Prenatal developmental sequences of the esophageal epithelium in the New Zealand white rabbits: Light and electron microscopic analysis.

Several morphogenetic sequences occur during esophageal development and birth defects occur due to defects in foregut morphogenesis. This work aimed to record the cellular events in the morphogenesis of rabbits' esophageal epithelium. On the 16th day of gestation, the esophageal epithelium varied from stratified ciliated columnar to stratified squamous type. The surface epithelium presented mucous cells with mucigen granules of various sizes occupying their supranuclear cytoplasm. Cytoplasmic vacuolation was evident in all layers of the esophageal epithelium at this age. On the 18th gestational day, some light cells could be detected in the middle portion of the epithelium, while others occupied the whole epithelial length. On the 21st day, mucous cells are more frequently observed at the apical esophageal part as well as at the surface epithelium. Numerous elongated dark cells could be distinguished embedded between the basal cells. On the 24th gestational day the number of the mucous cells reached its peak. Reaching the 30th gestational day, several lamellar bodies, a keratinized layer and mitotic divisions could be demonstrated, and the number of both mucous and dark cells was greatly decreased. Collectively, detection of surface mucous and dark cells together with the non-cornified surface in some regions of the rabbit esophageal epithelium at the end of gestation ensure a postnatal development to reach the adult epithelium essential to sustain the passage of the harsh raw food. Future immunohistochemical studies are recommended to investigate the components of secretions in mucous cells and functional studies to highlight the dark cells significance. RESEARCH HIGHLIGHTS: Esophageal epithelium of fetal rabbit was analyzed by light and transmission microscopy. Surface epithelium presented mucous cells with mucigen granules of various sizes. They reached their maximum number on 24th day then decreased. On the 16th day, cytoplasmic vacuolation was evident in all epithelial layers. On the 21st day, numerous elongated dark cells could be distinguished embedded between the basal cells. Before birth, several lamellar bodies, a keratinized layer and mitotic divisions could be demonstrated, and the number of both mucous and dark cells was greatly decreased.

The ultrastructural features of the infundibulum of the green iguana, Iguana iguana.

The purpose of this study is to describe, in detail, the ultrastructure of the infundibulum of the sexually mature and active female green iguana, Iguana iguana. The infundibulum of five iguanas was remarkably distinct from the uterus, and was also clearly demarcated into cranial (expanded v-shaped) and caudal (tubular) divisions. Tissue samples obtained from five portions (three from the cranial division and two from the caudal division) of the infundibulum were processed conventionally for light and electron microscopy. The epithelial lining of the most anterior, middle, and posterior, parts of the cranial division displayed nonciliated cells predominantly, and occasionally ciliated cells. The numerous secretory granules in nonciliated type 1 cell found in the fimbrial aspect of the infundibulum were homogenous and deeply electron-dense, but those in the other two regions were variants of this cell type because they contained variably electron-dense secretory granules. Two main types of nonciliated cells (type 2 and its variant, type 3, as well as type 4) occurred in the epithelial lining of the caudal division of the infundibulum, but they, clearly, showed no dense secretory granules. Whereas the nonciliated type 2 cell and its variant (type 3 cell) contained large glycogen deposits, the type 4 cell lacked these deposits but its apical part contained large lipid-like droplets and, remarkably, blebbed into the duct lumen. The nonciliated cells lining the mucosal tubular glands contained highly electron-dense secretory granules, which were similar to those found in the nonciliated type 1 cell in the epithelial lining of the fimbrial part of the cranial division of the infundibulum.

Ultrastructure of the female reproductive organs of the diving beetle Deronectes moestus incospectus (Leprieur, 1876) (Dytiscidae, Hydroporinae).

