Embryos and larvae of the starfish Pisaster ochraceus are surrounded by a complex extracellular matrix (ECM) layer called the hyaline layer (HL). A similar but less well-organized ECM layer lines some regions of the larval gut. Examination of material preserved by freeze substitution shows that the HL consists of a coarse outer meshwork, a boundary layer, a supporting layer, which is divided into three sublayers, H1, H2, and H3, and an intervillus layer. The development of the HL has been studied in material preserved by freeze substitution. Development begins at fertilization when exocytosis of the cortical granules releases ECM into the perivitelline space and elevates the fertilization membrane. Shortly after, plaques of dense material with attached fibers are present on the outer surface of the egg plasmalemma. Following this, these plaques and fibers are associated with the tips of short microvilli, suggesting that they may induce microvillus formation. Next, the tips of some of the microvilli are joined by short regions of the H1 sublayer. Some of these H1 regions have short segments of boundary layer material associated with their outer surfaces while others are naked. Just prior to hatching, the H1 and boundary layers completely surround the embryo, separating the developing coarse meshwork and intervillus layers. Short segments of the H2 and H3 sublayers are also present. Posthatching, the microvilli and all HL layers increase in thickness and density, particularly the H2, boundary, and coarse outer meshwork layers. The results suggest a sequential organization of HL components from ECM that is secreted into the perivitelline space.
The hyaline layer (HL) is a multilayered extracellular matrix (ECM) that coats the external surfaces of sea urchin and starfish embryos. It is thought to protect and lubricate the embryo, stabilize the blastomeres during morphogenesis, and regulate nutrient intake. Ultrastructural studies of chemically fixed embryos have shown the HL to consist of two to four sublayers. However, since chemical fixatives may cause collapse and alter the positions and antigenicity of the extracellular components, fixation methods that exclude chemicals may reveal a picture of the HL closer to what is present in vivo. Freeze substitution, a fixation method whereby tissues are rapidly frozen and dehydrated at low temperatures, has proved useful for fixing material rich in ECM. In this study, embryos of the starfish Pisaster ochraceus were fixed for microscopy using freeze substitution and three chemical methods in order to determine, as accurately as possible, the structure of the HL. Embryos appear to be best preserved by freeze substitution and demonstrate a HL consisting of at least six distinct sublayers. Based on staining with anionic dyes, most sublayers appear to contain glycosaminoglycans. Freeze substituted embryos, which were also stained with monoclonal antibodies raised against their ECM, revealed that some molecules are common to all six sublayers, whereas other molecules may be restricted to specific sublayers. This suggests that each sublayer could have a different function. Additional evidence suggests that microvillus associated bodies, present in other marine invertebrate embryos, may anchor the asteroid HL to the cell surface microvilli.
This patient demonstrates the truly dynamic nature of CCMs and the increased incidence of new lesions in the setting of CCM1 mutation. This case is remarkable not only for the unprecedented rate of lesion formation (approximately 22 per yr), but also because of the nearly unilateral distribution of the lesions.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.