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The metameric pattern of somites is created based on oscillatory expression of clock genes in presomitic mesoderm. However, the mechanism for converting the dynamic oscillation to a static pattern of somites is still unclear. Here, we provide evidence that Ripply/Tbx6 machinery is a key regulator of this conversion. Ripply1/Ripply2-mediated removal of Tbx6 protein defines somite boundary and also leads to cessation of clock gene expression in zebrafish embryos. On the other hand, activation of ripply1/ripply2 mRNA and protein expression is periodically regulated by clock oscillation in conjunction with an Erk signaling gradient. Whereas Ripply protein decreases rapidly in embryos, Ripply-triggered Tbx6 suppression persists long enough to complete somite boundary formation. Mathematical modeling shows that a molecular network based on results of this study can reproduce dynamic-to-static conversion in somitogenesis. Furthermore, simulations with this model suggest that sustained suppression of Tbx6 caused by Ripply is crucial in this conversion. During somitogenesis, the dynamic oscillation of the molecular clock is converted into static spatial patterns. Here, the authors show that persistent suppression of Tbx6 expression triggered by periodical Ripply1/2 gene expression is a key to this conversion. [ABSTRACT FROM AUTHOR]

During embryonic development, epithelial cell blocks called somites are periodically formed according to the segmentation clock, becoming the foundation for the segmental pattern of the vertebral column. The process of somitogenesis has recently been recapitulated with murine and human pluripotent stem cells. However, an in vitro model for human somitogenesis coupled with the segmentation clock and epithelialization is still missing. Here, we report the generation of human somitoids, organoids that periodically form pairs of epithelial somite-like structures. Somitoids display clear oscillations of the segmentation clock that coincide with the segmentation of the presomitic mesoderm. The resulting somites show anterior-posterior and apical-basal polarities. Matrigel is essential for epithelialization but dispensable for the differentiation into somite cells. The size of somites is rather constant, irrespective of the initial cell number. The amount of WNT signaling instructs the proportion of mesodermal lineages in somitoids. Somitoids provide a novel platform to study human somitogenesis. Somitogenesis has been well characterized in model organisms, resulting in detailed description of the somite segmentation clock. Here they generate somitogenic organoids from human pluripotent stem cells that recapitulate somitogenesis, periodic segmentation, and proper polarity. [ABSTRACT FROM AUTHOR]

Somitogenesis, the segmentation of the antero-posterior axis in vertebrates, is thought to result from the interactions between a genetic oscillator and a posterior-moving determination wavefront. The segment (somite) size is set by the product of the oscillator period and the velocity of the determination wavefront. Surprisingly, while the segmentation period can vary by a factor three between 20 °C and 32 °C, the somite size is constant. How this temperature independence is achieved is a mystery that we address in this study. Using RT-qPCR we show that the endogenous fgf8 mRNA concentration decreases during somitogenesis and correlates with the exponent of the shrinking pre-somitic mesoderm (PSM) size. As the temperature decreases, the dynamics of fgf8 and many other gene transcripts, as well as the segmentation frequency and the PSM shortening and tail growth rates slows down as T–Tc (with Tc = 14.4 °C). This behavior characteristic of a system near a critical point may account for the temperature independence of somitogenesis in zebrafish. In Zebrafish, the dynamics of fgf8 and other gene transcripts as well as segmentation frequency, shortening of pre-somitic mesoderm and tail growth rate slows down with lower temperature. This may explain the temperature independence of somitogenesis. [ABSTRACT FROM AUTHOR]

Complex systems, ranging from developing embryos to systems of locally communicating agents, display an apparent capability of "programmable" pattern formation: They reproducibly form target patterns, but those targets can be readily changed. A distinguishing feature of such systems is that their subunits are capable of information processing. Here, we explore schemes for programmable pattern formation within a theoretical framework, in which subunits process local signals to update their discrete state following logical rules. We study systems with different update rules, topologies, and control schemes, assessing their capability of programmable pattern formation and their susceptibility to errors. Only a fraction permits local organizers to dictate any target pattern, by transcribing temporal patterns into spatial patterns, reminiscent of the principle underlying vertebrate somitogenesis. An alternative scheme employing variable rules cannot reach all patterns but is insensitive to the timing of organizer inputs. Our results establish a basis for designing synthetic systems and models of programmable pattern formation closer to real systems. Pattern formation in complex systems has been studied from the angle of information processing, by trying to understand what is the input information that determines a certain final pattern and to what extent is the final pattern programmable by changing some input information. Here, the authors present a minimal model within which they can formulate and study the concept of programmable pattern formation, that is, whether an organizer cell can steer the system towards any target pattern starting from an arbitrary initial pattern. [ABSTRACT FROM AUTHOR]

During embryogenesis, the processes that control how cells differentiate and interact to form particular tissues and organs with precise timing and shape are of fundamental importance. One prominent example of such processes is vertebrate somitogenesis, which is governed by a molecular oscillator called the segmentation clock. The segmentation clock system is initiated in the presomitic mesoderm in which a set of genes and signaling pathways exhibit coordinated spatiotemporal dynamics to establish regularly spaced boundaries along the body axis; these boundaries provide a blueprint for the development of segment-like structures such as spines and skeletal muscles. The highly complex and dynamic nature of this in vivo event and the design principles and their regulation in both normal and abnormal embryogenesis are not fully understood. Recently, live-imaging has been used to quantitatively analyze the dynamics of selected components of the circuit, particularly in combination with well-designed experiments to perturb the system. Here, we review recent progress from studies using live imaging and manipulation, including attempts to recapitulate the segmentation clock in vitro. In combination with mathematical modeling, these techniques have become essential for disclosing novel aspects of the clock. [ABSTRACT FROM AUTHOR]

An elongated and segmented body plan is a common morphological characteristic of all arthropods and is probably responsible for their high adaptation ability to diverse environments. Most arthropods form their bodies by progressively adding segments, resembling vertebrate somitogenesis. This sequential segmentation relies on a molecular clock that operates in the posterior region of the elongating embryo that combines dynamically with cellular behaviors and tissue rearrangements, allowing the extension of the developing body along its main embryonic axis. Even though the molecular mechanisms involved in elongation and segment formation have been found to be conserved in a considerable degree, cellular processes such as cell division are quite variable between different arthropods. In this study, we show that cell proliferation in the beetle Tribolium castaneum has a nonuniform spatiotemporal patterning during axial elongation. We found that dividing cells are preferentially oriented along the anterior–posterior axis, more abundant and posteriorly localized during thoracic segments formation and that this cell proliferation peak was triggered at the onset of axis elongation. This raise in cell divisions, in turn, was correlated with an increase in the elongation rate, but not with changes in cell density. When DNA synthesis was inhibited over this period, both the area and length of thoracic segments were significantly reduced but not of the first abdominal segment. We discuss the variable participation that different cell division patterns and cell movements may have on arthropod posterior growth and their evolutionary contribution. [ABSTRACT FROM AUTHOR]