We describe the ultrastructure of the female reproductive organs of Deronectes moestus (Dytiscidae Hydroporinae). The long spermathecal duct has a simple epithelium lined internally by a thin cuticle and externally by a thick layer of muscle cells. The wide duct lumen contains electron-dense material, among which remnants of extracellular material are visible. This material consists of tubular structures assembled around sperm bundles previously described in the male deferent ducts. The so-called gland, disposed along the spermathecal duct, is a structure with epithelial cells lined by an irregular cuticle bearing a rich system of microvilli. Many mitochondria are visible in the apical cytoplasm of the epithelial cells, and a few spheroidal bodies are close to the basal nuclei. Since the epithelial ultrastructure of the gland suggests it is involved in fluid uptake from the lumen rather than secretory activity, the term gland, coined by other authors to describe this organ, is inappropriate. The spermatheca is a large structure with a complex epithelium showing secretory and duct-forming cells. The lumen of this organ contains sperm with the distinctive ultrastructural features of those described in the male deferent ducts, namely having a mitochondrial matrix with a small crystallized area and electron-dense dots. Because to its overall organization, the spermatheca of D. moestus can be considered a more integrated organ than those in previously studied hydroporine species.

Capsule-epithelium-fibre unit ultrastructure in the human lens.

This study aimed to investigate the capsule-epithelium-fibre unit ultrastructure of the human lens, particularly the interfaces of the epithelium with the capsule and the epithelium with the fibre cell. A total of 12 lenses from donor humans who died of trauma without systemic and ocular diseases were investigated by transmission electron microscopy (TEM), combined with immunofluorescence staining for localising certain specific proteins. Some of the results were further studied in the anterior lens capsules of cataract patients. Our results revealed capsule protrusion into the epithelium in some areas and potential processing of capsule components. The young elongating fibre cells directly adjacent to the epithelium with a high stain density strongly expressed CD24. Numerous extracellular vesicles could be seen in the space between human lens epithelial cells (HLECs) and between HLECs and the capsule. Mitophagy and autophagy were also observed in the HLECs. Our research may be beneficial in better understanding the function of the human lens.

Preen gland of the laughing dove (Streptopelia senegalensis aegyptiaca): Light and electron microscopic analysis.

This work reviews the microscopic anatomy of the preen gland in laughing dove (Streptopelia senegalensis aegyptiaca) using light, scanning, and transmission electron microscopes. The gland possessed two large pea-shaped lobes. The glandular lobules of each lobe were huddled in elliptical, triangle, round shapes, connected with each other by strands of connective tissue septae. The lobule was composed of glandular follicles, each follicle folded and enclosed by a sheath of connective tissue connected with the neighboring ones by interfollicular septae. The gland's parenchyma was coated with a dense connective tissue capsule composed of collagen, elastic, and reticular fibers. The secretory tubules were divided into peripheral tubules and central tubules. The central ones were located close to the major cavity and lined with thin epithelium, whereas the peripheral tubules were adjacent to the capsule and characterized by taller epithelium. The central secretory tubules were lined with four cell layers: flattened basal, large-sized polyhedral intermediate, and secretory cell layers, as well as a degenerative cells layer that formed of small cells with pale cytoplasm and pyknotic nuclei. Variable sizes and shapes of Herbst corpuscles were detected alongside the papillary duct and near the glandular lobe. Transmission electron microscopy view revealed that the cytoplasm of the intermediate cells contained a dense population of mitochondria, while the secretory and degenerative cells contained fewer mitochondria. In conclusion, these structures will be beneficial for understanding the habitat differences of laughing doves. RESEARCH HIGHLIGHTS: Grossly, the preen gland (PG) of the laughing dove formed of two large pea-shaped lobes. The glandular lobule was composed of glandular follicles, each follicle was folded and enclosed by a sheath of connective tissue connected with the neighboring ones by interfollicular septae. The central secretory tubules were lined with four cell layers: basal, intermediate, secretory, and degenerative cell layers. Variable sizes and shapes of Herbst corpuscles were detected alongside the papillary duct and near the glandular lobe of the PG. In transmission electron microscopic analysis, the cytoplasm of the intermediate cells contained a dense population of mitochondria, while the secretory and degenerative cells contained fewer mitochondria.

Gill histology and ultrastructure in Aplysia depilans (Mollusca, Euopisthobranchia).

