The segmentation clock: converting embryonic time into spatial pattern

In most animal species, the anteroposterior body axis is generated by the formation of repeated structures called segments. In vertebrate segmentation, a specialized mesodermal structure called the somite gives rise to skeletal muscles, vertebrae, and some dermis. Formation of the somites is a rhythmic process that involves an oscillator--the segmentation clock--driven by Wnt and Notch signaling. The clock ticks in somite precursors and halts when they reach a specific maturation stage defined as the wavefront, established by fibroblast growth factor and Wnt signaling. This process converts the temporal oscillations into the periodic spatial pattern of somite boundaries. The study of somite development provides insights into the spatiotemporal integration of signaling systems in the vertebrate embryo.     

Cell dynamics during somite boundary formation revealed by time-lapse analysis

We follow somite segmentation in living chick embryos and find that the shaping process is not a simple periodic slicing of tissue blocks but a much more carefully choreographed separation in which the somite pulls apart from the segmental plate. Cells move across the presumptive somite boundary and violate gene expression boundaries thought to correlate with the site of the somite boundary. Similarly, cells do not appear to be preassigned to a given somite as they leave the node. The results offer a detailed picture of somite shaping and provide a spatiotemporal framework for linking gene expression with cell movements.     

Hyaluronate in vasculogenesis

Limb buds of chicken embryos contain within the peripheral mesoderm an avascular zone that is rich in hyaluronic acid. Epithelial tissues that synthesize large amounts of hyaluronic acid relative to other glycosaminoglycans caused avascularity when implanted into normally vascular wing mesoderm. Epithelia that synthesize little hyaluronic acid did not cause avascularity. Elvax implants containing hyaluronic acid caused the formation of avascular zones, whereas similar implants containing other glycosaminoglycans did not give rise to avascular zones. Hyaluronic acid may thus play a role in determining the location of blood vessels in the embryo.     

Chemokine signaling controls endodermal migration during zebrafish gastrulation

Directed cell movements during gastrulation establish the germ layers of the vertebrate embryo and coordinate their contributions to different tissues and organs. Anterior migration of the mesoderm and endoderm has largely been interpreted to result from epiboly and convergent-extension movements that drive body elongation. We show that the chemokine Cxcl12b and its receptor Cxcr4a restrict anterior migration of the endoderm during zebrafish gastrulation, thereby coordinating its movements with those of the mesoderm. Depletion of either gene product causes disruption of integrin-dependent cell adhesion, resulting in separation of the endoderm from the mesoderm; the endoderm then migrates farther anteriorly than it normally would, resulting in bilateral duplication of endodermal organs. This process may have relevance to human gastrointestinal bifurcations and other organ defects.     

Synchrony dynamics during initiation, failure, and rescue of the segmentation clock

The "segmentation clock" is thought to coordinate sequential segmentation of the body axis in vertebrate embryos. This clock comprises a multicellular genetic network of synchronized oscillators, coupled by intercellular Delta-Notch signaling. How this synchrony is established and how its loss determines the position of segmentation defects in Delta and Notch mutants are unknown. We analyzed the clock's synchrony dynamics by varying strength and timing of Notch coupling in zebra-fish embryos with techniques for quantitative perturbation of gene function. We developed a physical theory based on coupled phase oscillators explaining the observed onset and rescue of segmentation defects, the clock's robustness against developmental noise, and a critical point beyond which synchrony decays. We conclude that synchrony among these genetic oscillators can be established by simultaneous initiation and self-organization and that the segmentation defect position is determined by the difference between coupling strength and noise.     

A segmentation clock with two-segment periodicity in insects

Vertebrate segmentation relies on a mechanism characterized by oscillating gene expression. Whether this mechanism is used by other segmented animals has been controversial. Rigorous proof of cyclic expression during arthropod segmentation has been lacking. We find that the segmentation gene odd-skipped (Tc-odd) oscillates with a two-segment periodicity in the beetle Tribolium castaneum. By bisecting embryos and culturing the two halves over different time intervals, we demonstrate that Tc-odd cycles with a period of about 95 minutes at 30°C. Using live imaging and cell tracking in green fluorescent protein-expressing embryos, we can exclude that cell movements explain this dynamic expression. Our results show that molecular oscillators represent a common feature of segmentation in divergent animals and help reconcile the contrasting paradigms of insect and vertebrate segmentation.     

视黄酸诱导动物再生研究进展

综述了视黄酸的概念及其衍生物的特征,探讨了视黄酸对脊椎动物再生作用模式和"超再生"的影响,分析了视黄酸对水螅、蚯蚓、扁形动物和节肢动物等无脊椎动物再生能力的影响,并进一步分析了视黄酸对无脊椎动物发育和再生模式的作用。     

From signals to patterns: space, time, and mathematics in developmental biology

We now have a wealth of information about the molecular signals that act on cells in embryos, but how do the control systems based on these signals generate pattern and govern the timing of developmental events? Here, I discuss four examples to show how mathematical modeling and quantitative experimentation can give some useful answers. The examples concern the Bicoid gradient in the early Drosophila embryo, the dorsoventral patterning of a frog embryo by bone morphogenetic protein signals, the auxin-mediated patterning of plant meristems, and the Notch-dependent somite segmentation clock.     