A quantitative description of the molecular networks that sustain morphogenesis is one of the main challenges of developmental biology. In particular, a molecular understanding of the segmentation of the antero-posterior axis in vertebrates has yet to be achieved. This process known as somitogenesis is believed to result from the interactions between a well-studied genetic oscillator and a less established posterior-moving determination wavefront. Here we describe a molecular model for somitogenesis that couples a moving morphogen wavefront with the somitogenetic oscillator. The wavefront is due to a switch between stable states that results from reciprocal negative feedbacks of Retinoic Acid (RA) on the activation of a kinase ErK and of ErK on RA synthesis. We suggest a molecular mechanism by which that switch can be triggered by the somitogenetic clock. The model quantitatively accounts for the shortening of the pre-somitic mesoderm (PSM) in zebrafish in response to the decrease during somitogenesis in the concentration of a morphogen (Fgf8). The generality and robustness of the model allows for its validation (or invalidation) in other model organisms. [ABSTRACT FROM AUTHOR]

In this paper, we investigated the modified two dimensional model which can explain somite patterning in embryos. It is suitable for exploring a design space of somitogenesis and can explain aspects of somitogenesis that previous models cannot. Here, we mainly studied the non-diffusing case. We have used the Hopf bifurcation theorem, the Center manifold theorem and Runge-Kutta method in our investigation. First, we investigate its dynamical behaviors and put forward a sufficient condition for the oscillation of the small network. Then, we give the mathematical simulation based on the Runge-Kutta method. In the process of solving ordinary differential equations, the four order Runge-Kutta method has the advantages of high accuracy, convergence and stability (under certain conditions), which can change the step size and do not need to calculate higher order derivatives. Therefore, it has become the most commonly used numerical solution. At the same time, we get the sufficient condition in which the bistable state of the system exists and give the numerical simulation. Because somitogenesis occupies an important position in the process of biological development, and as a pattern process can be used to study pattern formation and many aspects of embryogenesis. So our study have a great help for embryonic development, gene expression, cell differentiation. In addition, it is beneficial to study the clone animal variation problem of spinal bone number and is of great help to the treatment and prevention of defects of human spine disease. [ABSTRACT FROM AUTHOR]

Background: Cardiac anomalies are prevalent in patients with bony spinal anomalies. Prior studies evaluating incidences of bony congenital anomalies of the spine are limited. The Kids’ Inpatient Database (KID) yields national discharge estimates of rare pediatric conditions like congenital disorders. This study utilized cluster analysis to study patterns of concurrent vertebral anomalies, anal atresia, cardiac malformations, trachea-esophageal fistula, renal dysplasia, and limb anomalies (VACTERL anomalies) co-occurring in patients with spinal congenital anomalies.Methods: Retrospective review of KID 2003-2012. KID-supplied hospital- and year-adjusted weights allowed for incidence assessment of bony spinal anomalies and cardiac, gastrointestinal, urinary anomalies of VACTERL. K-means clustering assessed relationships between most frequent anomalies within bony spinal anomaly discharges; k set to n − 1(n = first incidence of significant drop/little gain in sum of square errors within clusters).Results: There were 12,039,432 KID patients 0-20 years. Incidence per 100,000 discharges: 2.5 congenital fusion of spine, 10.4 hemivertebra, 7.0 missing vertebra. The most common anomalies co-occurring with bony vertebral malformations were atrial septal defect (ASD 12.3%), large intestinal atresia (LIA 11.8%), and patent ductus arteriosus (PDA 10.4%). Top congenital cardiac anomalies in vertebral anomaly patients were ASD, PDA, and ventricular septal defect (VSD); all three anomalies co-occur at 6.6% rate in this vertebral anomaly population. Cluster analysis revealed that of bony anomaly discharges, 55.9% of those with PDA had ASD, 34.2% with VSD had PDA, 22.9% with LIA had ASD, 37.2% with ureter obstruction had LIA, and 35.5% with renal dysplasia had LIA.Conclusions: In vertebral anomaly patients, the most common co-occurring congenital anomalies were cardiac, renal, and gastrointestinal. Top congenital cardiac anomalies in vertebral anomaly patients were ASD, PDA, and VSD. VACTERL patients with vertebral anomalies commonly presented alongside cardiac and renal anomalies. [ABSTRACT FROM AUTHOR]

During early embryonic development, the vertebrate main body axis is segmented from head-to-tail into somites. Somites emerge sequentially from the presomitic mesoderm (PSM) as a consequence of oscillatory waves of genetic activity, called somitogenesis waves. Here, we discuss the implications of the dynamic patterns of early X-Delta-2 expression in the prospective somites (somitomeres) of Xenopus laevis. We report that right somitomeres normally emerge before left to form chiral structures (i.e. structures having clockwise or counter-clockwise handedness). From our observations, we infer that somitogenesis waves are normally counter-clockwise spirals, a novel dynamic mechanism for the control of handedness development in Xenopus. We propose that the same mechanism could control handedness development in all vertebrate embryos, providing a dynamical basis for the current asymmetric molecular transport model for generating left-right asymmetry. [ABSTRACT FROM AUTHOR]

Somitogenesis is the process for the development of somites in vertebrate embryos. This process is timely regulated by synchronous oscillatory expression of the segmentation clock genes. Mathematical models expressed by delay equations or ODEs have been proposed to depict the kinetics of these genes in interacting cells. Through mathematical analysis, we investigate the parameter regimes for synchronous oscillations and oscillation-arrested in an ODE model and a model with transcriptional and translational delays, both with Michaelis-Menten type degradations. Comparisons between these regimes for the two models are made. The delay model has larger capacity to accommodate synchronous oscillations. Based on the analysis and numerical computations extended from the analysis, we explore how the periods and amplitudes of the oscillations vary with the degradation rates, synthesis rates, and coupling strength. For typical parameter values, the period and amplitude increase as some synthesis rate or the coupling strength increases in the ODE model. Such variational properties of oscillations depend also on the magnitudes of time delays in delay model. We also illustrate the difference between the dynamics in systems modeled with linear degradation and the ones in systems with Michaelis-Menten type reactions for the degradation. The chief concerns are the connections between the dynamics in these models and the mechanism for the segmentation clocks, and the pertinence of mathematical modeling on somitogenesis in zebrafish. [ABSTRACT FROM AUTHOR]

In the grass snake ( Natrix natrix), the newly developed somites form vesicles that are located on both sides of the neural tube. The walls of the vesicles are composed of tightly connected epithelial cells surrounding the cavity (the somitocoel). Also, in the newly formed somites, the Pax3 protein can be observed in the somite wall cells. Subsequently, the somite splits into three compartments: the sclerotome, dermomyotome (with the dorsomedial [DM] and the ventrolateral [VL] lips) and the myotome. At this stage, the Pax3 protein is detected in both the DM and VL lips of the dermomyotome and in the mononucleated cells of the myotome, whereas the Pax7 protein is observed in the medial part of the dermomyotome and in some of the mononucleated cells of the myotome. The mononucleated cells then become elongated and form myotubes. As myogenesis proceeds, the myotome is filled with multinucleated myotubes accompanied by mononucleated, Pax7-positive cells (satellite cells) that are involved in muscle growth. The Pax3-positive progenitor muscle cells are no longer observed. Moreover, we have observed unique features in the differentiation of the muscles in these snakes. Specifically, our studies have revealed the presence of two classes of muscles in the myotomes. The first class is characterised by fast muscle fibres, with myofibrils equally distributed throughout the sarcoplasm. In the second class, composed of slow muscle fibres, the sarcoplasm is filled with lipid droplets. We assume that their storage could play a crucial role during hibernation in the adult snakes. We suggest that the model of myotomal myogenesis in reptiles, birds and mammals shows the same morphological and molecular character. We therefore believe that the grass snake, in spite of the unique features of its myogenesis, fits into the amniotes-specific model of trunk muscle development. [ABSTRACT FROM AUTHOR]

