B-cell Lymphoma-2 And Cysteinyl Aspartate Specific Proteinase-3 Localization In The Female Reproductive Tract Of The Chinese Soft-shelled Turtle,Pelpdiscus Sinensis,and Its Relationship With Sperm Storage

The Chinese soft-shelled turtle,Pelodiscus sinensis,serves as one of reptiles,which has a special status in phylogenetic tree.It is also one of major aquatic animals in China and is rich in nutrition.From the economical and pharmacological point of view,it arouses more and more farmers' interests in breeding.Chinese soft-shelled turtle displays a clear dissociation between the male and female gametogenic cycles.This makes prolonged sperm storage be an essential adaptation in reptilian reproductive cycles.In this paper,we choose the Chinese soft-shelled turtle as a model animal and make clear the localization of Bcl-2 and Caspase-3 and where and when sperm storage took place in the oviduct.Aim to provide theoretical basis on the molecular mechanism of sperm storage in the oviduct of Chinese soft-shelled turtle,we explore the relationship of Bcl-2 and Caspase-3 with sperm storage.Experiment I Sperm storage in the female oviduct of the Chinese soft-shelled turtle,Pelodiscus sinensis Hematoxylin-eosin staining was used to stain the oviduct of the female Chinese soft-shelled turtle,Pelodiscus sinensis,to clear where and when sperm storage took place in the oviduct.Ultrastructural detail of the epithelium in the oviduct was observed by the transmission electron microscopy.Hematoxylin-eosin staining showed that female P.sinensis stored sperm in the vagina,uterus and isthmus from November to April of the following year.Sperm came into contact with the epithelial cilia,and some sperm even bored into gland ducts,which is where sperm storage occurred.The result indicated that the epithelium and the secretory activity of gland were related to sperm storage,and it suggested that the secretion of gland and epithelium might form a microenvironment with cilia and it could maintain and promote the sperm storage.Experiment II B-cell lymphoma-a localization in the female reproductive tract of the Chinese soft-shelled turtle,Pelodiscus sinensis,and its relationship with sperm storage The aim of the present study was to investigate the localization of B-cell lymphoma-2(Bcl-2)in the oviduct of the Chinese soft-shelled turtle,Pelodiscus sinensis,during the reproduction cycle to analysis the relationship between Bcl-2 and sperm storage.Oviduct showed positive immunostaining epithelial ciliated cells,gland ducts,and gland cells.Bcl-2 expression in the oviduct was associated with sperm storage occurrence.This indicates that the survival factor Bcl-2 may play a role in P.sinensis sperm storage.Experiment III Cysteinyl aspartate specific proteinase-3 localization in the female reproductive tract of the Chinese soft-shelled turtle,Pelodiscus sinensis,and its relationship with sperm storage The aim of the present study was to investigate localization of cysteinyl aspartate specific proteinase-3(Caspase-3)in the oviduct of the Chinese soft-shelled turtle,Pelodiscus sinensis,during the reproduction cycle to analysis the relationship between Caspase-3 and sperm storage.The localization of Caspase-3 was confirmed by immunohistochemistry.Positive immunostaining was observed in the environment around capillary vessel and cytoplasm of some gland cells during sperm storage and positive immunostaining was observed in the top of epithelial ciliated cells,the nucleus of the ciliated cells,the nucleus of gland cells and environment around capillary vessel when sperm are not stored.Caspase-3 expression in the oviduct was associated with sperm storage occurrence.This indicates that the survival factor Caspase-3 may play a role in P.sinensis sperm storage.