Effects of pre-overwintering conditions on eupyrene and apyrene spermatogenesis after overwintering in Polygonia c-aureum (Lepidoptera: Nymphalidae)

Sperm polymorphism is widely known in invertebrates. In insects, Lepidoptera has two types of sperm: nucleated eupyrene (fertile) sperm and anucleated apyrene (unfertile) sperm. These sperm types are produced during post-embryogenesis, and eupyrene spermatogenesis precedes apyrene spermatogenesis. During overwintering, spermatogenesis stops and a portion of undifferentiated-stage spermatocytes degenerate. After overwintering, spermatogenesis restarts with unaffected spermatogonia. However, how new spermatozoa arise in the adult testes after overwintering is not known in Lepidoptera. In this study, we investigated the spermatogenesis events in the nymphalid butterfly Polygonia c-aureum after overwintering under various environmental conditions. Our results indicate that both eupyrene and apyrene spermatogenesis restart at any stopping stage and sperm of these types are regenerated in no particular order after adult insect overwintering. This suggests that the spermatogenesis occurring after overwintering proceeds without embryogenetic restrictions related to the developmental sequence.