Abstract Segmentation is a common feature of disparate clades of metazoans, and its evolution is a central problem of evolutionary developmental biology. We evolved in silico regulatory networks by a mutation/selection process that just rewards the number of segment boundaries. For segmentation controlled by a static gradient, as in long‐germ band insects, a cascade of adjacent repressors reminiscent of gap genes evolves. For sequential segmentation controlled by a moving gradient, similar to vertebrate somitogenesis, we invariably observe a very constrained evolutionary path or funnel. The evolved state is a cell autonomous ‘clock and wavefront’ model, with the new attribute of a separate bistable system driven by an autonomous clock. Early stages in the evolution of both modes of segmentation are functionally similar, and simulations suggest a possible path for their interconversion. Our computation illustrates how complex traits can evolve by the incremental addition of new functions on top of pre‐existing traits.

Dmrt genes encode a large family of transcription factors characterized by the presence of a DM domain, an unusual zinc finger DNA binding domain. While Dmrt genes are well known for their important role in sexual development in arthropodes, nematodes and vertebrates, several new findings indicate emerging functions of this gene family in other developmental processes. Here, we provide an overview of the evolution, structure and mechanisms of action of Dmrt genes. We summarize recent findings on their function in sexual regulation and discuss more extensively the role played by these proteins in somitogenesis and neural development. [ABSTRACT FROM AUTHOR]

Introduction: Administration of cadmium (Cd) after 60 h (H) incubation induces ventral body wall defect (VBWD) similar to the omphalocele phenotype in the chick embryo. In this model, the earliest histological changes have been observed in somites commencing at 4-h post-treatment (4H). The molecular mechanism by which Cd acts in this critical period of embryogenesis still remains unclear. Sonic hedgehog (SHH) signalling plays an important role in vertebrate development, including somitogenesis and thus ventral body wall formation. Patched (PTCH), a cell membrane receptor for SHH, is expressed in somites and Patched knockout mice display somite dysfunction. Another transmembrane receptor, Smoothened (SMO), is also expressed in somites and transduces the SHH signal regulated by PTCH. We designed this study to test the hypothesis that SHH signalling is downregulated during the critical period of early embryogenesis in the Cd-induced omphalocele chick model. Methods: After 60 h of incubation, chicks were exposed to either chick saline or 50 μL of 50 μM cadmium acetate and divided into two groups: control and Cd ( n = 24 for each group). Chicks were harvested 1, 4, and 8 h post-treatment. Real-time RT-PCR was performed to evaluate the relative mRNA expression level of SHH, PTCH and SMO. Immunofluorescence confocal microscopy was then performed to evaluate protein expression/distribution of SHH, PTCH and SMO. Results: The relative mRNA expression levels of SHH, PTCH and SMO were significantly downregulated in the Cd group compared to controls at 4H post treatment, whereas, there were no significant differences at the other time points. Immunohistochemistry revealed that the intensity of SHH, PTCH and SMO was markedly diminished at 4 h in Cd-treated embryos compared to controls. Conclusion: Disturbance of the SHH signalling pathway as evidenced by SHH, PTCH and SMO downregulation during the narrow window of early embryogenesis may result in somite maldevelopment, contributing to the omphalocele phenotype in the Cd chick model. [ABSTRACT FROM AUTHOR]

Aim: In chick embryos, administration of cadmium (Cd) induces ventral body wall defects (VBWD) similar to human omphalocele. It has been shown that failure of proper VBW formation may be due to disruption of somite development during early embryogenesis. In the VBWD chick model, Cd causes abnormal cell death in the somitic region resulting in improperly developed somites and tortuosity of the neural tube. However, the exact molecular mechanisms leading to VBWD still remain unclear. Wnt signaling is crucial during embryogenesis and plays a key role in normal somite formation. The Rho-associated coiled-coil containing protein kinase (ROCK) is involved in the non-canonical Wnt pathway which controls actin cytoskeleton assembly and cell contractility, and contributes to several developmental processes including somitogenesis. ROCK gene expression levels have recently been reported to be significantly decreased in the Cd-induced VBWD chick model. We designed this study to investigate the hypothesis that administration of ROCK inhibitor (Y-27632) in the absence of Cd disrupts somitogenesis and could contribute to the development of VBWD during early embryogenesis. Methods: After 60 h of incubation chick embryos were transferred from eggs to culture dishes containing 20 μM of Y-27632 for experimental group (Y-27, n = 22) or chick saline for controls ( n = 14). Following 24 h in the incubator they were assessed for stage development and gross abnormalities in morphology using the dissecting microscope. Western blot was performed to confirm Y-27632 inhibition of ROCK downstream signaling using an antibody against phosphorylated cofilin-2. Results: 20 (90.9 %) embryos from Y-27 group and all controls were alive at examination. Morphological abnormalities were detected in 14 (70 %) Y-27 embryos. Somites appeared improperly developed, flattened in the cranio-caudal direction, and elongated in transverse direction in relation to controls. Chick embryos in Y-27 also presented with tortuosity of the neural tube in the lumbosacral region. Western blot analysis showed inhibition of cofilin-2 phosphorylation in affected embryos in comparison to controls. Conclusion: Our study provides evidence that ROCK inhibitor results in the disruption of normal somitogenesis in chick embryos which may contribute to the failure of fusion of the anterior abdominal wall causing VBWD. [ABSTRACT FROM AUTHOR]

