Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
2072575 | Animal Reproduction Science | 2016 | 10 Pages |
•A cDNA (Sp-RPCH), cloned from the mud crab Scylla paramamosain, encodes a PRCH precursor and is expressed in nervous tissues.•Sp-RPCH expression levels in nervous tissues fluctuated along a cycle of ovarian maturation.•Synthetic RPCH evoked a Ca2+ current in dissociated brain neurons.•Synthetic RPCH in vitro significantly increased mean oocyte diameter of ovarian tissues co-cultured with nervous tissues.•Synthetic RPCH in vivo significantly increased gonad-somatic index, hepatopancreas-somatic index, and vitellogenin expression in ovary and hepatopancreas.
In this study a full-length cDNA (Sp-RPCH) was cloned from the eyestalk ganglia of the mud crab Scylla paramamosain. Sp-RPCH is 660 base pairs in length and its open reading frame encodes a precursor that is predicted to be processed into a 25-residue signal peptide, a mature red pigment concentrating hormone (RPCH, an octapeptide), and a 75-residue precursor-related peptide. Phylogenetic analysis indicates that it clusters with other crustacean RPCHs and belongs to the adipokinetic hormone/RPCH peptide superfamily. Sp-RPCH gene expression was detected, using an end-point polymerase chain reaction (PCR), not only in the eyestalk ganglia but also in the brain and thoracic ganglia. Quantified using a real-time PCR, Sp-RPCH gene expression levels in the three tissues fluctuated along a cycle of ovarian maturation, with the levels progressively increased from stages I to IV, after which the expression levels decreased (although they remained significantly higher than stage I levels) when the ovary reached the mature stage (stage V). It was demonstrated using a patch clamp analysis that synthetic RPCH was able to evoke a Ca2+ current in dissociated brain neurons and synthetic RPCH significantly increased the mean oocyte diameter of the ovarian tissues co-cultured with the eyestalk ganglia, brain, or thoracic ganglia; the stimulatory effect of RPCH was absent when the nervous tissues were not included in the ovarian incubation. Animals administrated with RPCH had significantly higher levels of gonad-somatic index, hepatopancreas-somatic index, and vitellogenin gene expression, when compared to control animals receiving a saline injection. The combined results clearly show that RPCH is involved in ovarian maturation in the mud crab; the stimulatory effects of RPCH are likely mediated by its actions on the release from the nervous tissues of factor(s) that directly regulate vitellogenesis in the ovary and hepatopancreas.