoracic somite protuberant and triangular, reaching the posterior margin of genital somite; (iii) first segment of fifth pereiopod (P5) with a list of inner marginal spinules, distal segment short with 3 robust terminal spines. This finding expands the known global and vertical distribution of X. agilis. [ABSTRACT FROM AUTHOR]

Somitogenesis is a critical process during vertebrate development that establishes the segmented body plan and gives rise to the vertebra, skeletal muscles, and dermis. While segmentation clock and wave front mechanisms have been elucidated to control the size and time of somite formation, regulation of the segmentation process that physically separates somites is not understood in detail. Here, we identified a cytoskeletal player, Cdc42 effector protein 3 (Cdc42ep3, CEP3) that is required for somite segmentation in Xenopus embryos. CEP3 is specifically expressed in somite tissue during somite segmentation. Loss-of-function experiments showed that CEP3 is not required for the specification of paraxial mesoderm, nor the differentiation of muscle cells, but is required for the segmentation process. Live imaging analysis further revealed that CEP3 is required for cell shape changes and alignment during somitogenesis. When CEP3 was knocked down, somitic cells did not elongate efficiently along the mediolateral axis and failed to undertake the 90° rotation. As a result, cells remained in a continuous sheet without an apparent segmentation cleft. CEP3 likely interacts with Cdc42 during this process, and both increased and decreased Cdc42 activity led to defective somite segmentation. Segmentation defects caused by Cdc42 knockdown can be partially rescued by the overexpression of CEP3. Conversely, loss of CEP3 resulted in the maintenance of high levels of Cdc42 activity at the cell membrane, which is normally reduced during and after somite segmentation. These results suggest that there is a feedback regulation between Cdc42 and CEP3 during somite segmentation and the activity of Cdc42 needs to be fine-tuned to control the coordinated cell shape changes and movement required for somite segmentation. [ABSTRACT FROM AUTHOR]

A model of somite formation in vertebrates is presented in which a biochemical wave selects alternating regions of vertebra and somite production. It does so by imposing expression of a relevant gene in the underlying DNA in the form of a relaxation oscillation. [ABSTRACT FROM AUTHOR]

Lead (Pb) is one of the most abundant toxic metals in the environment that can cause a variety of harmful effects. During embryonic development of vertebrates, somites are temporary organs that give rise to skeletal muscle, cartilage, tendon, endothelial cells, and dermis. In this study, we investigated the effects of lead on the development of somites and their derivatives in chick embryos under in vitro conditions. For this propose, fertilized eggs of Gallus gallus domesticus were incubated until they reached the stage of 15–20 somites. The somites and notochord were isolated and treated with different concentrations of lead acetate (500, 1000, 2000, and 4000 ng ml−1) for 72 h. Our results indicated that high concentrations of lead reduced the nucleus diameter, reduced the synthesis of collagen, inhibited the formation of the cartilage matrix in somite cells, and disturbed the formation and order of myotubes. In conclusion, the results of the current study for the first time indicated the disturbing effects of lead on the development of somites in the chick embryo. Our results revealed that lead disturbed the development of somites in the chick embryo, which suggested that at high concentrations it can cause a serious mortal danger to life. [ABSTRACT FROM AUTHOR]

