cDNA characterization of the ribosomal protein L10a gene and its functional analysis in ovarian development of Macrobrachium nipponense

Abstract

Ribosomal proteins (RPs) not only play an important role in protein synthesis but also have extraribosomal functions. In this study, the entire cDNA sequence of the RPL10a gene of Macrobrachium nipponense (MnRPL10a) was cloned by RACE-PCR. The obtained complete cDNA sequence of the MnRPL10a gene was 885 bp encoding 217 amino acids (AAs). The expression pattern of the MnRPL10a gene was analysed by quantitative real-time fluorescence PCR (qPCR), and its mRNA location was determined by fluorescence in situ hybridization (FISH). The results showed that during development, the highest expression of MnRPL10a in the fertilized ovum. Among tissues, it was mostly expressed in the ovary, and its highest expression was found in stage V ovaries. MnRPL10a mRNA was mainly distributed in the cytoplasm of oogonia and oocytes. The role of the MnRPL10a gene in ovarian development was verified by RNA interference (RNAi) and ovarian explant incubation with 5-hydroxytryptamine (5-HT) and dopamine (DA) experiments. The results indicated that the expression of the genes that positively regulate ovarian development, including vitellogenin (Vg), vitellogenin receptor (Vgr), cell cycle protein B (CyclinB) and cell division cyclin 2 (cdc2), decreased, while the expression of the genes that negatively regulate ovarian development, including Wee1 kinase (Wee1) and OTU domain-containing ubiquitin aldehyde-binding protein B (OTUB), increased with the decrease in MnRPL10a expression after RNAi and DA incubation. However, 5-HT incubation showed the opposite results. In summary, the MnRPL10a gene may regulate the development of the ovary in M. nipponense by affecting vitellin synthesis, oocyte meiosis and the oestrogen signalling pathway.

Keywords

• Macrobrachium nipponense
• Ovarian development
• Ribosomal protein L10a gene
• RNA interference
• Ovarian explant incubation