Purpose: Although the precise pathogenesis of ventral body wall (VBW) defects is not clearly understood, it has recently postulated that disruption of somite development during early embryogenesis may cause failure of proper VBW formation. The administration of cadmium (Cd) after 60 h of incubation induces omphalocele spectrum in the chick embryo. Previous studies have shown that one of the earliest histological changes seen in this model is abnormal cell death in the somite, occurring at 4 h post treatment (4H). However, the molecular mechanism by which Cd acts in this critical period of embryogenesis still remains unclear. Presenilins are expressed in somites and play an important role in vertebrate development, including somitogenesis and thus VBW formation. We designed this study to test the hypothesis that gene expression levels of presenilin 1 (PSEN1) and presenilin 2 (PSEN2) are downregulated during the critical period of early embryogenesis in the Cd-induced omphalocele in the chick model. Methods: After 60 h of incubation, chick embryos were exposed to either saline or 50 μM cadmium and divided into two groups: control and Cd ( n = 8 at each time point for each group). Real-time RT-PCR was performed to evaluate the relative mRNA expression levels of PSEN1 and PSEN2 in the Cd-induced omphalocele chick model. Differences between two groups at each time point were analysed statistically and the significance was accepted at p < 0.05. Immunofluorescence confocal microscopy was performed to evaluate the protein expression/distribution of presenilins in the somite of chick embryo. Results: The relative mRNA expression levels of PSEN1 and PSEN2 were significantly downregulated in the Cd group at 4H compared with controls ( p < 0.005) (Table). However, there were no significant differences at the other time points. At 4H, immunofluorescence of presenilins (green) was markedly diminished in the Cd-treated embryos, whereas strong immunofluorescence of them was seen in the somite (dermomyotome) in controls (Fig. 1).[Figure not available: see fulltext.] Conclusion: We provide evidence, for the first time, that gene expression of presenilins is downregulated during the narrow window of very early embryogenesis in the Cd chick model. Decreased expression of presenilins may contribute to omphalocele phenotype in Cd chick model, by disrupting somite development. [ABSTRACT FROM AUTHOR]

Purpose: Administration of heavy metal cadmium (Cd) after 60-h incubation induces omphalocele spectrum in the chick embryo. Although previous studies have shown that the earliest detectable histological changes in the chick Cd model occurs commencing at 4-h post-treatment (4H). However, the molecular mechanism by which Cd acts in the critical period of early embryogenesis still remains unclear. Zic3, a Gli superfamily transcription factor, is expressed in somites and plays an important role in vertebrate development, including somitogenesis and thus ventral body wall formation. Gli3 is also expressed in somites and interacts with Zic3 physically and functionally. It has been reported that Gli3 homozygous double mutants display omphalocele. Zic3 mutant mice have also been known to result in omphalocele phenotype. We designed this study to test the hypothesis that Gli3 and Zic3 gene expression is downregulated during the critical period of very early embryogenesis in the Cd-induced omphalocele in the chick model.Methods: After 60-h incubation, chick embryos were exposed to either saline or 50 μM cadmium and divided into two groups: control and Cd (n = 24 for each group). Real-time reverse transcription polymerase chain reaction was performed to evaluate the relative mRNA expression levels of Gli3 and Zic3 in the Cd-induced omphalocele chick model. Differences between the two groups at each time point were analyzed statistically and the significance was accepted at p < 0.05. Immunohistochemistry was also performed to evaluate the expression/distribution of those proteins in chick embryo.Results: The relative mRNA expression level of Gli3 and Zic3 was significantly decreased in the Cd group at 4H when compared with controls (p < 0.05). However, there were no significant differences at the other time points. At 4H, immunoreactivity of GLI3 and ZIC3 was also markedly decreased in Cd-treated embryos, whereas strong expression of them was seen in the somite in controls.Conclusion: We provide evidence, for the first time, that Gli3 and Zic3 gene expression is downregulated during the narrow window of very early embryogenesis in Cd chick model. Disruption of Gli3-Zic3 interaction in the critical period for ventral body wall formation may contribute to omphalocele phenotype in Cd chick model. [ABSTRACT FROM AUTHOR]

In vertebrate somitogenesis, “segmentation clock” genes ( her in zebrafish, hes in mouse, and hairy in chick) show oscillation, synchronized over nearby cells through intercellular interaction. In zebrafish, neighboring cells interact by Delta-Notch signaling to realize synchronization. Under Delta-Notch, however, a cell with a high expression of the segmentation clock gene tends to suppress its expression in adjacent cells, which might produce spatial heterogeneity instead of synchronized oscillation. Here we studied the conditions under which pre-somitic mesoderm cells show synchronized oscillation of gene expression mathematically. We adopted a model that explicitly considers the kinetics of the mRNA and proteins of the segmentation clock gene and cell–cell interaction via Delta-Notch signaling. From statistical study of a model with randomly generated parameters, we revealed how the likelihood that the system generates stable synchronized oscillation depends on the rate of each reaction in the gene–protein kinetics. [ABSTRACT FROM AUTHOR]

The problem of understanding the role of the time factor in embryogenesis is still at the conceptual stage. At the same time, a number of rhythmic processes described in the embryogenesis of animals point to the involvement of a biological clock in this period of ontogenesis. Most of them (biochemical, biophysical, cytological) have been identified during the process of cleavage and have a duration equal to that of a single cleavage division τ0. The current paper considers mainly the development of salmon fish embryos, which, in comparison with the widely used models ( Danio rerio, Orysias latipes), have some extra advantages: they are about twice as big at similar stages and develop 30–40 times slower. The most pronounced and rhythmical process in vertebrates is somitogenesis. The work presents a series of fundamental facts in relation to the temporal and spatial aspects of somitogenesis: 1) the formation of each new somite occurs within a constant time τ S; 2) this time is controlled to a high degree of accuracy; 3) the size of new somites ( l S) along the anterioposterior axis is constant; 4) the temporal factor, in contrast to the spatial, plays the primary role during morphogenesis of somites. The rhythms τ0 and τ S in different species are equal or are multiples of each other, an indication of their common origin. A high degree of synchronism in embryo development from the start of cleavage to the end of somitogenesis is revealed. This proves the existence of constant temporal control of their development during most of the period of embryogenesis. It is proposed that the endogenic rhythms under discussion are responsible for the coordination and integration of multilevel processes in embryogenesis, compensating for the lack of a formed nervous system during this period. [ABSTRACT FROM AUTHOR]

Spondylocostal dysostosis (SCD) is an inherited disorder with abnormal vertebral segmentation that results in extensive hemivertebrae, truncal shortening and abnormally aligned ribs. It arises during embryonic development by a disruption of formation of somites (the precursor tissue of the vertebrae, ribs and associated tendons and muscles). Four genes causing a subset of autosomal recessive forms of this disease have been identified: DLL3 (SCDO1: MIM 277300), MESP2 (SCDO2: MIM 608681), LFNG (SCDO3: MIM609813) and HES7 (SCDO4). These genes are all essential components of the Notch signalling pathway, which has multiple roles in development and disease. Previously, only a single SCD-causative missense mutation was described in HES7. In this study, we have identified two new missense mutations in the HES7 gene in a single family, with only individuals carrying both mutant alleles being affected by SCD. In vitro functional analysis revealed that one of the mutant HES7 proteins was unable to repress gene expression by DNA binding or protein heterodimerization. [ABSTRACT FROM AUTHOR]

