Cathepsin D Plays a Vital Role in Macrobrachium nipponense of Ovary Maturation: Identification, Characterization, and Function Analysis

Dan Cheng; Wenyi Zhang; Sufei Jiang; Yiwei Xiong; Shubo Jin; Fangyan Pan; Junpeng Zhu; Yongsheng Gong; Yan Wu; Hui Qiao; Hongtuo Fu

doi:10.3390/genes13081495

Cathepsin D Plays a Vital Role in Macrobrachium nipponense of Ovary Maturation: Identification, Characterization, and Function Analysis

Genes (Basel). 2022 Aug 21;13(8):1495. doi: 10.3390/genes13081495.

Authors

Dan Cheng¹, Wenyi Zhang², Sufei Jiang², Yiwei Xiong², Shubo Jin^{1

2}, Fangyan Pan³, Junpeng Zhu¹, Yongsheng Gong², Yan Wu², Hui Qiao^{1

2}, Hongtuo Fu^{1

2

3}

Affiliations

¹ Wuxi Fisheries College, Nanjing Agricultural University, Wuxi 214081, China.
² Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture and Rural Affairs, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi 214081, China.
³ National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai 201306, China.

Abstract

The oriental river prawn Macrobrachium nipponense is an economically important aquacultural species. However, its aquaculture is negatively impacted by the rapid sexual maturation of female M. nipponense. The fast sexual maturation produces a large number of offspring which leads to a reduction in resilience, a low survival rate, and an increased risk of hypoxia, this in turn, seriously affects the economic benefits of prawn farming. Cathepsin D is a lysosomal protease involved in the ovarian maturation of M. nipponense. In the current study, the cDNA of the gene encoding cathepsin D (Mn-CTSD) was cloned from M. nipponense. The total length was 2391 bp and consisted of an open reading frame (ORF) of 1158 bp encoding 385 amino acids. Sequence analysis confirmed the presence of conserved N-glycosylation sites and characteristic sequences of nondigestive cathepsin D. The qPCR analysis indicated that Mn-CTSD was highly expressed in all tissues tested, most significantly in the ovaries, whereas in situ hybridization showed that expression occurred mainly in oocyte nuclei. Analysis of its expression during development showed that Mn-CTSD peaked during the O-IV stage of ovarian maturation. For the RNAi interference experiment, female M. nipponense specimens in the ovary stage I were selected. Injection of Mn-CTSD double-stranded (ds)RNA into female M. nipponense decreased the expression of Mn-CTSD in the ovaries, such that the Gonad Somatic Index (GSI) of the experimental group was significantly lower than that of the control group (1.79% versus 4.57%; p < 0.05). Ovary development reached the O-III stage in 80% of the control group, compared with 0% in the experimental group. These results suggest that Mn-CTSD dsRNA inhibits ovarian maturation in M. nipponense, highlighting its important role in ovarian maturation in this species and suggesting an approach to controlling ovarian maturation during M. nipponense aquaculture.

Keywords: Macrobrachium nipponense; RNAi; cathepsin D; mRNA expression; ovary maturation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Animals
Base Sequence
Cathepsin D / genetics
Cathepsin D / metabolism
Female
Ovary / metabolism
Palaemonidae* / genetics
Palaemonidae* / metabolism
RNA, Double-Stranded / metabolism

Substances

RNA, Double-Stranded
Cathepsin D

Grants and funding

This study was supported by the grants from Central Public-interest Scientific Institution Basal Research Fund CAFS (2020TD36); Jiangsu Agricultural Industry Technology System; the ear-marked fund for CARS-48 (CARS-48); the New cultivar breeding Major Project of Jiangsu province (PZCZ201745); the seed industry revitalization project of Jiangsu province (JBGS [2021] 118); the National Key R&D Program of China (2018YFD0901303). Thanks for the Jiangsu Province Platform for the Conservation and Utilization of Agricultural Germplasm.