Cellular Evidence of Exosomes in the Reproductive Tract of Chinese Soft-Shelled Turtle Pelodiscus sinensis.

The oviduct is a location of egg production, fertilization, and sperm storage. While its secretions have broadly attributes toward different physiological functions. We examined the ultrastructure of oviduct epithelium and glands in relation to the secretions, particularly with exosomes origin in Chinese soft-shelled turtle Pelodiscus sinensis using immunohistochemistry and transmission electron microscopy. The ciliated epithelial and gland cells were involved in the release of exosomes and secretions into lumen throughout the year. The exosomes were either released directly from epithelium or in relation with multivesicular body (MVB). The average size of the particles varies between 50 and 130 nm. These exosomes were also widely distributed in the epithelial ciliated cells and pericytoplasm of glands lumen. Intracellular MVB was characterized by membrane-bounded exosomes of different sizes. Exosomes were also found in close contact with the cilia and sperm membrane in the lumen, which is suggestive of their fusogenic properties. Immunohistochemistry results showed strong to moderate positive expression of exosomes, in ciliated and gland cells, during January, September, and December, as it is the time of sperm storage in this turtle, whereas they showed moderate to weak level of expression during breeding season (May). This is first study about identification of the exosomes in female turtles. Epithelial and glandular exosomes, intracellular MVB, secretions, and secretory vesicles give this turtle specie a unique secretory morphology and a potential model for investigating the secretory nature of the oviduct.

Waqas MY ，Zhang Q ，Ahmed N ，Yang P ，Xing G ，Akhtar M ，Basit A ，Liu T ，Hong C ，Arshad M ，Rahman HMS ，Chen Q ... -  《-》

Novel cellular evidence of oviduct secretions in the Chinese soft-shelled turtle Pelodiscus sinensis.

The oviduct is the location of fertilization and sperm storage. We examined the ultrastructure of the oviduct epithelium and its glandular secretions in the isthmus, uterus and vagina of Chinese soft-shelled turtle Pelodiscus sinensis using light and transmission electron microscopy. The epithelium in these segments is lined with ciliated, secretory and other cells; the first two cell types span the entire epithelium, with secretory cells being predominant. The ciliated cells are characterized by the presence of a secretory vacuole that releases apocrine secretions into the lumen, whereas the secretory cells contain typical biphasic granules with both dark and light aspects. The third type of cells observed have wider proximal portion, abundant mitochondria, vacuoles, and narrow nuclei. The storage of spermatozoa is restricted to the isthmus, uterus, and vagina. In addition, the gland cells show prominent features, including the presence of granules of different shapes, sizes, and electron densities. The synthesis of these granules is described for the first time in this study. Mitochondria appear to play an important role in the formation of dense granules, the rough endoplasmic reticulum and microfilaments may also play a role in the maturation of these dense granules. After completing the maturation process, these granules are released into the lumen of the gland cells.

Waqas MY ，Lisi H ，Yang P ，Ullah S ，Zhang L ，Zhang Q ，Li Q ，Ahmad N ，Chen W ，Zeshan B ，Chen Q ... -  《-》

Ultrastructure of anterior uterus of the oviduct and the stored sperm in female soft-shelled turtle, Trionyx sinensis.

Ultrastructure of sperm storage in female soft-shelled turtle, Trionyx sinensis was examined under light and electron microscopes. Sperm storage tubules are restricted to the anterior of the uterus. These tubules developed either by folding or fusion of the oviductal mucosal folds and are lined by both ciliated and secretory cells. Ciliated cells are characterized by a few microvilli and prominent cilia in the apical membranes. A prominent feature of the secretory cell is the presence of secretory granules in the supranuclear region. The size, shape, and electron density of these granules vary markedly. The secretory product is released mainly by exocytosis into the oviductal lumen, where it appears as flocculent material. The unique structure in the base of the epithelium, the basal border of the cell -- the basal lamina -- and a blood vessel layer, is presumed to be a important barrier, by which the nourishment exchange and the microenvironment maintenance are ensured. The gland cell is presented with numerous, round, membrane-bound secretory granules of moderate to high electron densities. We divide these granules into three types according to their appearance: (1) membrane bounded granules with high-homogeneous electron density, (2) membrane bounded granules with moderate-homogeneous electron density, (3) membrane bounded, electron dense granules with concentric structures. These granules are presented as different stages of the secretions in the gland cell. The junction complexes are markedly distributed between cells, which are important in keeping stability and the microenvironment maintenance of the sperm storage tubules. Sperm stored in the tubules are heterogeneous in cytology. In addition to the mature sperm in the lumen, sperm with large chromatic granules are found, which are presumed to be immature sperm and are being in the process of nuclear condensation. Several spermatozoa in the tubules are exhibited with definitive indications of degeneration of the nuclei. The nuclear volume increases. The electron density of the central cores in mitochondria declines, combined with the deterioration of concentric membrane structure. Those changes are possibly due to the long time storage of the sperm in sperm storage tubules, and the leakage of reactive oxygen species is suggested to be a major cause. We conclude that the ultrastructure character of sperm storage in the oviduct of Trionyx sinensis is unique, in addition to having a basal function in secretion and the cilia swing, the tubules also provide an available microenvironment for the sperm to long time stored. The degenerative sperm in the tubules might be related to paternity-specific reproductive adaptations, and the sperm competition might occur during long time storage.

