Common carp pentraxin gene: Evidence for its role in ovarian differentiation and growth

Abstract

Pentraxins (PTX), belong to an evolutionarily conserved family, containing a PTX protein domain, having role in acute immunological responses and fertility in higher vertebrates. However, information regarding the action of ptx on reproduction is extremely limited in fish. To study this, ptx cDNA was cloned for downstream analysis. Tissue distribution and ontogeny expression analysis indicated the prevalence of ptx in ovary. Varied phase-wise expression during carp ovarian cycle and elevated ptx expression after human chorionic gonadotropin induction, in vitro and in vivo, indicated probable regulation of gonadotropin. In situ hybridization and immunohistochemistry revealed the presence of ptx transcript and protein in the follicular layer of stage-III/IV oocytes indicating a role in ovarian growth. To assess the functional significance of ptx, transient silencing was performed using follicular primary cell culture, in vitro and in common carp, in vivo, through ovary-targeted injection of PEI-siRNA. Transient silencing of ptx-siRNA reduced the expression of various genes/factors related to oogenesis such as transcription factors, several steroidogenic enzymes, and esrs genes. These alterations in expression suggested a plausible role for ptx in ovarian steroidogenesis either, directly or indirectly, which is evident from the changes in the serum estradiol-17β (E2) and 17α,20β-dihydroxyprogesterone levels. Furthermore, downregulation of aromatase activity was also noticed after transient silencing. Increased ptx expression after E2 induced sex reversal to juvenile carp showed the correlative role of ptx during ovarian differentiation and development. Taken together, these findings suggest that ptx exerts an important role during ovarian growth, maturation and/or recrudescence of common carp.