AUTHOR=Zhang Wenyi , Xiong Yiwei , Wang Pengchao , Chen Tianyong , Jiang Sufei , Qiao Hui , Gong Yongsheng , Wu Yan , Jin Shubo , Fu Hongtuo 

TITLE=RNA interference analysis of potential functions of cyclin A in the reproductive development of male oriental river prawns (Macrobrachium nipponense)

JOURNAL=Frontiers in Genetics

VOLUME=13

YEAR=2022

URL=https://www.frontiersin.org/journals/genetics/articles/10.3389/fgene.2022.1053826

DOI=10.3389/fgene.2022.1053826

ISSN=1664-8021

ABSTRACT=<p>Cyclin A (<italic>CycA</italic>) plays essential roles in regulating multiple steps of the cell cycle, and it affects gonad development in mammals and invertebrates. Previous RNA interference (RNAi) analysis revealed that knocking-down the expression of <italic>CycA</italic> in female oriental river prawns (<italic>Macrobrachium nipponense</italic>) inhibited ovarian development. <italic>CycA</italic> was also predicted to have regulatory roles in reproductive development of male <italic>M. nipponense</italic> based on significant changes of <italic>Mn-CycA</italic> expression after eyestalk ablation. The goal of this study was to investigate the potential functions of <italic>CycA</italic> in the reproductive development of male <italic>M. nipponense</italic> using RNAi and histological observations. Quantitative real-time PCR analysis revealed that both single-side and double-side eyestalk ablation stimulated the expressions of <italic>Mn-CycA</italic>, and the expression was higher in prawns with double-side eyestalk ablation (<italic>p</italic> &lt; 0.05). <italic>Mn-CycA</italic> expression was significantly higher in the testis and androgenic gland during the reproductive season than during the non-reproductive season (<italic>p</italic> &lt; 0.05). In the RNAi analysis, <italic>Mn-CycA</italic> expression significantly decreased after prawns were injected with <italic>dsCycA</italic>, and the expression of <italic>insulin-like androgenic gland hormone</italic> (<italic>Mn-IAG</italic>) also decreased as <italic>Mn-CycA</italic> expression decreased. This result indicated that <italic>CycA</italic> positively regulated the expression of <italic>IAG</italic> in <italic>M. nipponense</italic>. Histological observations revealed that the number of sperm decreased dramatically to &lt;5% of the total cells in the testis of the <italic>dsCycA</italic>-treated group compared to that of control group on day 14, indicating that knockdown of <italic>Mn-CycA</italic> expression inhibited testis development by affecting the expression of <italic>Mn-IAG</italic> in <italic>M. nipponense</italic>. These results highlighted the functions of <italic>CycA</italic> in male reproductive development of <italic>M. nipponense</italic>, which can be applied to future studies of male reproduction in other crustacean species.</p>