Development of a flatfish-specific enzyme-linked immunosorbent assay for Fsh using a recombinant chimeric gonadotropin

摘要: In flatfishes with asynchronous and semicystic spermatogenesis, such as the Senegalese sole (Solea senegalensis), specific roles of pituitary gonadotropins during germ cell development, particularly follicle-stimulating hormone (Fsh), are still largely unknown in part due to lack homologous immunoassays for this hormone. study, an enzyme-linked immunosorbent assay (ELISA) Fsh was developed by generating a rabbit antiserum against recombinant chimeric single-chain molecule (rFsh-C) produced yeast Pichia pastoris. The rFsh-C N- C-termini were formed mature β subunit (Fshβ) chicken glycoprotein common α (CGA), respectively. Depletion remove anti-CGA antibodies further enriched Fshβ-specific antibodies, which used develop ELISA using standard curve. sensitivity 10 50 pg/ml measurement plasma pituitary, respectively, cross-reactivity luteinizing 1%. curve paralleled those serially diluted extracts other flatfishes, Atlantic halibut, turbot. males, highest levels found early spermatogenesis but declined enhanced spermiation, teleosts cystic spermatogenesis. pubertal however, circulating high adult spermiating fish, interestingly receptor developing testis containing only spermatogonia expressed Leydig cells not primordial Sertoli cells. These results indicate that can be generate Fshβ highly sensitive measurements diverse flatfishes.