Molecular cloning, characterization and expression profile of kisspeptin1 and kisspeptin1 receptor at brainpituitary-gonad (BPG) axis of golden mahseer, Tor putitora (Hamilton, 1822) during gonadal development

Abstract

Complete cDNA sequences of kiss1 (gmkiss1) and its receptor kiss1r (gmkiss1r) were cloned and characterized from brain tissue of adult golden mahseer (Tor putitora). Thereafter, quantification of gmkiss1 and gmkiss1r mRNA expression in brain-pituitary-gonad (BPG) axis of male and female golden mahseer was carried out using quantitative real-time (qRT)-PCR assay during an annual reproductive cycle, at different gonadal development stages. The gmkiss1 cDNA was 508 bp, with 330 bp open reading frame (ORF), encoding a precursor protein of 109 amino acids, whereas gmkiss1r cDNA was 1383 bp with an ORF of 1004 bp, which encodes a 334 amino acid protein residue. The qRT-PCR study shows that gmkiss1 and gmkiss1r are expressed in brain, pituitary and gonads of both the sexes of golden mahseer. An apparent sexual dimorphism in transcript level of gmkiss1 and gmkiss1r in brain and gonads was evident during the reproductive cycle. Overall, in brain, testis and ovary, the gmkiss1 and gmkiss1r mRNA expression level was comparatively higher during the initial stages of gonadal development, than that of spermiation or ovulation stage. In pituitary of both the sexes, throughout the gonadal development, consistently low transcript level of gmkiss1 and gmkiss1r was observed. The gmkiss1 mRNA expression level in brain and ovary of female golden mahseer was several folds higher than the brain and testis of male fish. In conclusion, we confirm the presence of kiss1 and its receptor in golden mahseer, and results of our study strongly suggested the involvement of kisspeptin1 system in gonadal development and annual reproductive cycle of this species.