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Spermatogenesis in reptiles is a seasonally dependent physiological process that is not temporally associated with male mating behavior. Characteristics of seasonal spermatogenesis in reptiles, however, remain largely unknown. In this review, there is a coverage of the characteristics of soft-shelled turtle, Pelodiscus sinensis, during seasonal spermatogenesis that provides insights into spermatogenesis of testudines. The seminiferous epithelium of P. sinensis are undergoing spermatogenesis during the summer and fall, but are quiescent throughout the rest of the year; germ cells progress through spermatogenic stages in a temporal rather than a spatial pattern. While apoptotic germ cells mainly appear in the non-spermatogenic phase, these are seldom present during active spermatogenesis. It is inferred that apoptosis may be one of the reasons for germ cell loss during the resting phase of spermatogenesis. During the period when spermatogenesis is occurring, Sertoli cells become very narrow and are in contact with several round/elongated spermatids. Many residual spermatozoa can be internalized and degraded within Sertoli cells by entosis during the non-spermatogenic phase, which precedes the next reproductive cycle in P. sinensis. In the late spermatogenic phase, round-shaped mitochondria of spermatids become elongated and swollen, subsequently forming a crescent-like shape and develop into "onion-like" shaped mitochondria. As spermiogenesis progresses, the endoplasmic reticulum of spermatids is transferred into a specialized structure called the "Chrysanthemum flower center", which may be a source of autophagosomal membranes. The information provided in this review will help improve understanding of characteristics of seasonal spermatogenesis, which will hopefully promote interest in the study of reptilian species.

Apoptosis is a physiological phenomenon that has been recognized as a cause of sperm death during cryopreservation in endothermic mammals. There is, however, no data on its role in sperm death during cooled storage in ectothermic animals. In this study, spermatozoa from the epididymis of soft-shelled turtle were investigated to identify the mechanism of spermatozoa apoptotic-like changes during storage at 4 °C. In this study, there was survival of spermatozoa for more than 40 Days when stored at 4 °C. During cooled storage, sperm kinematics was evaluated using CASA system. Values for all sperm motility variables decreased during the period of storage; while for velocity curvilinear (VCL) there was a further decrease after 20 Days of storage. Results from flow cytometry analysis indicated that there was a significant increase in the percentage of apoptotic spermatozoa, but there was no change in the percentage of necrosis. Furthermore, the concentration of cellular ROS increased after 20 Days of storage at 4 °C. The results using JC-1 staining indicated there was a decrease in MMP of spermatozoa as the duration of storage at 4 °C increased. Nuclear fragmentation of spermatozoa was observed using TEM on Day 30 of storage. There were large amounts of pro-apoptotic cytochrome c (Cytc) and cleaved caspase-9/3 proteins detected using western blot analysis after 30 days of spermatozoa storage at 4 °C. These findings indicate ROS generation induces mitochondria damage after 20 days of storage at 4 °C, which can induce spermatozoa apoptotic-like changes during storage of soft-shelled turtle spermatozoa.