Seasonal spermatogenesis in the red-eared slider (Trachemys scripta elegans): The roles of GnRH, actin cytoskeleton, and MAPK

Reproduction is the key to the ecological invasion of alien species. As an invasive species, the characteristic and regularity of red-eared slider (Trachemys scripta elegans) spermatogenesis is an index for evaluating reproduction and ecological adaptation. Here, we investigated the characteristics of spermatogenesis i.e., the gonadosomatic index (GSI), plasma reproductive hormone levels, and the histological structure of testes by HE and TUNEL staining, and then RNA-Seq in T. s. elegans. The histomorphological evidence confirmed that seasonal spermatogenesis in T. s. elegans has four successive phases: quiescence (December-May of the following year), early-stage (June-July), mid-stage (August-September), and late-stage (October-November). In contrast to 17β-estradiol, testosterone levels were higher during quiescence (breeding season) compared to mid-stage (non-breeding season). Based on RNA-seq transcriptional analysis, gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were used to analyze the testis in the quiescent and mid-stage. Our study found that circannual spermatogenesis is regulated by interactive networks including gonadotropin-releasing hormone (GnRH) secretion, regulation of actin cytoskeleton, and MAPK signaling pathways. Moreover, the number of genes associated with proliferation and differentiation (srf, nr4a1), cell cycle (ppard, ccnb2), and apoptosis (xiap) were up-regulated in the mid-stage. With the maximum energy saving, this seasonal pattern of T. s. elegans determines optimal reproductive success and thus adapts better to the environment. These results provide the basis for the invasion mechanism of T. s. elegans and lay the foundation for deeper insight into the molecular mechanism of seasonal spermatogenesis in reptiles.