Identification of potentially novel functions of DNA polymerase zeta catalytic subunit in oriental river prawn, Macrobrachium nipoponense: cloning, qPCR, in situ hybridization and RNAi analysis
- 12 Downloads
The goal of this study was to analyze the functions of DNA polymerase zeta catalytic subunit (Rev3) in the oriental river prawn Macrobrachium nipponense (Mn-Rev3) with a focus on its potential roles in sex differentiation and development. The full length of Mn-Rev3 cDNA sequence was 6832 base pairs (bp) with an open reading frame of 6102 bp encoding 2033 amino acids. Mn-Rev3 showed the closest evolutionary relationship with Penaeus vannamei. The highest expression level of Mn-Rev3 occurred in the hepatopancreas and strong signals were observed in hepatopancreas cells, suggesting that Mn-Rev3 played a role in the immune system. Expression levels of Mn-Rev3 also were relatively high in the androgenic gland and testis, suggesting its potential roles in male sexual differentiation and development. During development, expression of Mn-Rev3 was highest on larval day 15 and relatively high from post-larval day 1 (PL1) to PL15, indicating that it played essential roles in promoting metamorphosis and gonad differentiation and development in M. nipponense. Strong Mn-Rev3 signals were detected in spermatids, spermatocytes, and sperm in the testes, and Mn-Rev3 expression was higher in the testes during the reproductive season than in the non-reproductive season. This result indicated that Rev3 promoted whole testis development, and especially sperm development, in M. nipponense. The expression level of Mn-Rev3 was high from ovary V to ovary II stages, indicating that Rev3 may be involved in yolk deposition. The expression level of Mn-insulin-like androgenic gland hormone (Mn-IAG) and the content of testosterone showed the same expression pattern as that of Mn-Rev3 after injection of double-stranded RNA of Mn-Rev3, which indicated that Rev3 had positive effects on male sexual differentiation and development in M. nipponense. The results of this study advance our understanding of male sexual development in M. nipponense and provide the basis for further studies of male sexual differentiation and development in crustaceans.
KeywordsMacrobrachium nipponense Rev3 Sexual differentiation and development RNAi Testosterone
This research was supported by grants from Central Public-interest Scientific Institution Basal Research Fund CAFS (2019JBFM02, 2019JBFM04); the National Key R&D Program of China (2018YFD0900201, 2018YFD0901303); Central Public-interest Scientific Institution Basal Research Fund CAFS (2019JBFM04); Jiangsu Agricultural Industry Technology System (JFRS-02); the National Natural Science Foundation of China (31572617); the China Agriculture Research System-48 (CARS-48); the New cultivar breeding Major Project of Jiangsu province (PZCZ201745).
SJ, HF, YH, and HQ: Conceived and designed the experiments. SJ, YH, YG, and WZ: Performed the experiments. The specimens were maintained by SJ, and YX. SJ: Analyzed the data. YG and YW: Contributed reagents/materials/analysis tools.
Compliance with ethical standards
Conflict of interest
We declare that we do not have any commercial or associative interest that represents a conflict of interest in connection with the work submitted.
All experiments involving M. nipponense in this study have been approved by Institutional Animal Care and Use Ethics Committee of the Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences (Wuxi, China).
- Bureau of Fisheries, Ministry of Agriculture, P.R.C (2016) Fisheries economic statistics. In: Zhao XW (ed) China fishery yearbook. China Agricultural Press, BeijingGoogle Scholar
- Cai Y, Shokita S (2006) Report on a collection of freshwater shrimps (Crustacea: Decapoda: Caridea) from the Philippines, with descriptions of four new species. Raffles Bull Zool 54:245–270Google Scholar
- Cao LQ, Tao YG, Tan HZ (1994) Studies on unisexualization in Red Tipapia. J Fish China. 18(1):75–80Google Scholar
- Jiang FW, Fu HT, Qiao H, Zhang WY, Jiang SF, Xiong YW, Sun SM, Gong YS, Jin SB (2014) The RNA interference regularity of transformer-2 gene of oriental river prawn Macrobrachium nipponense. Chinese Agricult Sci Bull. 30(32):32–37Google Scholar
- Jin SB, Zhang Y, Guan HH, Fu HT, Jiang SF, Xiong YW, Qiao H, Zhang WY, Gong YS, Wu Y (2016) Histological observation of gonadal development during post-larva in oriental river prawn, Macrobrachium nipponense. Chin J Fish 29(4):11–16Google Scholar
- Jin SB, Fu HT, Sun SM, Jiang SF, Xiong YW, Gong YS, Qiao H, Zhang WY, Wu Y (2018b) iTRAQ-based quantitative proteomic analysis of the androgenic glands of the oriental river prawn, Macrobrachium nipponense, during nonreproductive and reproductive seasons. Comp Biochem Physiol D 26:50–57Google Scholar
- Kang XJ, Liu ZM, Zhou KX, Wang SA (1998) Primary study of exogenous steroid hormone effect on the testis development of the Chinese Mitten-Handed Crab, Eriocheir sinensis (Crustcea, Decapoda). Donghai Mar Sci 16(1):39–44Google Scholar
- Li GL, Liu XC, Lin HR (2006) Effects of 17α-methyltestosterone on sex reversal in red-spotted grouper, Epinephelus akaara. J Fish China 30(2):145–150Google Scholar
- Li FJ, Bai HK, Xiong YW, Fu HT, Jiang SF, Jiang SF, Jin SB, Sun SM, Qiao H, Zhang WY (2015) Molecular characterization of insulin-like androgenic gland hormone-binding protein gene from the oriental river prawn Macrobrachium nipponense and investigation of its transcriptional relationship with the insulin-like androgenic gland hormone gene. Gen Comp Endocr 216(6):152–160CrossRefGoogle Scholar
- Lou YD, Song TF, Wang YM, Wei H, Wu MC, Xu HM, Wang ZQ (1994) Studies on sex control in allogynogenetic crucian carp. J Fish China 3:169–176Google Scholar
- Qiao H, Xiong YW, Zhang WY, Fu HT, Jiang SF, Sun SM, Bai HK, Jin SB, Gong YS (2015) Characterization, expression, and function analysis of gonad-inhibiting hormone in oriental river prawn, Macrobrachium nipponense and its induced expression by temperature. Comp Biochem Physiol A 185:1–8CrossRefGoogle Scholar
- Sagi A, Cohen D (1990) Growth, maturation and progeny of sex-reversed Macrobrachium rosenbetgii males. World Aquac 21(4):87–90Google Scholar
- Yu H, Miyake S (1972) Five species of the genus Macrobrachium (Crustacea, Decapoda, Palaemonidae) from Taiwan. OHMU 3:45–55Google Scholar
- Zhang PY, Hu W, Wang YP, Zhu ZY (2000) Methyltestosterone induction of sex reversal in allogynogenetic crucian carp. Hereditas 22(1):25–27Google Scholar