Somites are condensations of mesodermal cells that form along the two sides of the neural tube during early vertebrate development. They are one of the first instances of a periodic pattern, and give rise to repeated structures such as the vertebrae. A number of theories for the mechanisms underpinning somite formation have been proposed. For example, in the "clock and wavefront" model (Cooke and Zeeman in J. Theor. Biol. 58:455- 476, 1976), a cellular oscillator coupled to a determination wave progressing along the anterior-posterior axis serves to group cells into a presumptive somite. More recently, a chemical signaling model has been developed and analyzed by Maini and coworkers (Collier et al. in J. Theor. Biol. 207:305-316, 2000; Schnell et al. in C. R. Biol. 325:179- 189, 2002; McInerney et al. in Math. Med. Biol. 21:85-113, 2004), with equations for two chemical regulators with entrained dynamics. One of the chemicals is identified as a somitic factor, which is assumed to translate into a pattern of cellular aggregations via its effect on cell-cell adhesion. Here, the authors propose an extension to this model that includes an explicit equation for an adhesive cell population. They represent cell adhesion via an integral over the sensing region of the cell, based on a model developed previously for adhesion driven cell sorting (Armstrong et al. in J. Theor. Biol. 243:98-113, 2006). The expanded model is able to reproduce the observed pattern of cellular aggregates, but only under certain parameter restrictions. This provides a fuller understanding of the conditions required for the chemical model to be applicable. Moreover, a further extension of the model to include separate subpopulations of cells is able to reproduce the observed differentiation of the somite into separate anterior and posterior halves. [ABSTRACT FROM AUTHOR]

In the basal chordate amphioxus ( Branchiostoma), somites extend the full length of the body. The anteriormost somites segment during the gastrula and neurula stages from dorsolateral grooves of the archenteron. The remaining ones pinch off, one at a time, from the tail bud. These posterior somites appear to be homologous to those of vertebrates, even though the latter pinch off from the anterior end of bands of presomitic mesoderm rather than directly from the tail bud. To gain insights into the evolution of mesodermal segmentation in chordates, we determined the expression of ten genes in nascent amphioxus somites. Five ( Uncx4. 1, NeuroD/ atonal- related, IrxA, Pcdhδ2- 17/ 18, and Hey1) are expressed in stripes in the dorsolateral mesoderm at the gastrula stage and in the tail bud while three ( Paraxis, Lcx, and Axin) are expressed in the posterior mesendoderm at the gastrula and neurula stages and in the tail bud at later stages. Expression of two genes ( Pbx and OligA) suggests roles in the anterior somites that may be unrelated to initial segmentation. Together with previous data, our results indicate that, with the exception that Engrailed is only segmentally expressed in the anterior somites, the genetic mechanisms controlling formation of both the anterior and posterior somites are probably largely identical. Thus, the fundamental pathways for mesodermal segmentation involving Notch–Delta, Wnt/β-catenin, and Fgf signaling were already in place in the common ancestor of amphioxus and vertebrates although budding of somites from bands of presomitic mesoderm exhibiting waves of expression of Notch, Wnt, and Fgf target genes was likely a vertebrate novelty. Given the conservation of segmentation gene expression between amphioxus and vertebrate somites, we propose that the clock mechanism may have been established in the basal chordate, while the wavefront evolved later in the vertebrate lineage. [ABSTRACT FROM AUTHOR]

A variety of Ca2+ signals, in the form of intercellular pulses and waves, have been reported to be associated with the various sequential stages of somitogenesis: from convergent extension and the formation of the paraxial mesoderm; during the patterning of the paraxial mesoderm to establish segmental units; throughout the formation of the morphological boundaries that delineate the segmental units, and finally from within the maturing somites as they undergo subsequent development and differentiation. Due to both the technical challenges presented in imaging intact, developing embryos, and the subtle nature of the Ca2+ transients generated, they have proved to be difficult to visualize. However, a combination of cultured cell preparations and improvements in explant and whole embryo imaging techniques has begun to reveal a new and exciting class of developmental Ca2+ signals. In this chapter, we review the small, but expanding, number of reports in the literature and attempt to identify common characteristics of the somitogenic Ca2+ transients, such as their mode of generation, as well as their spatial and temporal features. This may help to elucidate the significance and function of these intriguing Ca2+ transients and thus integrate them into the complex signaling networks that orchestrate early developmental events. [ABSTRACT FROM AUTHOR]

Somitogenesis is the key developmental step, which divides the vertebrate body axis into segmentally repeated structures. It requires an intricate process of pre-patterning, which is driven by an oscillator mechanism consisting of the Delta–Notch pathway and various hairy- and Enhancer of split-related ( her) genes. The subset of her genes, which are necessary to set up the segmentation clock, reveal a complex scenario of interactions. To understand which her genes are essential core players in this process, we compared the expression patterns of somitogenesis-relevant her genes in zebrafish and medaka ( Oryzias latipes). Most of the respective medaka genes ( Ol-her) are duplicated like what has been shown for zebrafish ( Dr-her) and pufferfish genes ( Fr-her). However, zebrafish genes show some additional copies and significant differences in expression patterns. For the paralogues Dr-her1 and Dr-her11, only one copy exists in the medaka ( Ol-her1/11), which combines the expression patterns found for both zebrafish genes. In contrast to Dr-her5, the medaka orthologue appears to play a role in somitogenesis because it is expressed in the presomitic mesoderm (PSM). PSM expression also suggests a role for both Ol-her13 genes, homologues of mouse Hes6 ( mHes6), in this process, which would be consistent with a conserved mHes6 homologue gear in the segmentation clock exclusively in lower vertebrates. Members of the mHes5 homologue group seem to be involved in somite formation in all vertebrates (e.g. Dr- and Ol-her12), although different paralogues are additionally recruited in zebrafish (e.g. Dr-her15) and medaka (e.g. Ol-her4). We found that the linkage between duplicates is strongly conserved between pufferfish and medaka and less well conserved in zebrafish. Nevertheless, linkage and orientation of several her duplicates are identical in all three species. Therefore, small-scale duplications must have happened before whole genome duplication occurred in a fish ancestor. Expression of multiple stripes in the intermediate PSM, characteristic for the zebrafish orthologues, is absent in all somitogenesis-related her genes of the medaka. In fact, the expression mode of Ol-her1/11 and Ol-her5 indicates dynamism similar to the hairy clock genes in chicken and mouse. This suggests that Danio rerio shows a rather derived clock mode when compared to other fish species and amniotes or that, alternatively, the clock mode evolved independently in zebrafish, medaka and mouse or chicken. [ABSTRACT FROM AUTHOR]

Artificially hatched Seriola species have the problem of malformation, mainly in their vertebrae, head, and mouth parts. To clarify the cause of vertebral malformation, the effects of hypoxia during embryogenesis on the induction of centrum defects was investigated in artificially hatched amberjack Seriola dumerili. Firstly, 7-somite stage embryos were exposed to waters of 0, 12.5, 25, 50, and 100% dissolved oxygen (DO) for 0.5, 1, 2, 3, 4, and 5 h to confirm the effective dose (DO concentration and duration of exposure) of hypoxia that induces somitic disturbances in newly hatched larvae. Exposure of embryos to 12.5% DO concentration for longer than 0.5 h induced somitic disturbances. Following this result, centrum defects in juveniles were investigated by an induction experiment with embryos exposed to 12.5% DO for 2 h at the gastrula, 1- or 2-somite, 10-somite, 15-somite, or heart beating stage. This experiment revealed that centrum defects were induced only during somitogenesis, and somitic disturbances were the premonitory symptom of centrum defects. These results indicate hypoxia during somitogenesis as a possible cause of centrum defects in amberjack. [ABSTRACT FROM AUTHOR]