The reproducibility of embryonic development is remarkable, although molecular processes are intrinsically stochastic at the single-cell level. How the multicellular system resists the inevitable noise to acquire developmental reproducibility constitutes a fundamental question in developmental biology. Toward this end, we focused on vertebrate somitogenesis as a representative system, because somites are repeatedly reproduced within a single embryo whereas such reproducibility is lost in segmentation clock gene-deficient embryos. However, the effect of noise on developmental reproducibility has not been fully investigated, because of the technical difficulty in manipulating the noise intensity in experiments. In this study, we developed a computational model of ERK-mediated somitogenesis, in which bistable ERK activity is regulated by an FGF gradient, cell-cell communication, and the segmentation clock, subject to the intrinsic noise. The model simulation generated our previous in vivo observation that the ERK activity was distributed in a step-like gradient in the presomitic mesoderm, and its boundary was posteriorly shifted by the clock in a stepwise manner, leading to regular somite formation. Here, we showed that this somite regularity was robustly maintained against the noise. Removing the clock from the model predicted that the stepwise shift of the ERK activity occurs at irregular timing with irregular distance owing to the noise, resulting in somite size variation. This model prediction was recently confirmed by live imaging of ERK activity in zebrafish embryos. Through theoretical analysis, we presented a mechanism by which the clock reduces the inherent somite irregularity observed in clock-deficient embryos. Therefore, this study indicates a novel role of the segmentation clock in noise-resistant developmental reproducibility. [ABSTRACT FROM AUTHOR]

The superorder Rhabdomoplea is established for copepods belonging to the order Thaumatopsylloida. Apomorphies for Rhabdomoplea are an adult prosome, including thoracic somites 1-4, complete at copepodid I, development of the posterior articulation of thoracic somite 7 of males delayed to copepodid IV and of abdominal somite 1 delayed to copepodid V, and absence of a posterior articulation of abdominal somites 2-3 forming with the anal somite a rod-like somite complex as part of the urosome. Rhabdomoplea appears to be the earliest branch of copepods because only thoracic somites 1-4 are broad throughout copepodid development, and thoracic somites 5 and 6 remain narrow. On podopleans and gymnopleans thoracic somite 5 is transformed from the anterior narrow somite to the posterior broad somite during the moult to copepodid II. On gymnopleans thoracic somite 6 also is transformed from the anterior narrow somite to the posterior broad somite but during the moult to copepodid III. Thus rhabdomopleans differ from podopleans in their body architecture as much as podopleans differ from gymnopleans. An alternative and traditional phylogeny that posits gymnopleans as the earliest branch of copepods requires reversal of these two transformations during copepod evolution; this hypothesis is not favoured here because the parsimonious hypothesis of direct, progressive transformations seems reasonable and plausible. [ABSTRACT FROM AUTHOR]

Abstract The enteric nervous system is mostly derived from vagal neural crest (NC) cells adjacent to somites (s)1–7. We used in ovo focal fluorescent vital dyes and focal electroporation of fluorophore-encoding plasmids in quail embryos to investigate NC cell migration to the foregut initially and later throughout the entire gut. NC cells of different somite-level origins were largely separate until reaching the foregut at about QE2.5, when all routes converged. By QE3.5, NC cells of different somite-levels became mixed, although s1-s2 NC cells were mainly confined to rostral foregut. Mid-vagal NC-derived cells (s3 and s4 level) arrived earliest at the foregut, and occurred in greatest number. By QE6.5 ENS was present from foregut to hindgut. Mid-vagal NC-derived cells occurred in greatest numbers from foregut to distal hindgut. NC-derived cells of s2, s5, and s6 levels were fewer and were widely distributed but were never observed in the distal hindgut. Rostro-vagal (s1) and caudo-vagal (s7) levels were few and restricted to the foregut. Single somite levels of quail neural tube/NC from s1 to s8 were combined with chick aneural ChE4.5 midgut and hindgut and the ensemble was grown on the chorio-allantoic membrane for 6 days. This tests ENS-forming competence in the absence of intra-segmental competition between NC cells, of differential influences of segmental paraxial tissues, and of positional advantage. All vagal NC-levels, but not s8 level, furnished enteric plexuses in the recipient gut, but the density of both ENS cells in total and neurons was highest from mid-vagal level donors, as was the length colonised. We conclude that the fate and competence for ENS formation of vagal NC sub-levels is not uniform over the vagal level but is biased to favour mid-vagal levels. Overviewing this and prior studies suggests the vagal region is, as in its traditional sense, a natural unit but with complex sub-divisions. Highlights • Neural crest cells of somite level s2 to s6 are fated to contribute to the ENS throughout the gut of avian embryos. • Neural crest cells of s3 and s4 levels have the most extensive and numerically largest contribution to the ENS. • Neural crest cells of somite levels s1 and s7 contribute ENS only to the foregut. • The capability of isolated somite levels of neural crest to form ENS in experiments exactly parallels the fate pattern. • The vagal neural crest level with somites s1 to s7 is a natural unit with complex axial pre-specification. [ABSTRACT FROM AUTHOR]

