Cardiac myocytes are the first cells to differentiate during the development of a vertebrate embryo. A wide variety of molecules take part in various steps in this process. While exploring biologically active molecules from marine sources, we found that a constituent of perivitelline fluid from embryos of the Indian horseshoe crab can enhance growth and differentiation of chick embryonic heart. We have purified the factor and identified the cardiac promoting molecule to be a novel lectin. We show that this molecule influences cardiac development by increasing the number of cells constituting the heart and by modulating the expression of several cardiac development regulatory genes in chick embryos. Using mouse embryonic stem cells we show that the cardiac myocyte enhancing capacity of this molecule extends to mammals and its effects can be blocked using methylated sugars. This molecule may prove to be an important tool in the study of cardiomyocyte differentiation.
In the present study, we show that insulin accelerates early morphogenesis in gastrulating chick embryo explants cultured in vitro, whereas antiserum to insulin adversely affects this process. Comparison between length of body axis of control and treated embryos clearly brings out the significant acceleration of development by excess insulin (0.175 to 17.5 nM). In embryos treated with 87.5 and 175 nM insulin, a high occurrence of abnormalities is observed. Treatment of embryos with antiserum to porcine insulin results in a high percentage of abnormalities, particularly in the forming neural tube. In situ hybridization of whole embryos using digoxigenin-labeled riboprobes showed that insulin modifies the expression of crucial developmental genes within 2 hours. While Brachyury, a pan-mesodermal marker gene, ERNI, the earliest known marker for neural induction in chick, and noggin, important in neural tube patterning, are upregulated, expression of goosecoid, necessary for gastrulation movements, does not appear to be significantly altered. During the same time, insulin does not exert any mitogenic effect on chick embryonic cells as assessed by nuclear counts. These findings demonstrate that insulin plays an important role in the early morphogenesis of the chick embryo. The function of insulin appears to be mediated by specific genes which orchestrate pattern formation during early development.
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.
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