A novel gene, AmphiSom, was identified in amphioxus Branchiostoma belcheri tsingtauense. Its sequence and developmental expression pattern were determined. AmphiSom transcripts were first detected in the presomitic mesoderm at the late gastrula stage and reached the highest level in the forming and nascent somites in neurulae. However, the expression of AmphiSom was rapidly down-regulated after somites were formed. It was maintained in the most anterior somite and most posterior somite at neurula stages. By 48 h, AmphiSom transcripts were detected only in the developing tail bud but were no longer detected in 72-h larva. Our data demonstrated that the AmphiSom gene is expressed during the development of somites in amphioxus and could play a role in somite formation. [ABSTRACT FROM AUTHOR]

Somitogenesis requires an intricate process of pre-patterning, which is driven by an oscillator mechanism consisting of the Delta-Notch pathway and hairy- (h) and Enhancer of split- [E(spl)] related genes. With the aim of unravelling the complex mechanism of somite pre-patterning, we have conducted an extensive search for h/E(spl)-related genes in the third release of the Danio rerio genomic sequence. We identified 14 new h/E(spl) genes and analysed them by in situ hybridisation for their potential role in the somitogenesis process. We describe here the functional analysis of one of these genes, which we have named her11. her11 is a paralogue of her1 and, similar to her1, is arranged in a head to head fashion with another her gene, namely the previously described her5. It shares an expression in the midbrain-hindbrain boundary with her5, but is in addition cyclically expressed in patterns overlapping those of her1 and her7 and complementary to those of hey1. Furthermore it is expressed in the anterior half of the most caudally formed somites. We show that Delta-Notch pathway genes and fused somites (fss) are necessary for the control of her11 expression. However, some aspects of the her11 regulation suggest that at least one additional as yet unknown gene of the Delta-Notch cascade is required to explain its expression. Morpholino-oligonucleotide-mediated knockdown of her11 shows that it is involved in the zebrafish somitogenesis clock via an interaction with her1 and her7. We have also studied the role of hey1 by morpholino injection, but could not find a direct function for this gene, suggesting that it reflects the output of the clock rather than being a core component of the mechanism. [ABSTRACT FROM AUTHOR]

New data were obtained corroborating that somitogenesis is a rhythmic process, in which the time of somite formation is strictly constant. This constant (τs) can be considered as a natural unit of developmental “biological clock” characterizing rhythmic processes. The constant τs can be determined with an exceptional accuracy that has no analogs among the known biological processes. This fact alone suggests that the accuracy of developmental clock is very high. In addition to the constancy of τs, all forming somites have equal linear size along the notochord axis. In the process with strictly constant temporal and spatial factors, time plays the leading role in triggering the formation of new somite. This became clear in studies of twin embryos. Both embryos had the same number of somites but they were shorter than in the normal embryos. Also, we measured the length of head and both segmented and unsegmented caudal parts of the trunk. Combined with the published data on somitogenesis, our results suggest that the previously proposed scheme for the role of developmental clock in embryogenesis predicts: (1) a possible loss of some embryonic stages without serious consequences for subsequent development and (2) periodic switching on/off any embryonic processes (at the molecular, cellular, or supercellular level) with intervals multiple to τs. [ABSTRACT FROM AUTHOR]

Hairy-related basic helix-loop-helix (bHLH) transcription factors are targets of Delta-Notch signaling and represent essential components for a number of cell fate decisions during vertebrate embryogenesis. Hey genes encode a subfamily of hairy-related proteins that have been implicated in processes like somitogenesis, blood vessel and heart development. We have identified and characterized hey genes in three teleost fish lineages using degenerate PCR and database searches. Phylogenetic analysis of Hey proteins suggests a complex pattern of evolution with high divergence of hey2 in Takifugu rubripes (Fugu, Japanese pufferfish) and possibly loss in the related Tetraodon nigroviridis (the freshwater pufferfish). In addition, duplication of hey1 in both pufferfishes, Fugu and Tetraodon, was observed. Conversely, zebrafish (Danio rerio) has the same complement of three hey genes as known from mammals. All three hey genes show much more restricted gene expression profiles in zebrafish when compared to mouse. Importantly, while all three murine Hey genes are expressed in overlapping patterns in the presomitic mesoderm (PSM) and somites, in zebrafish only hey1 shows PSM and somite expression in a highly dynamic fashion. Therefore, while overlapping expression might account for redundancy of hey function in higher vertebrates, this is unlikely to be the case in zebrafish. In deltaD (dlD) deficient after-eight zebrafish mutants, the dynamic expression of hey1 in the PSM is impaired and completely lost in newly formed somitomeres. Overexpression of dlD on the other hand results in the ectopic expression of hey1 in the axial mesoderm. Hence, hey1 represents a target of Delta-Notch signaling dynamically expressed during somite formation in zebrafish. [ABSTRACT FROM AUTHOR]

It was shown on the example of chick embryo that the number τ0 (τn/τ0) can be recommended as a measure of biological time and, for this purpose, the duration of the minimal mitotic cycle during synchronous cleavage divisions should be determined (in minutes) in various avian species. Based on the preliminary data, one can propose the comparability and similarity of the temporal programs of gastrulation and somitogenesis in the chick embryo and embryos of some fish and amphibians. [ABSTRACT FROM AUTHOR]

A number of genes that are involved in somitogenesis in vertebrates are cyclically expressed in the presomitic mesoderm. These include homologues of the Drosophila genes fringe and hairy. We have analysed here two genes that belong to these classes in the zebrafish, namely the apparent orthologues of lunatic fringe (l-fng) and of c-hairy1 (called her9). However, unlike the respective mouse and chicken genes, they are not expressed cyclically in the presomitic mesoderm. Instead, both genes are mainly expressed in the central nervous system. her9 is predominantly expressed in the fore- and midbrain, and transiently in the hindbrain. Thus, the previously identified and only very distantly related her1 gene of zebrafish has more similarities to the expression of the c-hairy1 gene than its apparent orthologue her9, indicating that sequence similarity and similarity of function are not necessarily linked in this case. l-fng expression is found in alternating pre-rhombomeres, comparable to the equivalent mouse gene expression and in the anterior compartments of the mature somites, which was also shown for the chicken l-fng gene. The latter expression indicates that it might be involved in boundary definition and cell fate decision processes, rather than in pre-patterning of the somites. Interestingly, a similar role has previously been inferred for the grasshopper homologue of l-fng. This suggests that the function of l-fng in boundary definition of the somites might be ancestral, while its recruitment to the pre-patterning process of the somites might be a derived feature in higher vertebrates. [ABSTRACT FROM AUTHOR]

