Examination of the effects of impaired spermatogenesis on sterility in triploid oysters (Crassostrea gigas), and implications for commercial aquaculture

Sterility caused by triploidization holds promising commercial implications in aquaculture. In this study, we conducted a thorough investigation into the spermatogenesis of triploid male oysters (Crassostrea gigas) to evaluate their fertility. Histological analysis and immunostaining revealed the presence of numerous immature spermatocytes and a markedly low abundance of mature spermatozoa in triploid males, indicating a robust blockade in spermatogenesis. Ultrastructural analysis and apoptosis analysis demonstrated that triploid males underwent normal spermatogonial mitosis during early spermatogenesis, like diploids. Analysis of phosphorylated histone H2AX (γ-H2AX) displayed heightened γ-H2AX staining in developed triploid male gonads, with a significantly higher number of apoptotic cells confirmed by the TUNEL assay compared to diploid counterparts. These findings suggest a potential arrest at the zygotene stage, preventing the completion of the zygotene-pachytene transition and ultimately leading to cell apoptosis. Flow cytometry results confirmed aneuploidy in spermatozoa from triploid oysters, containing approximately 1.5 times the DNA content of diploid oysters. Notably, the sperm head size and flagella length in triploid oysters surpassed those in their diploid counterparts. Occasional observations of two-tailed spermatozoa in triploid males were noted. This study provides novel insights into the blocked spermatogenesis in triploid oysters, emphasizing the sterility of triploid male oysters and its relevance in aquaculture practices.