During analysis of sediment samples from South China Sea, a new species belonging to the genus Rhyncholagena Lang, 1944 was found and described here. Rhyncholagena paraspinifer sp. n. differs from its congeners by the following combined characteristics: body ornamented dorsally with at least one row of spinules on each somite except penultimate urosomite; A2 exopod two-segmented; P1 enp-2 with one inner seta; P3 exp-3 with two inner setae, P3 enp-2 with one inner seta; female P5 exopod with five setae; male P5 baseoendopod with two setae and exopod with four setae. This is the first report of the genus Rhyncholagena in the China seas. In addition, a key to all valid species of Rhyncholagena is given, along with tables of morphological characters of all valid species and their distributions. [ABSTRACT FROM AUTHOR]

HOX proteins act during development to regulate musculoskeletal morphology. HOXA5 patterns skeletal structures surrounding the cervical-thoracic transition including the vertebrae, ribs, sternum and forelimb girdle. However, the tissue types in which it acts to pattern the skeleton, and the ultimate fates of embryonic cells that activate Hoxa5 expression are unknown. A detailed characterization of HOXA5 expression by immunofluorescence was combined with Cre/LoxP genetic lineage tracing to map the fate of Hoxa5 expressing cells in axial musculoskeletal tissues and in their precursors, the somites and lateral plate mesoderm. HOXA5 protein expression is dynamic and spatially restricted in derivatives of both the lateral plate mesoderm and somites, including a subset of the lateral sclerotome, suggesting a local role in regulating early skeletal patterning. HOXA5 expression persists from somite stages through late development in differentiating skeletal and connective tissues, pointing to a continuous and direct role in skeletal patterning. In contrast, HOXA5 expression is excluded from the skeletal muscle and muscle satellite cell lineages. Furthermore, the descendants of Hoxa5-expressing cells, even after HOXA5 expression has extinguished, never contribute to these lineages. Together, these findings suggest cell autonomous roles for HOXA5 in skeletal development, as well as non-cell autonomous functions in muscle through expression in surrounding connective tissues. They also support the notion that different Hox genes display diverse tissue specificities and locations to achieve their patterning activity. [ABSTRACT FROM AUTHOR]

The mammalian body plan is shaped by rhythmic segmentation of mesoderm into somites, which are transient embryonic structures that form down each side of the neural tube. We have analyzed the genome-wide transcriptional and chromatin dynamics occurring within nascent somites, from early inception of somitogenesis to the latest stages of body plan establishment. We created matched gene expression and open chromatin maps for the three leading pairs of somites at six time points during mouse embryonic development. We show that the rate of somite differentiation accelerates as development progresses. We identified a conserved maturation program followed by all somites, but somites from more developed embryos concomitantly switch on differentiation programs from derivative cell lineages soon after segmentation. Integrated analysis of the somitic transcriptional and chromatin activities identified opposing regulatory modules controlling the onset of differentiation. Our results provide a powerful, high-resolution view of the molecular genetics underlying somitic development in mammals. [Display omitted] • Somite maturation follows a conserved molecular program across development • Somite differentiation accelerates with development • Somites at later development commit to derivative lineages soon after segmentation Ibarra-Soria and Thierion et al. characterize the molecular landscape of the three most recently segmented somites across six stages of development in mouse embryos. They find that while all somites follow a common maturation program, diversification into derivative lineages accelerates across development, occurring soon after segmentation in mature embryos. [ABSTRACT FROM AUTHOR]

