Functional characterization of a cDNA encoding Piwi protein in Penaeus monodon and its potential roles in controlling transposon expression and spermatogenesis

摘要

Piwi proteins comprise a subfamily of Argonaute that plays a major role in germline development by association with a distinct class of small RNAs called Piwi interacting RNA (piRNA). Although the functions of Piwi in the development of germline cells as well as transposon regulation were reported in a number of mammalians and insects, developmental expression and function of Piwi subfamily in crustaceans is poorly known. This study is aimed at cloning and characterization of a Piwi cDNA in the black tiger shrimp, Penaeus monodon. The cDNA encoding a Piwi protein of P. monodon (PmPiwi1) was obtained by rapid amplification of cDNA ends (RACE). The PmPiwi1 coding cDNA contains 2811 nt encoding a putative protein of 936 amino acids, and was specifically expressed in testis and ovary, suggesting its possible function in gametogenesis. RNAi experiment showed that suppression of PmPiwi1 expression led to a significant up-regulation of retrotransposon gypsy2 and DNA element transposon mariner in shrimp testis. Investigation of the function of PnsPiwi1 in spermatogenesis by sperm count showed significantly lower number of sperms in the spermatophore sac of PmPiwi1-knockdown shrimp compared with that in the control shrimp. Our study thus reported for the first time the cDNA encoding a Piwi protein in the shrimp P. monodon. Its roles in controlling transposons and spermatogenesis as implied by the results in this study will be important for understanding sperm development and could be useful for the improvement of reproduction in male shrimp in the future.