MicroRNA expression in zebrafish embryonic development

MicroRNAs (miRNAs) are small noncoding RNAs, about 21 nucleotides in length, that can regulate gene expression by base-pairing to partially complementary mRNAs. Regulation by miRNAs can play essential roles in embryonic development. We determined the temporal and spatial expression patterns of 115 conserved vertebrate miRNAs in zebrafish embryos by microarrays and by in situ hybridizations, using locked-nucleic acid-modified oligonucleotide probes. Most miRNAs were expressed in a highly tissue-specific manner during segmentation and later stages, but not early in development, which suggests that their role is not in tissue fate establishment but in differentiation or maintenance of tissue identity.     

Advanced cardiac morphogenesis does not require heart tube fusion

The bilateral cardiac mesoderm migrates from the lateral region of the embryo to the ventral midline, where it fuses to form the primitive heart tube. It is generally accepted that migration and fusion are essential for subsequent stages of cardiac morphogenesis. We present evidence that, in Foxp4 mutant embryonic mice, each bilateral heart-forming region is capable of developing into a highly differentiated four-chambered mammalian heart in the absence of midline fusion. These data demonstrate that left-right chamber specification, cardiac looping, septation, cardiac myocyte differentiation, and endocardial cushion formation are preprogrammed in the precardiac mesoderm and do not require midline positional identity or heart tube fusion.     

Retinoic acid signaling restricts the cardiac progenitor pool

Organogenesis begins with specification of a progenitor cell population, the size of which provides a foundation for the organ's final dimensions. Here, we present a new mechanism for regulating the number of progenitor cells by limiting their density within a competent region. We demonstrate that retinoic acid signaling restricts cardiac specification in the zebrafish embryo. Reduction of retinoic acid signaling causes formation of an excess of cardiomyocytes, via fate transformations that increase cardiac progenitor density within a multipotential zone. Thus, retinoic acid signaling creates a balance between cardiac and noncardiac identities, thereby refining the dimensions of the cardiac progenitor pool.     

斑马鱼胚胎左右不对称发育的研究现状与展望

脊椎动物单从体表特征来看,一般呈两侧对称,但是大多数内脏器官和系统在体内多为不对称分布,而且这种分布方式对于器官或系统功能的行使是必须的。左右不对称发育起始于胚胎时期,打破了机体两侧对称的发育模式,在不同脊椎动物中均很保守。目前利用斑马鱼为模式生物,对胚胎左右不对称发育的研究已经取得了一系列进展,包括临时器官Kupffer’s囊泡的形成、左右不对称发育信号在侧板中胚层中的传递、器官不对称分布等。本文就近年来以斑马鱼为模式生物对胚胎左右不对称发育机制研究的进展进行综述。     

gridlock, an HLH gene required for assembly of the aorta in zebrafish

The first artery and vein of the vertebrate embryo assemble in the trunk by migration and coalescence of angioblasts to form endothelial tubes. The gridlock (grl) mutation in zebrafish selectively perturbs assembly of the artery (the aorta). Here it is shown that grl encodes a basic helix-loop-helix (bHLH) protein belonging to the Hairy/Enhancer of the split family of bHLH proteins. The grl gene is expressed in lateral plate mesoderm before vessel formation, and thereafter in the aorta and not in the vein. These results suggest that the arterial endothelial identity is established even before the onset of blood flow and implicate the grl gene in assignment of vessel-specific cell fate.     

Zebrafish Dpr2 inhibits mesoderm induction by promoting degradation of nodal receptors

Nodal proteins, members of the transforming growth factor-beta (TGFbeta) superfamily, have been identified as key endogenous mesoderm inducers in vertebrates. Precise control of Nodal signaling is essential for normal development of embryos. Here, we report that zebrafish dapper2 (dpr2) is expressed in mesoderm precursors during early embryogenesis and is positively regulated by Nodal signals. In vivo functional studies in zebrafish suggest that Dpr2 suppresses mesoderm induction activities of Nodal signaling. Dpr2 is localized in late endosomes, binds to the TGFbeta receptors ALK5 and ALK4, and accelerates lysosomal degradation of these receptors.     

ACTINOMYCIN D: SPECIFIC INHIBITORY EFFECTS ON THE EXPLANTED CHICK EMBRYO

Chick embryos containing 11 to 13 somites were cultured for 48 hours on media containing various amounts of the antibiotic actinomycin D. Morphological observations as well as quantitative determinations of protein nitrogen, DNA, and RNA indicate that the antibiotic specifically inhibits the growth and development of the embryonic axis posterior to the 12th somite.     

Region-specific neural induction of an engrailed protein by anterior notochord in Xenopus

Anterior-specific neural induction can be assayed by means of an antibody that recognizes the Xenopus homeobox-containing protein En-2. The En-2 antigen is an excellent early marker, since it is present as a discrete band in the anterior neural plate of neurula embryos. Regional induction was assayed by combining dorsal mesoderm with competent ectoderm. Anterior notochord from the early neurula induced En-2 frequently, while posterior notochord induced En-2 less frequently. Presumptive somitic mesoderm and presumptive head mesoderm, though they induced neural tissue, were not strong inducers of En-2. Thus, anterior notochord may be the primary mesodermal tissue responsible for the patterning of the anterior neural plate.