In this study, the toxicity of sucrose to Oplegnathus punctatus embryos was evaluated. Embryos at the 4–6 somite, tail-bud, heart formation, and heart-beating stages were exposed to 0, 0.5, 1,1.5, 2, 2.5, or 3 M sucrose for 1 h. Survival rates of embryos at the tail-bud, heart formation, and heart-beating stages after rehydration for 1 h were not affected by treatment with 2 M sucrose (the maximum concentration). Embryos at the tail-bud, heart formation, and heart-beating stages were exposed to 2 M sucrose for 0, 30, 60, 90, 120, 150, or 180 min. Long-term developmental indicators, including rates of survival, hatching, swimming, and malformation, were evaluated for 4 days after rehydration. Based on the survival rates 10 min after rehydration, the longest tolerance time for embryos at the three stages was 120 min. Based on long-term developmental indicators, the longest tolerance times were 60 min at the tail-bud, 60 min at the heart formation stage and 30 min at the heart beating stage. The malformation rates increased as the treatment time increased. The malformation rates were 100% when embryos were exposed to sucrose for ≥120 min. Malformation was divided into larval and embryonic abnormality. As the exposure time increased for tail-bud stage embryos, the rate of larval malformation increased. Treatment at heart formation and heart-beating stages resulted in higher rates of failure to hatch at exposure time. Based on these results, toxicity tests of non-permeable cryoprotectant in embryos requires the observation of development for at least 2 days after rehydration. Based on long-term observation, it was concluded that dehydration before freezing was not the direct cause of larvae deformity that hatched from frozen-thawing embryo. These results provide a reference for the singly use of representative non-permeable cryoprotectant sucrose. • Toxicity test results for representative non-permeable cryoprotectant sucrose of Oplegnathus punctatus embryos was studied for the first time. • Observation for 2 days after rehydration is recommended for toxicity tests of non-permeable cryoprotectant. • Dehydration of tail-bud and heart-beating stage embryos using 2 M sucrose for 30 min caused deformity rate of hatched larvae lower than 0.8%. [ABSTRACT FROM AUTHOR]

Zebrafish are a valuable vertebrate model in which to study development and characterize genes involved in cystic kidney disease. Zebrafish embryos and larvae are transparent, allowing non-invasive imaging during their rapid development, which takes place over the first 72 hours post fertilisation. Gene-specific knockdown of nephronophthisis-associated genes leads to ciliary phenotypes which can be assessed in various developmental structures. Here we describe in detail the methods used for imaging cilia within Kupffer’s vesicle to assess nephronophthisis and related ciliopathy phenotypes.

Heterogeneity of stem cells or their niches is likely to influence tissue regeneration. Here we reveal stem/precursor cell diversity during wound repair in larval zebrafish somitic body muscle using time-lapse 3D confocal microscopy on reporter lines. Skeletal muscle with incision wounds rapidly regenerates both slow and fast muscle fibre types. A swift immune response is followed by an increase in cells at the wound site, many of which express the muscle stem cell marker Pax7. Pax7+ cells proliferate and then undergo terminal differentiation involving Myogenin accumulation and subsequent loss of Pax7 followed by elongation and fusion to repair fast muscle fibres. Analysis of pax7a and pax7b transgenic reporter fish reveals that cells expressing each of the duplicated pax7 genes are distinctly localised in uninjured larvae. Cells marked by pax7a only or by both pax7a and pax7b enter the wound rapidly and contribute to muscle wound repair, but each behaves differently. Low numbers of pax7a-only cells form nascent fibres. Time-lapse microscopy revealed that the more numerous pax7b-marked cells frequently fuse to pre-existing fibres, contributing more strongly than pax7a-only cells to repair of damaged fibres. pax7b-marked cells are more often present in rows of aligned cells that are observed to fuse into a single fibre, but more rarely contribute to nascent regenerated fibres. Ablation of a substantial portion of nitroreductase-expressing pax7b cells with metronidazole prior to wounding triggered rapid pax7a-only cell accumulation, but this neither inhibited nor augmented pax7a-only cell-derived myogenesis and thus altered the cellular repair dynamics during wound healing. Moreover, pax7a-only cells did not regenerate pax7b cells, suggesting a lineage distinction. We propose a modified founder cell and fusion-competent cell model in which pax7a-only cells initiate fibre formation and pax7b cells contribute to fibre growth. This newly discovered cellular complexity in muscle wound repair raises the possibility that distinct populations of myogenic cells contribute differentially to repair in other vertebrates.