The gill of Aplysia depilans consists of several wedge-shaped pinnules with a highly folded structure, differing from the typical ctenidial gills of mollusks. Light microscopy and transmission electron microscopy were used to investigate this organ in juveniles and adults. In this species, the gill epithelium comprised ciliated, unciliated, and secretory cells. The ultrastructural analysis suggests other functions for the gill besides respiration. The deep cell membrane invaginations associated with mitochondria in the basal region of epithelium point to a role in ion regulation. Endocytosis and intracellular digestion were other activities detected in epithelial cells. In juveniles, an intranuclear crystalline structure was seen in some ciliated cells. The presence of an intranuclear crystalline structure was frequently associated with chromatin decondensation, swelling of the nuclear envelope and endoplasmic reticulum cisternae, and abundance of Golgi stacks. As these intranuclear inclusions were not found in the gill of the adult specimens, their occurrence in the two juveniles seems likely to be an anomalous condition whose cause cannot be established at the moment. Mucous cells were the most abundant secretory cells in the epithelium, but a few epithelial serous cells were also found. In addition, large protein-secreting subepithelial cells had the main cell body inserted in the connective tissue and a long thin neck crossing the epithelium. Mucous cells can be considered responsible for the production of the mucus layer that protects the epithelium, but the specific functions of the epithelial and subepithelial protein-secreting cells remain elusive. Below the epithelium, a layer of connective tissue with muscle cells lined the narrow hemolymph space. The connective tissue included cells with a large amount of rough endoplasmic reticulum cisternae. Bacteria were found on the surface of the gill, and the most abundant had a thin stalk for attachment to the epithelial cells.

Pre-gastric secretory epithelium: A light, scanning and transmission electron microscopic study of an epithelial modification of the esophagus in embryonic quails.

The current study investigated epithelial modification of embryonic quail esophagus using gross examination, light microscopy, transmission and scanning electron microscopy. By semithin sections, the pre-gastric modified region had unfolded mucosa, formed epithelial flabs and pockets, and had reduced muscularis mucosae, thin muscular layer, less glandular tissue, and outer esophageal groove. Conversely, the normal esophageal mucosa was folded, had abundant glandular tissue and prominent muscularis mucosae, with two muscular layers; the outer and the inner. The modified epithelium resembled stratified squamous type that had a high affinity for PAS, methylene blue, and PAP stains. Ultra-structural features of the modified esophageal epithelium resembled stratified squamous epithelium and contained hypertrophic Keratinocytes; dark and light. Hypertrophic keratinocytes had RER organized, few ribosomes, and developed loose bundle of cytokeratin compared with squamous keratinocytes. Hypertrophic Keratinocytes synthesize two types of granules; peripherally located small electron-dense granules and large electron-lucent granules. Hypertrophic keratinocytes had peroxisomes that were identified by the crystalline core of the urate oxidase. In conclusion, epithelia modification may have secretory function. Further studies should be carried out to explain the exact function of this type of modified epithelium.

Ultrastructure of the palatine tonsils of the donkey (Equus asinus): New insights by light, scanning, and transmission electron microscopy.

The study aimed to explore the ultrastructure of the donkeys' palatine tonsils. Palatine tonsils of five male donkeys (5 years old) were investigated macroscopically and microscopically. The tonsils appeared as a dome shape with slight elevation and a circular opening on the surface of the oropharynx. The central tonsillar crypt appeared on the medial side of the palate-pharyngeal folds and the floor of the oropharynx. The external surface of the palatine tonsil had different sizes of mucosal folds, some grooves directed to drainage at the tonsillar opening, and the tonsil crypt opening was a crescentic or irregular oval shape. The outer surface was covered by stratified squamous epithelium and modified to be reticular epithelium invaded by lymphocytes in the crypt called lympho-epithelium. The tonsil crypt had aggregated lymphoid nodules, and the cryptal epithelium has surrounded by diffused lymphocytes and hassles corpuscles-like structures. The lymphocytes infiltrated into different layers of the cryptal epithelium and transformed into reticular or lympho-epithelium. The organized lymphoid nodules were primary and secondary, and the secondary ones had a light germinal center. The interfollicular area had many high endothelial venules and blood capillaries. The endothelial venules were lined by simple cuboidal epithelium and had lymphocytes. The blood capillaries had red blood cells and neutrophils. The tonsil was surrounded incompletely by a connective tissue capsule with mucous glands under that capsule. In conclusion, the epithelial lymphocyte infiltration, crypt epithelium, lymphoid nodules, and intra-follicular area of the donkey's palatine tonsils indicate the humoral and cell-mediated immunological process.

