title: Global analysis of differential gene expression related to long-term sperm storage in oviduct of Chinese Soft-Shelled Turtle Pelodiscus sinensis.
abstract: Important evolutionary and ecological consequences arise from the ability of female turtles to store viable spermatozoa for an extended period. Although previous morphological studies have observed the localization of spermatozoa in Pelodiscus sinensis oviduct, no systematic study on the identification of genes that are involved in long-term sperm storage has been performed. In this study, the oviduct of P. sinensis at different phases (reproductive and hibernation seasons) was prepared for RNA-Seq and gene expression profiling. In total, 2,662 differentially expressed genes (DEGs) including 1,224 up- and 1,438 down-regulated genes were identified from two cDNA libraries. Functional enrichment analysis indicated that many genes were predominantly involved in the immune response, apoptosis pathway and regulation of autophagy. RT-qPCR, ELISA, western blot and IHC analyses showed that the expression profiles of mRNA and protein in selected DEGs were in consistent with results from RNA-Seq analysis. Remarkably, TUNEL analysis revealed the reduced number of apoptotic cells during sperm storage. IHC and TEM analyses found that autophagy occurred in the oviduct epithelial cells, where the spermatozoa were closely attached. The outcomes of this study provide fundamental insights into the complex sperm storage regulatory process and facilitate elucidating the mechanism of sperm storage in P. sinensis.
authors.affiliation: Laboratory of Animal Cell Biology and Embryology, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu Province, 210095, China.
authors.forename: Tengfei
authors.initials: T
authors.lastname: Liu
authors.affiliation: Laboratory of Animal Cell Biology and Embryology, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu Province, 210095, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Yang
authors.affiliation: Laboratory of Animal Cell Biology and Embryology, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu Province, 210095, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Chen
authors.affiliation: Laboratory of Animal Cell Biology and Embryology, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu Province, 210095, China.
authors.forename: Yufei
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Laboratory of Animal Cell Biology and Embryology, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu Province, 210095, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Laboratory of Animal Cell Biology and Embryology, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu Province, 210095, China.
authors.forename: Yasir
authors.initials: Y
authors.lastname: Waqas
authors.affiliation: Laboratory of Animal Cell Biology and Embryology, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu Province, 210095, China.
authors.forename: Nisar
authors.initials: N
authors.lastname: Ahmed
authors.affiliation: Laboratory of Animal Cell Biology and Embryology, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu Province, 210095, China.
authors.forename: Xiaoya
authors.initials: X
authors.lastname: Chu
authors.affiliation: Laboratory of Animal Cell Biology and Embryology, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu Province, 210095, China.
authors.forename: Qiusheng
authors.initials: Q
authors.lastname: Chen
mesh_terms: D000818:Animals; D004822:Epididymis / Q000254:growth & development / Q000378:metabolism; D005187:Fallopian Tubes / Q000254:growth & development; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000235:genetics; D006605:Hibernation / Q000235:genetics; D008297:Male; D010057:Oviducts / Q000254:growth & development / Q000378:metabolism; D012098:Reproduction / Q000235:genetics*; D013094:Spermatozoa / Q000254:growth & development* / Q000378:metabolism; D014426:Turtles / Q000235:genetics* / Q000254:growth & development