Identification and Characterization of Germ Cell Genes <i>Vasa</i> and <i>Nanos-2</i> in the Ovary and Testis of White Crappie (<i>Pomoxis annularis</i>) and the Ovary of Black Crappie (<i>P. nigromaculatus</i>)

Abstract

The vasa gene, encoding an ATP-dependent RNA helicase, and the nanos-2 gene, an RNA-binding protein, are essential for germ cell origination, migration, maintenance, and development in vertebrates and invertebrates. The expression levels of the vasa and nanos-2 genes have not yet been investigated or reported in crappie species. These two genes were partially sequenced and characterized, and their expression patterns were analyzed using reverse-transcription quantitative polymerase chain reaction (RT-qPCR) according to age and sex. The vasa sequences of white crappie (WC) females and males showed significant similarity with the vasa homologs of largemouth bass (Micropterus salmoides; 93.1–93.98%) and smallmouth bass (M. dolomieu; 91.95–92.77%), indicating its conserved nature within the Family Centrarchidae. The vasa sequence of black crappie (BC) females showed significant similarity with the vasa homologs of white crappie (91.67%), largemouth bass (96.10%), smallmouth bass (96.10%), spotted scat (Scatophagus argus; 97.37%), mandarin fish (Siniperca chutasi; 96.15%), Japanese sea bass (Lateolabrax japonicus; 94.87%), lumpfish (Cyclopterus lumpus; 91.95%), southern bluefin tuna (Thunnus maccoyii; 94.74%), large yellow croaker (Larimichthys crocea; 92.21%), and Nile tilapia (Oreochromis niloticus; 92.21%). The nanos-2 sequences of WC females, WC males, and BC females showed significant similarity with the nanos-2 of largemouth bass (92.92–96.36%), smallmouth bass (92.92–96.36%), and mandarin fish (92.66–94.34%). The expression of vasa in BC females was significantly higher at age-2 than at age-1, while WC males and females presented no significant age-related differences. Neither species had a significant difference in nanos-2 gene expression with age. The expression levels of vasa and nanos-2 were significantly higher in WC males than females.

Keywords

• <i>vasa</i>
• <i>nanos-2</i>
• gene expression
• crappie species
• germ cell transplantation
• fish reproduction