The patterning of the spine of a zebrafish is controlled by the notochord, a rod-like structure that supports and instructs the developing embryo.

Segmentation of the axial skeleton in amniotes depends on the segmentation clock, which patterns the paraxial mesoderm and the sclerotome. While the segmentation clock clearly operates in teleosts, the role of the sclerotome in establishing the axial skeleton is unclear. We severely disrupt zebrafish paraxial segmentation, yet observe a largely normal segmentation process of the chordacentra. We demonstrate that axial entpd5+ notochord sheath cells are responsible for chordacentrum mineralization, and serve as a marker for axial segmentation. While autonomous within the notochord sheath, entpd5 expression and centrum formation show some plasticity and can respond to myotome pattern. These observations reveal for the first time the dynamics of notochord segmentation in a teleost, and are consistent with an autonomous patterning mechanism that is influenced, but not determined by adjacent paraxial mesoderm. This behavior is not consistent with a clock-type mechanism in the notochord.

Abstract: Some vertebral anomalies in cultured fish arise from unusual environmental conditions during embryogenesis. We investigated the individual and combined teratogenic effects of short‐term hypercapnia and hypoxia on embryos of red sea bream (RSB), Pagrus major. Ten‐somite stage embryos were exposed to hypercapnia (60 and 120 mg/L dissolved carbon dioxide: DCD) and hypoxia (0%, 10%, 25%, 50%, and 75% dissolved oxygen: DO) independently and concomitantly for seven different periods (0, 30, 60, 90, 120, 150 and 180 min) to examine somitic disturbances at hatching, which are prodromal symptoms of centrum defects. Another experiment examined vertebral anomalies in juveniles raised from eggs exposed to hypercapnia (120 mg/L DCD) and hypoxia (10% DO) independently and concomitantly. Short‐time exposures (30–180 min) to hypercapnia (60 and 120 mg/L) and hypoxia (10% DO or less) independently and additively caused larval somitic disturbances and juvenile centrum defects. The results indicate that short‐term hypercapnia and hypoxia generally have the same teratogenic effect on embryos, although there were some differences in the locations of the somitic disturbances and centrum defects, with additive teratogenicity when the conditions were combined. These results suggest the necessity of maintaining appropriate DCD and DO concentrations during egg incubation and transport and during reproduction in recirculating aquaculture. [ABSTRACT FROM AUTHOR]

Mesoderm is derived from the primitive streak. The rostral region of the primitive streak forms the somitic mesoderm. We have previously shown the developmental origin of each level of the somitic mesoderm using DiI fluorescence labeling of the primitive streak. We found that the more caudal segments were derived from the primitive streak during the later developmental stages. DiI labeled several pairs of somites and showed the distinct rostral boundary; however, the fluorescence gradually disappeared in the caudal region. This finding can be explained in two ways: the primitive streak at a specific developmental stage is primordial of only a certain number of pairs of somites, or the DiI fluorescent dye was gradually diluted within the primitive streak by cell division. Here, we traced the development of the primitive streak cells using enhanced green fluorescent protein (EGFP) transfection. We confirmed that, the later the EGFP transfection stage, the more caudal the somites labeled. Different from DiI labeling, EGFP transfection performed at any developmental stage labeled the entire somitic mesoderm from the anterior boundary to the tail bud in 4.5-day-old embryos. Furthermore, the secondary neural tube was also labeled, suggesting that not only the somite precursor cells but also the axial stem cells were labeled. [ABSTRACT FROM AUTHOR]

