Seasonal effects on apoptosis and proliferation of germ cells in the testes of the Chinese soft-shelled turtle, Pelodiscus sinensis

Abstract

To elucidate the processes involved in the spatial and temporal maturation of spermatogenic cells in the testes of the soft-shelled turtle, Pelodiscus sinensis, we used a histological morphology method, TdT-mediated dUTP nick end-labeling (TUNEL) assay, the proliferating-cell nuclear antigen (PCNA), and electron microscopy. Seminiferous tubules from 100 turtles, normal for size of testes and semen quality, were collected during 10 months of a complete annual cycle (10 turtles/month). The seminiferous epithelium was spermatogenically active through the summer and fall, but quiescent throughout the rest of the year; germ cells progressed through spermatogenesis in a temporal rather than a spatial pattern, resulting in a single spermatogenic event that climaxed with one massive sperm release in November. The TUNEL method detected few apoptotic cells in spermatogenic testis, with much larger numbers during the spermatogenically quiescent phase. Spermatocytes were the most common germ cell types labeled by the TUNEL assay (a few spermatogonia were also labeled). Apoptotic spermatocytes had membrane blebbing and chromatin condensation during the resting phase, but not during active spermatogenesis. We inferred that accelerated apoptosis of spermatogonia and spermatocytes partly accounted for germ cell loss during the nonspermatogenic phase. The PCNA was expressed in nuclei of spermatogonia and primary spermatocytes during the spermatogenically active phase. During the regressive phase, PCNA-positive cells also included spermatogonia and spermatocytes, but the number of positive spermatocytes was less than that during the spermatogenically active phase. We concluded that seasonal variations in spermatogenesis in the soft-shelled turtle were both stage- and process-specific.