Han X ，Zhangli L ，Li M ，Bao H ，Hei N ，Chen Q ... -  《Anatomical Record-Advances in Integrative Anatomy and Evolutionary Biology》

Characterization of the ultrastructure in the uterovaginal junction of the hen.

In poultry, the infundibulum is the place of fertilization, eggshell production, and sperm storage, while its uterovaginal junction (UVJ) is regarded as the most important site, which has abundant sperm storage tubules (SST). We examined the ultrastructure of the epithelium with relation to its unique secretory cytology in the UVJ of hens using transmission electron microscopy. The epithelium of the UVJ is lined with ciliated and secretory cells. Ciliated cells are characterized with light and dense secretory granules in supernuclear cytoplasm. Dense secretory granules in ciliated cells are larger in diameter (one μm), surrounded with a transparent rim and concentric layers, whereas the dense granules in the ciliated cells of SST are smaller (0.52 μm) in size and not surrounded by any transparent rim or layer. Ciliated cells also are involved in the shedding of exosomes and secretory vesicles in the lumen. Secretory exosomes are in close contact with cilia and directly release from the apical border into the lumen. Cell junctions are widely distributed between these cells. The secretory cells are associated with the release of secretions via apocrine blebs from microvilli and secretory vesicles, which protrude out from the surface of the epithelium. The dense secretory granules in these cells are smaller in size (0.6 μm), absent of a transparent rim or layers, and are released into the lumen through secretory vesicles. The intracellular multivesicular body (MVB) also is observed in the supernuclear cytoplasm of secretory cells, which are related to the production of exosomes. In general, the apical protrusion of the epithelium in the form of apocrine secretions, the releasing of exosomes, the identification of intracellular MVB, and the release of dense granules give the epithelium a distinct morphology in the UVJ of the hen oviduct.

Waqas MY ，Yang P ，Ahmed N ，Zhang Q ，Liu T ，Li Q ，Hu L ，Hong C ，Chen Q ... -  《POULTRY SCIENCE》

Identification and characterization of telocytes in the uterus of the oviduct in the Chinese soft-shelled turtle, Pelodiscus sinensis: TEM evidence.

Telocytes (Tcs) are cells with telopodes (Tps), which are very long cellular extensions with alternating thin segments (podomers) and dilated bead-like thick regions known as podoms. Tcs are a distinct category of interstitial cells and have been identified in many mammalian organs including heart, lung and kidney. The present study investigates the existence, ultrastructure, distribution and contacts of Tcs with surrounding cells in the uterus (shell gland) of the oviduct of the Chinese soft-shelled turtle, Pelodiscus sinensis. Samples from the uterine segment of the oviduct were examined by transmission electron microscopy. Tcs were mainly located in the lamina propria beneath the simple columnar epithelium of the uterus and were situated close to nerve endings, capillaries, collagen fibres and secretory glands. The complete morphology of Tcs and Tps was clearly observed and our data confirmed the existence of Tcs in the uterus of the Chinese soft-shelled turtle Pelodiscus sinensis. Our results suggest these cells contribute to the function of the secretory glands and contraction of the uterus.

Ullah S ，Yang P ，Zhang L ，Zhang Q ，Liu Y ，Chen W ，Waqas Y ，Le Y ，Chen B ，Chen Q ... -  《-》