More than a simple epithelial layer: multifunctional role of echinoderm coelomic epithelium.

In echinoderms, the coelomic epithelium (CE) is reportedly the source of new circulating cells (coelomocytes) as well as the provider of molecular factors such as immunity-related molecules. However, its overall functions have been scarcely studied in detail. In this work, we used an integrated approach based on both microscopy (light and electron) and proteomic analyses to investigate the arm CE in the starfish Marthasterias glacialis during different physiological conditions (i.e., non-regenerating and/or regenerating). Our results show that CE cells share both ultrastructural and proteomic features with circulating coelomocytes (echinoderm immune cells). Additionally, microscopy and proteomic analyses indicate that CE cells are actively involved in protein synthesis and processing, and membrane trafficking processes such as phagocytosis (particularly of myocytes) and massive secretion phenomena. The latter might provide molecules (e.g., immune factors) and fluids for proper arm growth/regrowth. No stem cell marker was identified and no pre-existing stem cell was observed within the CE. Rather, during regeneration, CE cells undergo dedifferentiation and epithelial-mesenchymal transition to deliver progenitor cells for tissue replacement. Overall, our work underlines that echinoderm CE is not a "simple epithelial lining" and that instead it plays multiple functions which span from immunity-related roles as well as being a source of regeneration-competent cells for arm growth/regrowth.

Ultrastructure of proboscis blood vessels in females of Bonellia viridis (Annelida: Bonellinae): New reconstructions.

Many data on echiurid anatomy and ultrastructure are obtained for Bonellia viridis and extrapolated to other species. The ultrastructure of the axial blood vessels, which has been described as an "osmotic pump," is regarded as one of the unusual features of echiurids. In this study, the ultrastructure of the proboscis blood vessels in females of B. viridis is described, illustrated by accurate schemes, and a new reconstruction of the axial blood vessel is suggested. The walls of the axial and lateral vessels of the proboscis are formed by myoepithelial cells, which are connected to each other via adherence junctions, underlined by basal lamina, and therefore form a true epithelium. Apical, middle, and basal parts of the myoepithelial cells form long, thin projections, which extend to the connective tissue (in axial vessel) or coelomic canals (in lateral vessels) and to the lumen of the vessels. The presence of such projections may evidence active cellular transport. Similarity in the fine structure of the myoepithelial cells of axial and lateral blood vessels evidence their common origin from myoepithelial cells of the coelomic lining. However, in evolution, the coelomic canals were retained around the lateral vessels and disappeared around the axial vessel. The reduction of a hypothetical ancestral axial coelom may be caused by the extensive development of the connective tissue and muscles in the central part of the proboscis, where the axial vessel extends.

High Glucose Reduces the Paracellular Permeability of the Submandibular Gland Epithelium via the MiR-22-3p/Sp1/Claudin Pathway.