The anatomical and cell biological aspects of somite formation in the chick embryo have been rather well studied. Molecular regulation of somitogenesis in vertebrates is just beginning to be understood. We have studied the effects of human recombinant activin on somitogenesis in gastrulating chick embryos cultured in vitro with a view to assessing the possible role of activin-related molecules in this phenomenon. Activin disrupted somitogenesis in treated embryos, resulting in the formation of abnormal, split or ectopic somites. Light microscopic examination indicated that the ability of activin to interfere with somitogenesis might be partly due to initiation of somite formation at ectopic sites. We show that these cells are indeed somitogenic by their expression of one of the earliest somite-specific marker genes, Pax3. Scanning electron microscopic examination of control and treated embryos revealed direct effects of activin on cell–cell interactions. Cells from treated embryos exhibited disrupted intercellular adhesion leading to large intercellular spaces, altered cell shapes and modification of cell surface protrusions. The effects of activin on somitogenesis appear to be specific, since the neural structures, which are generally more susceptible to chemical insults during gastrulation, were relatively less affected. The results clearly point to a role of activin-related molecules in somitogenesis in the chick embryo. [ABSTRACT FROM AUTHOR]

We studied the roles of β-catenin in somitogenesis using immunostaining and antisense experiments in rat embryos. High levels of β-catenin appeared transiently in the developing rat somites. Initially, β-catenin accumulation was observed in the core cells of presomitic cell aggregates and then in the lumen of epithelial vesicles. Subsequently, it was confined to the dermomyotomes and their lumen and then the myotomes. High levels of cyclin D1 were observed in the core cells, in the lumen of epithelial vesicles, in myotomes, and in mesenchymal sclerotomes. When embryos were cultured in medium supplemented with β-catenin antisense oligodeoxynucleotide (ODN), the accumulation of β-catenin, but not of cyclin D1, in the nascent somites and dermomyotomes was suppressed, while the number of somites was the same as that observed in control embryos. The number of myosin-positive somites and the amount of myosin per somite in embryos treated with the antisense ODN were lower than those in controls. These results suggested that β-catenin promotes development of myotomal cells during somitogenesis. The function of β-catenin in the development of myotomes may not be correlated to cyclin D1. [ABSTRACT FROM AUTHOR]

The eyes absent-like genes encode a group of putative transcriptional coactivators with a sole representative in Drosophila and several members in mammals. Haploinsufficiency of the human EYA1 gene results in branchio-oto-renal syndrome characterized by developmental anomalies of the branchial arches, the three compartments of the ear and the kidney. As a first step towards a functional analysis of this gene in lower vertebrates, we isolated its zebrafish homologue, eya1, and studied its expression pattern during embryogenesis. The eya1 cDNA predicts a protein with 84.7% identity with the human homologue. Transcripts are first detected at the tailbud stage in presumptive cranial placodal precursor cells. Thereafter, eya1 expression continues in anterior pituitary, olfactory, otic, and lateral line placodes. Aside from these placodal sites of expression, eya1 transcripts were observed in the somites, developing pectoral fins, and branchial arches. No expression was found in pronephros or Wolffian duct of the zebrafish renal system. Within the developing ear, eya1 expression becomes confined to the ventral part of the otic vesicle from where the acoustic ganglion precursor cells arise and the sensory patches differentiate. In the lateral line, eya1 is expressed in the placodes, ganglia, migrating primordia, and receptive organs at all developmental stages, including both the differentiating hair and supporting cells. Taken together, these results indicate a remarkable similarity in both the structure and expression pattern of eya1 between higher and lower vertebrates, suggesting that the function of this gene has been conserved throughout vertebrate evolution. [ABSTRACT FROM AUTHOR]

Notch genes encode transmembrane receptors that interact with numerous signal transduction pathways and are essential for animal development. To facilitate analysis of vertebrate Notch gene function, we isolated cDNA fragments of three novel Notch genes from zebrafish ( Danio rerio), Notch1b, Notch5 and Notch6. Notch1b is a second zebrafish Notch1 gene. From analysis of the Notch1b sequence we argue that the various vertebrate Notch gene subfamilies encode receptors with different signalling specificities. Notch5 and Notch6 represent novel vertebrate Notch gene subfamilies. Remarkably, Notch1b lacks expression in presomitic mesoderm, Notch5 is expressed in a metameric pattern within the presomitic mesoderm whilst Notch6 expression is excluded from the nervous system. The expression patterns of these genes suggest important roles in gastrulation, somitogenesis, tail bud extension, myogenesis, heart development and neurogenesis. We discuss the implications of our observations for Notch gene evolution and function. [ABSTRACT FROM AUTHOR]

Somite formation in the mouse embryo begins with the recruitment of mesenchymal cells into the paraxial mesoderm. Cells destined for the paraxial mesoderm are recruited from a progenitor population found first in the embryonic ectoderm and later in the primitive streak and the tail bud. Experimental evidence suggests that the allocation of precursor cells to different mesodermal lineages may be related to the site at which the cells ingress through the primitive streak. An increasing number of genes, such as those encoding growth factor and transcription factors, are now known to be expressed in the primitive streak. It is not known whether the specification of mesodermal cell fate has any relationship with the activity of genes that are expressed in the restricted cell populations of the primitive streak. Somitomeres, which are spherical clusters of mesenchymal cells in the presomitic mesoderm, presage the segmentation of somites in the paraxial mesoderm. The somitomeric organization denotes a pre-pattern of segmentation that defines the physical boundary and the bilateral symmetry of the mesodermal segments in the body axis. The establishment of new somitomeres seems to require the interaction of a resident cell population in the presomitic mesoderm and the incoming primitive streak cells. Cell mixing, which occurs in the somitomeres prior to somite segmentation, poses problems in understanding the developmental role of the somitomere and the real significance of the partitioning of the node-derived and primitive streak-derived cells in the mesodermal segments. In the presomitic mesoderm, the expression of some genes that encode transcription factors, growth factors or tyrosine kinase receptor, and the localization of certain cell adhesion molecules are closely associated with distinct morphogenetic events, such as cell clustering in the presomitic mesoderm and the formation of epithelial somites. There is, however, very little direct relationship between the spatial pattern of gene expression and the somitomeric organization in the presomitic mesoderm. Results of somite transplantation experiments suggest that both the segmental address and the morphogenetic characteristics of the somite may be determined during somite segmentation. Regional identity of the paraxial mesodermal segment is conferred by the expression of a combination of Hox genes in the sclerotome and probably other lineage-specific genes that are subject to imprinting. Superimposed on the global metameric pattern, two orthogonal polarities of cell differentiation are endowed in each mesodermal segment. The rostro-caudal polarity is established prior to somite segmentation. This polarity is later manifested by the subdivision of the sclerotome and the alliance of the neural crest cells and motor axons with the rostral half-somite. The dorso-ventral polarity is installed after somite segmentation, probably by an inductive mechanism similar to that which specifies the dorso-ventral pattern of neuronal differentiation in the neural tube. The segmental pattern of the paraxial mesoderm has been found to exert a direct impact not only on the morphogenesis of the vertebral column, but also on the segmental organization of the lateral mesoderm, the peripheral nervous system and the craniofacial structures. [ABSTRACT FROM AUTHOR]