The diaphragm is a mammalian skeletal muscle essential for respiration and for separating the thoracic and abdominal cavities. Development of the diaphragm requires the coordinated development of muscle, muscle connective tissue, tendon, nerves, and vasculature that derive from different embryonic sources. However, defects in diaphragm development are common and the cause of an often deadly birth defect, Congenital Diaphragmatic Hernia (CDH). Here we comprehensively describe the normal developmental origin and complex spatial-temporal relationship between the different developing tissues to form a functional diaphragm using a developmental series of mouse embryos genetically and immunofluorescently labeled and analyzed in whole mount. We find that the earliest developmental events are the emigration of muscle progenitors from cervical somites followed by the projection of phrenic nerve axons from the cervical neural tube. Muscle progenitors and phrenic nerve target the pleuroperitoneal folds (PPFs), transient pyramidal-shaped structures that form between the thoracic and abdominal cavities. Subsequently, the PPFs expand across the surface of the liver to give rise to the muscle connective tissue and central tendon, and the leading edge of their expansion precedes muscle morphogenesis, formation of the vascular network, and outgrowth and branching of the phrenic nerve. Thus development and morphogenesis of the PPFs is critical for diaphragm formation. In addition, our data indicate that the earliest events in diaphragm development are critical for the etiology of CDH and instrumental to the evolution of the diaphragm. CDH initiates prior to E12.5 in mouse and suggests that defects in the early PPF formation or their ability to recruit muscle are an important source of CDH. Also, the recruitment of muscle progenitors from cervical somites to the nascent PPFs is uniquely mammalian and a key developmental innovation essential for the evolution of the muscularized diaphragm. [ABSTRACT FROM AUTHOR]

Cadmium is one of the most toxic metals that has high environmental stability and can hardly be converted into substances of low toxicity. In the present study, the effects of cadmium on the development of somites of chick embryos under in vitro conditions were investigated. First, fertilized eggs were incubated until they reached the stage of 15-20 somites. The embryos were then separated from the yolk by using the window technique. The somites were separated by insulin needles under a stereomicroscope. The separated somites were then transferred to Dulbecco's modified Eagle's medium (DMEM) stock culture and treated with different concentrations of cadmium nitrate (0, 500, 1000, 2000, and 4000 ng/mL). After 72 h, the diameter of the nucleus, cell morphology, and extracellular matrix compounds were examined. Our results indicated that cadmium disrupted the developmental process of the somites. The microscopic studies of the somites, based on a histological technique, indicated that cadmium induced apoptosis in somite cells and also decreased the chondrogenesis and collagen synthesis. Therefore, somites are considered suitable temporary embryonic organs and interesting models for basic developmental study under in vitro conditions. Moreover, it might be possible to use somites in vitro for studying the effects of trace elements on living organisms. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs) are highly conserved small non-coding RNA molecules that post-transcriptionally regulate gene expression in multicellular organisms. Within the set of muscle-specific miRNAs, miR-206 expression is largely restricted to skeletal muscle and is found exclusively within the bony fish lineage. Although many studies have implicated miR-206 in muscle maintenance and disease, its role in skeletal muscle development remains largely unknown. Here, we examine the role of miR-206 during Xenopus laevis somitogenesis. In Xenopus laevis, miR-206 expression coincides with the onset of somitogenesis. We show that both knockdown and over-expression of miR-206 result in abnormal somite formation affecting muscle cell rotation, attachment, and elongation. In particular, our data suggests that miR-206 regulates changes in cell adhesion that affect the ability of newly formed somites to adhere to the notochord as well as to the intersomitic boundaries. Additionally, we show that β-dystroglycan and F-actin expression levels are significantly reduced, suggesting that knockdown of miR-206 levels affects cellular mechanics necessary for cell shape changes and attachments that are required for proper muscle formation. [ABSTRACT FROM AUTHOR]