Tight junctions (TJs) play an important role in water, ion, and solute transport through the paracellular pathway of epithelial cells; however, their role in diabetes-induced salivary gland dysfunction remains unknown. Here, we found that the TJ proteins claudin-1 and claudin-3 were significantly increased in the submandibular glands (SMGs) of db/db mice and high glucose (HG)-treated human SMGs. HG decreased paracellular permeability and increased claudin-1 and claudin-3 expression in SMG-C6 cells. Knockdown of claudin-1 or claudin-3 reversed the HG-induced decrease in paracellular permeability. MiR-22-3p was significantly downregulated in diabetic SMGs and HG-treated SMG-C6 cells. A miR-22-3p mimic suppressed claudin-1 and claudin-3 expression and abolished the HG-induced increases in claudin-1 and claudin-3 levels in SMG-C6 cells, whereas a miR-22-3p inhibitor produced the opposite effects. Specificity protein-1 (Sp1) was enhanced in diabetic SMGs and HG-treated SMG-C6 cells, which promoted claudin-1 and claudin-3 transcription through binding to the corresponding promoters. A luciferase reporter assay confirmed that miR-22-3p repressed Sp1 by directly targeting the Sp1 mRNA 3'-untranslated region (3'-UTR). Consistently, the miR-22-3p mimic suppressed, whereas the miR-22-3p inhibitor enhanced, the effects of HG on Sp1 expression. Taken together, our results demonstrate a new regulatory pathway through which HG decreases the paracellular permeability of SMG cells by inhibiting miR-22-3p/Sp1-mediated claudin-1 and claudin-3 expression.

Bi-allelic mutations in MCIDAS and CCNO cause human infertility associated with abnormal gamete transport.

Reduced generation of multiple motile cilia (RGMC) and the consequent primary ciliary dyskinesia (PCD) cause infertility due to a substantial reduction in the number of multiciliated cells (MCCs) in the efferent ducts (EDs)/oviducts. MCIDAS acts upstream of CCNO to regulate the biogenesis of basal bodies (BBs); therefore, both genes play a vital role in the multiciliogenesis of the reproductive tract epithelium. In this study, whole-exome sequencing was performed to identify the causative genes in 10 unrelated infertile patients with PCD: seven males and three females. Notably, homozygous frameshift mutations in MCIDAS (c.186dupT, p.Pro63Serfs*22) and CCNO (c.262_263insGGCCC, p.Gln88Argfs*8) were identified in one male and one female participant from two unrelated consanguineous families. Haematoxylin-eosin staining/scanning electron microscopy revealed abnormal MCCs in the mutated EDs/oviducts. Furthermore, transmission electron microscopy revealed significantly reduced BBs. Immunofluorescence staining showed the absence of MCIDAS and CCNO signals in the affected tissues and confirmed that MCIDAS acts upstream of CCNO in the context of multiciliogenesis in the reproductive tract epithelium. In vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) was successful, with a positive pregnancy outcome in both MCIDAS- and CCNO-mutated patients. Our results support the use of IVF/ICSI interventions to treat infertility due to RGMC in couples.

Morphological analysis of interstitial cells in murine epididymis using light microscopy and transmission electron microscopy.

Smooth muscle contraction of the epididymis plays an important role in sperm transport. Although PDGFRα-positive interstitial cells (PDGFRα (+) ICs) are thought to be involved in controlling smooth muscle movement via intercellular signaling, they have not yet been reported to date in the epididymis. Therefore, we aimed to investigate the morphological characteristics of PDGFRα (+) ICs in the interstitial space of the murine epididymis. Immunohistochemistry showed that PDGFRα (+) ICs co-labeled with CD34 (PDGFRα (+) CD34 (+) ICs were distributed in the interstitial space of the murine epididymis from the initial segment (IS) to the cauda of the epididymis. PDGFRα (+) ICs that were not co-labeled with CD34 (PDGFRα (+) CD34 (-) ICs) were observed just beneath the epithelium from the corpus to the cauda but not in the IS. Both types of PDGFRα (+) ICs were in close proximity to each other as well as the surrounding nerves and macrophages. In addition, PDGFRα (+) CD34 (-) ICs beneath the epithelium were also in close proximity to the basal cells. Using transmission electron microscopy, we identified ICs that possessed elongated and woven cellular processes and were in close proximity to each other, surrounding the cells in the interstitial space. In the murine epididymis, it is suggested that there are two subtypes of ICs that show different distribution patterns depending on the segment, which may reflect segmental differences in mechanisms of sperm transport, forming a cellular network by physical interactions in the murine epididymis.

