Steroidogenic acute regulatory protein transcript abundance in the eel, Anguilla australis: changes during the induced reproductive cycle and effects of follicle-stimulating hormone during previtellogenesis

Abstract

Steroidogenic acute regulatory protein (StAR) mRNA levels in the eel ovary were assayed by quantitative PCR and related to plasma steroid levels throughout oogenesis in order to shed light on the previously considered 'aberrant' prematurational increase in plasma levels of estradiol-17β (E2). Total ovarian StAR transcript abundance mirrored circulating levels of E2, but not of 11-ketotestosterone (11KT). The study was complemented by evaluation of in vitro effects of follicle-stimulating hormone (FSH) on ovarian StAR transcript abundance and on short-term ('acute') radiolabelled pregnenolone-supported steroid metabolism by ovarian fragments to understand how the production of steroids during previtellogenic oocyte growth is regulated. We observed a significant effect of FSH on StAR mRNA levels within 24h of incubation, but these were no longer evident by 4 days of culture. Unexpectedly, FSH had no effect on substrate-supported steroidogenesis, as comparable yields of steroid products were detected using semi-quantitative HPLC and scintillation counting. We conclude that the eel ovarian follicle can respond to FSH from a very early stage of development (early oil droplet stage) by increasing StAR mRNA levels, but that there is no evidence for acute effects of FSH on bioactive steroid production downstream of cytochrome P450 side-chain cleavage. Furthermore, the prematurational increase in StAR mRNA in vivo is in keeping with general teleost models and is likely to be a 'normal' response to reaching advanced stages of development.

Keywords: 11-OHA; 11-ketotestosterone; 11KT; 11β-hydroxyandrostenedione; 17,20β-P4; 17,20β-dihydroxy-4-pregnen-3-one; E2; EV; Eel; Ef1a; FSH; HPLC; HSD; Induced maturation; LH; MN; MV; Ovary; P450scc; PV; Previtellogenesis; SPH; StAR; Steroidogenesis; T; cytochrome P450 side-chain cleavage; early vitellogenic; elongation factor-1α; estradiol-17β; follicle-stimulating hormone; high pressure liquid chromatography; hydroxysteroid dehydrogenase; luteinizing hormone; midvitellogenic; migratory nucleus; previtellogenic; qPCR; quantitative polymerase chain reaction; salmon pituitary homogenate; steroidogenic acute regulatory protein; testosterone.