Abstract: Although the human tail is completely absent at birth, the embryonic tail is formed just as in other tailed amniotes. Since all morphological variations are created from variations in developmental processes, elucidation of the tail reduction process during embryonic development may be necessary to clarify the human evolutionary process. The tail has also been of great interest to the medical community. The congenital anomaly referred to as ‘human tail’, i.e. the occurrence of a tail‐like structure, has been reported and was thought to represent a vestige of the embryonic tail; however, this hypothesis has not been verified. Accordingly, in this study, we aimed to establish a new method to visualize all somites in an embryo. We used sagittal‐sectioned embryos from Carnegie Stage (CS) 13 to CS23. All samples were obtained from the Congenital Anomaly Research Center, Kyoto University, Japan. Combining photomicroscopy and three‐dimensional reconstruction, we clearly visualized and labeled all somites. We found that the number of somites peaked at CS16 and dramatically decreased by approximately five somites. Tail reduction with a decrease in somites has also been observed in other short‐tailed amniotes; thus, this result suggested the possibility that there is a common mechanism for morphogenesis of short tails in amniote species. Additionally, our findings provided important insights into the cause of the congenital anomaly known as ‘human tail’. [ABSTRACT FROM AUTHOR]

One of the definitive characteristics of chordates (cephalochordates, vertebrates) is the somites, which are a series of paraxial mesodermal blocks exhibiting segmentation. The presence of somites in the basal chordate amphioxus and in vertebrates, but not in tunicates (the sister group of vertebrates), suggests that the tunicates lost the somites secondarily. Somites are patterned from anterior to posterior during embryogenesis. How such a segmental pattern evolved from deuterostome ancestors is mysterious. The classic enterocoel theory claims that chordate mesoderm evolved from the ancestral deuterostome mesoderm that organizes the trimeric body parts seen in extant hemichordates. Recent progress in molecular embryology has been tremendous, which has enabled us to test this classic theory. In this review, the history of the study on the evolution of the chordate mesoderm is summarized. This is followed by a review of the current understanding of genetic mapping on anterior/posterior (A/P) mesodermal patterning between chordates (cephalochordates, vertebrates) and a direct developing hemichordate (Saccoglossus kowalevskii). Finally, a possible scenario about the evolution of the chordate mesoderm from deuterostome ancestors is discussed. [ABSTRACT FROM AUTHOR]

A prominent anatomical feature of the peripheral nervous system is the segmentation of mixed (motor, sensory and autonomic) spinal nerves alongside the spinal cord. During early development their axon growth cones avoid the developing vertebral elements by traversing the anterior/cranial half of each somite-derived sclerotome, so ensuring the separation of spinal nerves from vertebral bones as axons extend towards their peripheral targets. A glycoprotein expressed on the surface of posterior half-sclerotome cells confines growth cones to the anterior half-sclerotomes by contact repulsion. A closely similar glycoprotein is expressed in avian and mammalian grey matter, where we hypothesize it may have evolved to regulate neural plasticity in birds and mammals. [ABSTRACT FROM AUTHOR]

Objectives: Three issues are considered on variation in number of presacral vertebrae (PSV) in humans: (1) sexual difference in number of PSV, (2) inactivation of Hoxd-11 gene as etiology for a supernumerary lumbar vertebra, and (3) anatomical correlates of a supernumerary lumbar vertebra, including lumbar-sacral nearthrosis, and pelvic size. Materials and methods: Sample was 407 skeletonized females and 1,318 males from United States; ages at death were 20 to 49 years. Two subsamples of males were used: (1) 98 with modal numbers of cervical, thoracic, lumbar, and sacral vertebrae (PSV=24) and (2) 45 with a supernumerary lumbar vertebra but modal numbers for other vertebral segments (PSV=25). Measurements were taken of ulna, second metacarpal, vertebrae, femur, and pelvis; presence of lumbar-sacral nearthrosis was observed. Results: Although 90% of females and males have 24 PSV, females have higher frequency of 23 PSV and males have higher frequency of 25 PSV. Compared to males with 24 PSV, males with 25 PSV and supernumerary lumbar vertebra show (1) no difference in anatomies associated with inactivation of Hoxd-11, and (2) higher frequency of lumbar-sacral nearthrosis and smaller pelvic inlet circumference. Discussion: Sexual difference in number of PSV may be due to tempo of somite formation and Hox gene activation. Hypothesis is not supported that a supernumerary lumbar vertebra is due to inactivation of Hoxd-11. The presence of a supernumerary lumbar vertebra is associated with small pelvic inlet circumference, which can be obstetrically disadvantageous. [ABSTRACT FROM AUTHOR]