Robustness of epithelial sealing is an emerging property of local ERK feedback driven by cell elimination.

What regulates the spatiotemporal distribution of cell elimination in tissues remains largely unknown. This is particularly relevant for epithelia with high rates of cell elimination where simultaneous death of neighboring cells could impair epithelial sealing. Here, using the Drosophila pupal notum (a single-layer epithelium) and a new optogenetic tool to trigger caspase activation and cell extrusion, we first showed that death of clusters of at least three cells impaired epithelial sealing; yet, such clusters were almost never observed in vivo. Accordingly, statistical analysis and simulations of cell death distribution highlighted a transient and local protective phase occurring near every cell death. This protection is driven by a transient activation of ERK in cells neighboring extruding cells, which inhibits caspase activation and prevents elimination of cells in clusters. This suggests that the robustness of epithelia with high rates of cell elimination is an emerging property of local ERK feedback.

Copper impair autophagy on zebrafish (Danio rerio) gill epithelium.

Copper (Cu) is an essential element for organism's metabolism, being controversially listed as a priority pollutant. Importantly, the toxicity of Cu has been linked to several cell death pathways. Thus, this study aimed to assess if macroautophagic pathways are triggered by Cu in zebrafish gill, the main target of waterborne pollutants. The electron microscopy findings indicated that Cu induced profound impacts on zebrafish gill structure and functions, being this tissue a biomarker sensitive enough to indicate early toxic effects. The findings also support a clear impairment of autophagy, througth the absence of phagossomes and the significant down-regulation mRNA transcript levels of microtubule-associated protein light chain 3 (LC3). The reduction of LC3 levels was often associated to an increase of apoptotic activation, indicating that the inhibition of macroautophagy triggers apoptosis in zebrafish gills. This study highlighted that the autophagic down-regulation might be affected through the activation of other cell death signaling pathway.

Live Imaging of Epithelial-Mesenchymal Transition in Mesoderm Cells of Gastrulating Drosophila Embryos.

Epithelial-mesenchymal transitions (EMTs) drive the generation of cell diversity during both evolution and development. More and more evidence has pointed to a model where EMT is not a binary switch but a reversible process that can be stabilized at intermediate states. Despite our vast knowledge on the signaling pathways that trigger EMT, we know very little about how EMT happens in a step-wise manner. Live imaging of cells that are undergoing EMT in intact, living, animals will provide us valuable insights into how EMT is executed at both the cellular and molecular levels and help us identify and understand the intermediate states. Here, we describe how to image early stages of EMT in the mesoderm cells of live Drosophila melanogaster embryos and how to image contractile myosin that suspends EMT progression.

Coelomocyte replenishment in adult Asterias rubens: the possible ways.

The origin of cells involved in regeneration in echinoderms remains an open question. Replenishment of circulatory coelomocytes-cells of the coelomic cavity in starfish-is an example of physiological regeneration. The coelomic epithelium is considered to be the main source of coelomocytes, but many details of this process remain unclear. This study examined the role of coelomocytes outside circulation, named marginal coelomocytes and small undifferentiated cells of the coelomic epithelium in coelomocyte replenishment in Asterias rubens. A qualitative and quantitative comparison of circulatory and marginal coelomocytes, as well as changes of circulatory coelomocyte concentrations in response to injury at different physiological statuses, was analysed. The presence of cells morphologically similar to coelomocytes in the context of coelomic epithelium was evaluated by electron microscopy. The irregular distribution of small cells on the surface and within the coelomic epithelium was demonstrated and the origin of small undifferentiated cells and large agranulocytes from the coelomic epithelium was suggested. Two events have been proposed to mediate the replenishment of coelomocytes in the coelom: migration of mature coelomocytes of the marginal cell pool and migration of small undifferentiated cells of the coelomic epithelium. The proteomic analysis of circulatory coelomocytes, coelomic epithelial cells and a subpopulation of coelomic epithelial cells, enriched in small undifferentiated cells, revealed proteins that were common and specific for each cell pool. Among these molecules were regulatory proteins, potential participants of regenerative processes.