This is a study on the ability of mouse embryos to compensate for a loss of cells and to develop with body parts of normal size and normal proportions during postimplantation development. Micro-manipulations were performed on 4-cell pre-implantation mouse embryos to reduce the number of cells by 25% (3/4 embryos) or 50% (2/4 embryos). Blastocysts developed from these embryos showed a preferential loss of inner cell mass population, and fewer of them formed viable embryos after implantation. The size of post-implantation 3/4 embryos was initially smaller than controls of the same gestational age, but compensatory growth, achieved by increasing cell numbers at above the normal rate and beyond the normal duration, took place between 6.5 and 11.5 days, resulting in a complete restoration of body size. During compensatory growth the 3/4 embryos rescheduled events of gastrulation and morphogenesis in keeping with cell number or body size appropriate for each developmental stage. The formation of the correct number of somites was accomplished by changing the rate of somite segmentation and by an adjustment of the size of individual somites and somitomeres proportional to the available amount of precursor tissues. Morphogenesis and pattern formation in embryos recovering from earlier cell losses are therefore regulated in accordance to tissue volume (or cell number) instead of chronological age or some intrinsic cellular clock. [ABSTRACT FROM AUTHOR]

It has been suggested that matrix molecules like fibronectin and laminin influence the differentiation and migration of embryonic cells. We investigated the role of these two glycoproteins in somitogenesis as well as in the differentiation and migration of the avian Wolffian (pronephric and mesonephric) duct. At first, we described essential steps in the development of these two organ anlagen by light microscopy, SEM and TEM. To localize fibronectin and laminin more exactly in the actual stages, we used the indirect immunoperoxidase reaction at the light microscopic level and the peroxidase-antiperoxidase technique at the ultrastructural level. Fibronectin was found at the surface of the unsegmented paraxial mesoderm, increasing in the cranial direction, and in the basal laminae of somites and Wolffian duct. The mesenchymal tip of the duct contains a moderate amount of fibronectin. In the two investigated organ anlagen, laminin was found mainly in the basal laminae. The role of fibronectin and laminin was investigated further by using synthetic peptides that mimic the main cell binding domain of either fibronectin or laminin, and that competitively inhibit their cell surface receptors. Thus, the pentapeptides GRGDS, YIGSR, and for control, SHLVE were micro-injected under the ectoderm of 2-day-old embryos. After treatment with GRDS, the Wolffian duct and the segmental plate are more compact. The rounded cells exhibit only short processes and narrow intercellular spaces. At the side of injection the duct shows a delay in migration. After treatment with YIGSR the Wolffian duct migrated laterally over the somatopleure. The basal laminae seem to be incomplete. SHLVE had no effect. Our results suggest that fibronectin is a prerequisite for the migration of the Wolffian duct, and that laminin probably plays a role in guiding the duct. The epithelialization during somitogenesis and differentiation of the duct is a more complex process involving also fibronectin and laminin. [ABSTRACT FROM AUTHOR]

The vertebrate spinal cord shows a segmental pattern of dorsal and ventral nerve roots. It is believed that this peripheral neural segmentation depends on the segmentation of the somitic mesoderm into somites. I have studied the relationship between somite segmentation and the pattern of Rohon-Beard sensory neurite outgrowth from the spinal cord in Xenopus embryos. Most Rohon-Beard neurites grow out in fascicles from the spinal cord at the intersomite clefts, a smaller number of neurites growing out over the dorsal somite margins. Previous work on peripheral neural segmentation has relied on transplanting or destroying somitic mesoderm. Here a non-invasive technique has been used. The pattern of somite segmentation was disrupted by heat shocks, leading to regions of somite fusion where one or two intersomite clefts are absent. At the regions of somite fusion the number of sensory fascicles is then unchanged but their distribution is different. The segmental pattern of sensory fascicle outgrowth is replaced by a fairly even distribution of outgrowth fascicles. Two interpretations of the difference in fascicle outgrowth at fused and control somites are discussed. Firstly, that heat shocks have a direct affect on fascicle outgrowth. Secondly, that heat shocks affect fascicle outgrowth indirectly by disrupting somite segmentation. [ABSTRACT FROM AUTHOR]

This paper suggests that chick somites form because presomitic cells exert tractional forces on one another. These forces derive from the increase in cell adhesion and density that occurs as N-CAM and N-cadherin are laid down by the motile cells of the presomitic mesoderm, well before the somites form. Harris et al. (1984) have shown that adhesive and motile cells in an appropriate environment in vitro can spontaneously form aggregates under the influence of the tractional forces that they exert. Presomitic mesodermal cells may behave similarly: as CAM production increases local adhesivity, the tractional forces between the cells should become sufficiently strong for groups of cells to segment off the mesenchyme as somites. The successive expression of CAMs down the presomitic mesoderm will thus lead to the formation of an anterior-posterior sequence of somites. This mechanism can explain several aspects of somitogenesis that models generating a repetitive pre-pattern through gating cohorts of cells find hard to explain: first, mesodermal segregation occurs among highly adherent cells; second, that multiple rows of somites can form in embryos cultured on highly adherent substrata; third, that stirred mesoderm will still form normal somites; and, fourth, how somite size can be altered in heat-shocked embryos and elsewhere. Suggestions are given as to how the mechanism may be tested and where else in the embryo it could apply. [ABSTRACT FROM AUTHOR]

The spatio-temporal pattern of expression for the Eph receptor tyrosine kinase, Cek8, was observed in normal chick embryos from H–H stage 6 to 23 by immunohistochemical techniques. Expression of Cek8 was already present in embryos at H–H stage 6, where it was located in the neural plate of the brain region, paraxial mesoderm, and the primitive streak. Regions expressing Cek8 subsequently increased during development to include the neural folds of the brain, rhombomeres 3 and 5, the caudal part of the neural plate, neural creast cells related to the formation of glossopharyngeal nerve ganglia, invaginated cells throughout the primitive groove and the epithelium of the rudiment of the gall bladder. Cek8 was also expressed in the mesenchymal cells of the pharyngeal arches, allantoic stalk and limb buds as well as in the areas surrounding the eye vesicles and nasal pits. Furthermore, cells in the tail bud progressing to the secondary neurulation expressed Cek8. Thus, the spatio-temporal patterns of Cek8 expression appears to have intimate relationships with tissue rebuilding, the maturation of differentiated cells, and the spatial organization of tissues. Consequently, it appears that Cek8 plays an integral role in the developmental events leading to the formation of a wide – though not inclusive – variety of tissues and organ systems. [ABSTRACT FROM AUTHOR]

The segmental arrangement of spinal nerves in higher vertebrate embryos provides a simple system in which to study the factors that influence axon pathfinding. Developing motor and sensory axons are intimately associated with surrounding tissues that direct axon guidance. We argue that two distinct guidance mechanisms, viz. contact repulsion and chemorepulsion, act simultaneously to prescribe spinal axon trajectories by ’surround-repulsion’. Motor and sensory axons grow freely within the anterior half of each mesodermal somite, because they are excluded from posterior half-somites by contact repulsion. By contrast, the dorsoventral trajectory that bipolar sensory axons of the dorsal root ganglia follow is governed by diffusible repellents originating from the notochord medially and dermamyotome laterally. Even though spinal nerve development appears to be a simple system for elucidating axon guidance mechanisms, many distinct candidate guidance molecules have been implicated and their relative contributions remain to be evaluated. [ABSTRACT FROM AUTHOR]

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