Abstract
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.
Highlights
The oriental river prawn Macrobrachium nipponense (Crustacea; Decapoda; Palaemonidae) is widely distributed in Asian countries [1,2,3,4,5]
We used in situ hybridization to identify the locations of Mn-Gem-associated protein 2-like isoform X1 (GEM) mRNA in the androgenic gland, hepatopancreas, testes, and ovaries during different stages of the reproductive cycle
DiTshcuesasniodnrogenic gland of crustaceans is important in male sexual differentiation and development, and GTEhMe aindtrhoigsegnliacndglwanads porfedcircutesdtacteoanbse ains iimmppoorrttant gienneminalemasleexusaexl udailffdeerevnetloiaptimonentanind Mde.vneiplopponmeennset,[1a6n]d
Summary
The oriental river prawn Macrobrachium nipponense (Crustacea; Decapoda; Palaemonidae) is widely distributed in Asian countries [1,2,3,4,5]. Production of all male progeny on a commercial scale is the long-term goal in M. nipponense aquaculture. The androgenic gland in crustaceans produces hormones and drives male sexual differentiation in crustacean species. It plays essential roles in the establishment of male sexual characteristics and the development of the testes [7]. The genes in the androgenic gland may be the key to understanding the mechanism of male differentiation and development of M. nipponense. T GEM is predicted to play essential roles in male sex differentiation and Thadenedvgeodloaepvl emolofepnthmt iiensnMsttn.uendeipdyptoiosnbetneosieda. Rdiefsfeurletnstotfistshuiess satnuddydewveillol pbme eanptapllisctaagbeles atondfuintutrheestudies of maleresperxoudaulctdiveevceyloclpemofeonvtariniesMan. dnitpesptoens.eTnhsee afunndctoiotnhseorfcGrEuMstawceeraenfusrptheecriedse.termined using in situ hybridization and RNA interference (RNAi). XP_027229945.1 XP_022103928.1 XP_018087535.1 XP_022804116.1 XP_004399834.1 XP_023219163.1 XP_013071285.1 XP_005105416.1 XP_021367489.1 NP_001096228.1 NP_001087945.2
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