Planar cell polarity governs the alignment of the nasopharyngeal epithelium in mammals.

Planar cell polarity (PCP) signalling specifies the orientation of epithelial cells and regulates directional beating of motile cilia of multiciliated epithelial cells. Clinically, defects in cilia function are associated with nasopharyngeal symptoms. The polarity of the nasopharyngeal epithelium is poorly understood. Here, we demonstrated PCP in the nasopharyngeal epithelium. Multiciliated cells (MCCs) were uniformly aligned with their long axis parallel to the tissue axis of the nasopharynx (NP). In addition, PCP proteins exhibited an asymmetrical localisation between adjacent cells. Motile cilia were uniformly aligned in the same direction within both individual cells and neighbouring cells, which manifested as cilial polarity in MCCs. Mutation of Vangl2, a mammalian homologue of the Drosophila PCP gene, resulted in significant disruption of the orientation of epithelial cells. Finally, keratin-5-positive basal cells constantly replenished the luminal ciliated cells; the new dynamic ciliated cells were also oriented parallel to the tissue axis. These results indicate a role for the PCP pathway in the uniform orientation of dynamically replenished epithelial cells in the NP.

Structural and ultrastructural studies on the developing vomeronasal sensory epithelium in the grass snake Natrix natrix (Squamata: Colubroidea).

The sensory olfactory epithelium and the vomeronasal sensory epithelium (VSE) are characterized by continuous turnover of the receptor cells during postnatal life and are capable of regeneration after injury. The VSE, like the entire vomeronasal organ, is generally well developed in squamates and is crucial for detection of pheromones and prey odors. Despite the numerous studies on embryonic development of the VSE in squamates, especially in snakes, an ultrastructural analysis, as far as we know, has never been performed. Therefore, we investigated the embryology of the VSE of the grass snake (Natrix natrix) using electron microscopy (SEM and TEM) and light microscopy. As was shown for adult snakes, the hypertrophied ophidian VSE may provide great resolution of changes in neuron morphology located at various epithelial levels. The results of this study suggest that different populations of stem/progenitor cells occur at the base of the ophidian VSE during embryonic development. One of them may be radial glia-like cells, described previously in mouse. The various structure and ultrastructure of neurons located at different parts of the VSE provide evidence for neuronal maturation and aging. Based on these results, a few nonmutually exclusive hypotheses explaining the formation of the peculiar columnar organization of the VSE in snakes were proposed.

Sexual segment of the kidney of the lizard, Eutropis carinata: A light microscopic and ultrastructural seasonal study.

We studied seasonal variation of the secretory granules in the epithelial cells of the sexual segment of the kidney (SSK) during the annual sexual cycle in the lizard, Eutropis carinata using light and electron microscopy in correlation with measurements of androgen levels. During the breeding phase, the epithelium of the SSK consists of simple columnar cells with basal nuclei. The cytoplasm contains numerous eosinophilic secretory protein and carbohydrate granules, but lacks glycosaminoglycans. These secretory granules develop during the regenerative phase when the circulating testosterone level increase. During the breeding phase, when the circulating testosterone levels are high, three types of secretory granules can be differentiated in the cytoplasm based on size and opacity; electron translucent type I, electron dense type II, and biphasic type III granules. Type II granules are found at various stages of maturity and degeneration/utilization. All types of secretory granules are released through an apocrine process. Microvilli and tight junctions are prominent at the apical portion of the cell. The cytoplasm contains, Golgi complexes, an abundant network of rough endoplasmic reticulum, numerous tubular mitochondria, condensing, mucus filled and empty vacuoles. Intercellular canaliculi are narrow and indistinct during the regenerative and breeding phases, respectively. During the regressed phase, when the circulating testosterone levels are lowest, the cells are found regressed with wide intercellular canaliculi and devoid of secretory granules. Then the cytoplasm contains a few round mitochondria, Golgi and scanty endoplasmic reticulum.