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Gem-associated protein 2-like isoform X1 ( GEM ) was previously predicted to be involved in the sexual development of male Macrobrachium nipponense . In this study, we analyze the GEM functions in depth using quantitative polymerase chain reaction (qPCR), in situ hybridization, and RNA interference (RNAi). The full-length Mn-GEM cDNA sequence was 1018 base pairs (bp) long with an open reading frame of 777 bp encoding 258 amino acids. qPCR analysis of Mn-GEM in different tissues and developmental stages showed that Mn-GEM was highly expressed in the gonad and from post-larval developmental stage day 5 (PL5) to PL15, which indicated that GEM has potential roles in gonad differentiation and development in M . nipponense . In situ hybridization and qPCR analysis of various stages of the reproductive cycle of the testis and ovary indicated that GEM may promote spermatid development and gametogenesis in M . nipponense . After injecting with double-stranded RNA (dsRNA) of Mn-GEM , mRNA expression of Mn-insulin-like androgenic gland hormone ( Mn-IAG ) and the content of testosterone increased with the decrease of Mn-GEM expression, indicating that GEM has negative effects on the male sexual differentiation and development in M . nipponense . Results of this study highlight the functions of GEM in M . nipponense , which can be applied to future studies of male sexual development in M . nipponense and other crustacean species.

international journal of molecular sciences

Potential Functions of Gem-Associated Protein 2-Like Isoform X1 in the Oriental River Prawn Macrobrachium nipponense: Cloning, qPCR, In Situ Hybridization, and RNAi Analysis