The three-dimensional (3D) microanatomy of the cornea is the major determinant of its optical and mechanical properties. Scanning electron microscopy (SEM) is the most commonly used method to obtain information on the overall 3D microanatomy of organs. However, SEM has not been successful in revealing the 3D microanatomy of the cornea, because the interior of the cornea is too compact to be explored by the electron beam. In this study, the 3D organisation of the cells and extracellular materials of human and rabbit corneas was examined after exposure by HCl and NaOH digestion, and by microdissection by the adhesive tape method. In the cornea of both species, all epithelial cells exhibited microplicae regardless of their location. This raises doubts about the tear film-holding role assigned to the microplicae of the superficial cells. Human and rabbit corneas differed in the collagen fibre patterns of the epithelial basement membranes. The 3D organisation of the stromal lamellae was similar in both species. In humans and rabbits, the keratocytes showed similar 3D features. However, the surface of human keratocytes located near Descemet's membrane exhibited small fenestrations that were not present in the rabbit keratocytes. The pattern of keratocyte innervation by the stromal neural plexus and 3D keratocyte microanatomy confirms that keratocytes form a large intercommunicating network within the corneal stroma. Two morphologically discrete subpopulations of keratocytes located at different stromal levels were identified in both human and rabbit corneas, suggesting that keratocytes are not functionally homogeneous. In addition, the density of the stromal neural plexus appeared to be greater in rabbits than in humans. Clear differences between human and rabbit corneas were observed in the collagen arrangement in Descemet's membrane, which may reflect their different biomechanical requirements.
The first rudiment of the central nervous system is a simple tube, the neural tube, and its cavities become the cerebro-ventricular system. The elements located within this system, their composition and precise morphogenetic role are poorly understood. This study used transmission (TEM) and scanning (SEM) electron microscopy, and immunoelectron microscopy, and describes in the chick the development, ultrastructure, composition, and regression of a previously undescribed extracellular structure located in close relationship with the luminal pole of the developing rhombencephalic tectoria lamina. We have called it the rhombencephalic roof network (RRN). The RRN was first observed in stage 12, closely related to a cluster of apoptotic cells. Between stages 15 and 18, the RRN attained its greatest development; it was rhomboid in shape and SEM revealed a network of fibers. Between stages 19 and 22, the RRN underwent a process of fragmentation and regression, and was not observed after stage 23. With TEM, the RRN appeared formed by amorphous ruthenium-red-positive material and sets of tubes between 4 and 25 nm in diameter. Each tube was formed by the superposition of annular units. Immunolabelling showed the presence of laminin and heparan sulfate proteoglycan in both the amorphous material and fibers; the former also contained tenascin. In terms of ultrastructure and composition, the fibers were similar to one the basic components of the lamina densa of basement membranes. The developing tectoria lamina exhibited openings as early as stage 12+, showing that the neural cavity is not a closed system and that the neural tube fluid (NTF) could be a circulating liquid. The presence in the RRN of three molecules of the extracellular materials actively involved in several developmental processes and the very early appearance of the RRN suggest that this structure plays a developmental role in rhombencephalic morphogenesis.
Supraependymal cellular elements are a constant feature in the adult cerebroventricular system. However, there has been no analysis of their distribution and morphology during the embryonic stages of the chick brain. The ultrastructural features of the rhombencephalic luminal surface of chick embryos ranging from stage 10 to 22 were studied with both scanning and transmission electron microscopy. In addition, immunocytochemistry and confocal laser microscopy were used to examine the presence of 68 kD neurofilaments in supraependymal elements. The ultrastructural observations revealed significant morphological differences in the apical cell surface between the cells at rhombomere boundaries and those in the rhombomere bodies. These differences support the idea that the boundary and the body of rhombomeres contain two morphologically distinct cell types. Supraependymal (SE) cells and SE fibers were present in the rhombencephalon of all embryos studied from stage 12 to 22. The cells were bipolar spindle-shaped. The SE fibers showed a characteristic spatial pattern within the rhombencephalon, following a straight course parallel to the rhombomere boundaries. The SE fibers showed varicosities and their endings contained small vesicles. Both SE cells and SE fibers were positive for 68 kD neurofilaments. Their morphology and reactivity for neurofilaments indicate a neuronal function. The constant presence of SE cells and SE fibers on the surface of the developing rhombencephalon, their special pattern and close relationship with the neural tube fluid (NTF) suggest that these supraependymal elements may be involved in a neuronal signalling pathway between different parts of the same rhombomere and also in chemical communication and integration within the ventricular system, linking distant parts of the developing central nervous system by means of NTF.
Extracellular material molecules play a key role in the regulation of morphogenesis and differentiation of a large number of organs including the central nervous system. However, the role of the neural basement membrane in the growth of different parts of the neural tube has yet to been delineated. Here, the structural and compositional modifications of the basement membrane (BM) of rhombencephalic tectoria lamina anlage (RTLA) have been examined during the process of RTLA epithelial attenuation. Between stages 10 to 11-the presumptive RTLA epithelium showed a structure, thickness and cell-proliferating capacity similar to those observed in other zones of the rhombencephalic walls. Moreover, the rhombencephalic vesicles were surrounded by a continuous BM that was heterogeneous both ultrastructurally and with regard to ruthenium red, laminin and tenascin distribution. After stage 11, the RTLA epithelium underwent a rapid process of attenuation and change to a stratified flattened epithelium. During this remodelling process, apoptosis and inhibition of both PCNA expression and 3H-thymidine uptake occurred in the RTLA epithelium. The BM of the RTLA underwent a process of degration at the beginning of the remodelling, and apoptosis and cell proliferation inhibition of RTLA epithelium were also observed. The loss of the biochemical signals encoded within the BM could lead to cell shape changes, cell proliferation inhibition and to the anoikis type of cell death. Our findings support the idea that the BM surrounding the neural tube plays a key role in controlling both the structure and growth of the CNS during the early developmental stages.
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.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.