Analysis and gonadal localization of Speedy A mRNA transcript, a novel gene associated with early germline cells in the scallop, Argopecten purpuratus

Abstract

The Speedy A (spdya) gene is a member of the Speedy/RINGO family, encoding a spdya protein associated with cellular cycle and meiosis in vertebrates. Results from genetic analyses indicated spdya conditional knockout mice are sterile, suggesting that this protein has essential functions in mammalian reproduction. There, however, are no published reports on the localization of spdya mRNA in the germline or in somatic cell lineages within the gonads from mollusks or other invertebrate species. Using a previously obtained transcriptome assembly from the scallop Argopecten purpuratus, an economically important hermaphroditic scallop species from Chile and Peru, there was identification of a complete coding sequence of the spdya mRNA. Phylogenetically spdya protein has sequence conservation homology with other scallops and mollusks. The relative mRNA transcript abundances at different gametogenic stages was assessed using quantitative PCR procedures. Results indicated there was an increase of spdya mRNA transcript abundance in testicular region samples at the late active stage, followed by a decrease in testis of reproductively mature individuals. To gain insight into the cellular localization of ap-spdya transcript within the gonads, specific RNA probes were synthesized for in situ hybridization analyses of gonad histological sections. Results indicated spdya mRNA is located exclusively in early germline (previtellogenic oocytes and spermatogonia) and somatic proliferative tissues of A. purpuratus ovarian and testicular regions. Overall, these results indicate there are putative functions of spdya in the early oogenesis and spermatogenesis of A. purpuratus and will contribute to furthering the understanding of gametogenesis in this species.