The ADAMTS5 metzincin, a secreted zinc-dependent metalloproteinase, modulates the extracellular matrix (ECM) during limb morphogenesis and other developmental processes. Here, the role of ADAMTS5 was investigated by knockdown of zebrafish adamts5 during embryogenesis. This revealed impaired Sonic Hedgehog (Shh) signaling during somite patterning and early myogenesis. Notably, synergistic regulation of myod expression by ADAMTS5 and Shh during somite differentiation was observed. These roles were not dependent upon the catalytic activity of ADAMTS5. These data identify a non-enzymatic function for ADAMTS5 in regulating an important cell signaling pathway that impacts on muscle development, with implications for musculoskeletal diseases in which ADAMTS5 and Shh have been associated. [ABSTRACT FROM AUTHOR]

Introduction. Klippel–Feil syndrome (KFS) is a congenital anomaly resulting from fusion of cervical vertebral bodies secondary to the dysregulation of signaling pathways during somite development. It is commonly associated with scoliosis and Sprengel deformity. We present a case of KFS with commonly associated abnormalities as well as deformities that have not yet been reported in the literature. Case Presentation. A 3-year-old girl presented for further evaluation of a left upper extremity deformity following a negative genetic workup. Upon physical exam and radiographic imaging, the patient was diagnosed with KFS and associated abnormalities including cervical scoliosis, Sprengel deformity, and congenital deformity of the left upper extremity. Deformities of the left upper extremity include radioulnar synostosis, a four-rayed hand, and absent thenar musculature. The Sprengel deformity was corrected surgically with a Woodward procedure. Discussion. Congenital musculoskeletal deformities can be differentiated based upon spinal and limb embryology. The presence of extraspinal abnormalities not originating from somite differentiation may suggest a severe form of KFS. Important considerations in the workup of the KFS patient include looking for deformities of the shoulder girdle and upper extremities to identify abnormalities for intervention at a young age. [ABSTRACT FROM AUTHOR]

The chick embryo has provided a prominent model system for the study of segmental patterning in the nervous system. During early development, motor and sensory axon growth cones traverse the anterior/rostral half of each somite, so avoiding the developing vertebral components and ensuring separation of spinal nerves from vertebral bones. A glycoprotein expressed on the surface of posterior half-somite cells confines growth cones to the anterior half-somites by a contact repulsive mechanism. Hindbrain segmentation is also a conspicuous feature of chick brain development. We review how its contemporary analysis was initiated in the chick embryo, and the advantages the chick system continues to provide in its detailed elucidation at both molecular and neural circuit levels. [ABSTRACT FROM AUTHOR]

Striated muscle is the most abundant tissue in the body of vertebrates and it forms, together with the skeleton, the locomotory system required both for movement and the creation of the specific body shape of a species. Research on the embryonic development of muscles has a long tradition both in classical embryology and in molecular developmental biology. While the gene networks regulating muscle development have been discovered mostly in the mouse through genetics, our knowledge on cell lineages, muscle morphogenesis and tissue interactions regulating their formation is to a large extent based on the use of the avian model. This review highlights present knowledge of the development of skeletal muscle in vertebrate embryos. Special focus will be placed on the contributions from chicken and quail embryo model systems. [ABSTRACT FROM AUTHOR]