Expression profiles of CHH2 gene in redclaw crayfish Cherax quadricarinatus and its role in ovarian development

doi:10.3969/j.issn.1004-1524.2023.01.04

Acta Agriculturae Zhejiangensis ›› 2023, Vol. 35 ›› Issue (1): 33-40.DOI: 10.3969/j.issn.1004-1524.2023.01.04

• Animal Science • Previous Articles Next Articles

Expression profiles of CHH2 gene in redclaw crayfish Cherax quadricarinatus and its role in ovarian development

CHEN Leran¹^,²(), ZHENG Jianbo², JIA Yongyi², CHI Meili², LI Fei², CHENG Shun², LIU Shili², LIU Yinuo², JIANG Wenping², GU Zhimin¹^,^*()

1. Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture and Rural Affairs, College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China
2. Key Laboratory of Healthy Freshwater Aquaculture, Ministry of Agriculture and Rural Affairs, Key Laboratory of Freshwater Aquaculture Genetic and Breeding of Zhejiang Province, Zhejiang Institute of Freshwater Fisheries, Huzhou 313001, China

Received:2022-04-06 Online:2023-01-25 Published:2023-02-21

Abstract

Abstract:

To study the role of crustacean hyperglycemic hormone gene (CHH) in the ovarian development of Cherax quadricarinatus, sequence of CHH2 gene was identified based on transcriptomic data, and the basic characteristics were analyzed by bioinformatic. qRT-PCR was used to detect the expression level of CHH2 in different tissues and ovarian development stages. The results showed that CHH proteins were highly conserved among crustacean species, and they had high homologies with 66.84%. The tissue distribution at transcriptional level displayed that CHH2 was hardly to be detected in hepatopancreas, and was detected at low level in muscle, gills, heart and intestine, while highly expressed in ovary. In addition, CHH2 was expressed in all stages of ovarian development and exhibited an abnormal expression pattern with the lowest expression level at mature stage. On the other hand, CHH2 expression was significantly reduced in ovarian tissues after ESA treatment and E2 injection. Furthermore, VTG was regarded as an important marker for ovarian development, VTG-mRNA appeared to be increased through RNAi-induced gene silencing. In conclusion, CHH2 might negatively regulate ovarian development, and this study would provide new insights for further exploring the mechanism of crustacean ovarian development.

Key words: Cherax quadricarinatus, crustacean hyperglycemic hormone 2 (CHH2), RNAi, expression profile, ovarian development

CLC Number:

S917.4

CHEN Leran, ZHENG Jianbo, JIA Yongyi, CHI Meili, LI Fei, CHENG Shun, LIU Shili, LIU Yinuo, JIANG Wenping, GU Zhimin. Expression profiles of CHH2 gene in redclaw crayfish Cherax quadricarinatus and its role in ovarian development[J]. Acta Agriculturae Zhejiangensis, 2023, 35(1): 33-40.

Figures/Tables 8

References 27

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蛋白质名称 Protein name	物种 Species	基因登录号 GenBank No.	蛋白质名称 Protein name	物种 Species	基因登录号 GenBank No.
CHH	马氏沼虾Macrobrachium malcolmsonii	QIX07539.1	CHH-B	挪威龙虾Nephrops norvegicus	AAQ22392.1
CHH	北极甜虾Pandalus japonicus	AFG16933.1	CHH	窄指河螯虾Astacus leptodactylus	AAX09331.1
CHH-B	欧洲龙虾Homarus gammarus	ABA42180.1	CHH	美洲螯龙虾Homarus americanus	XP_042231195.1
CHH2	大西洋无眼裂缝虾Rimicaris kairei	ACS35347.1	CHH	细纹方蟹Grapsus tenuicrustatus	AER27832.1
CHH2	红螯螯虾Cherax quadricarinatus	MH210971.1	CHH-B	南美白对虾Penaeus vannamei	AAN86055.1
CHH	云斑厚纹蟹Pachygrapsus marmoratus	AAM21927.1	CHH3	红螯螯虾Cherax quadricarinatus	MH210972.1
CHH1	红螯螯虾Cherax quadricarinatus	MH210970.1

蛋白质名称 Protein name	物种 Species	基因登录号 GenBank No.	蛋白质名称 Protein name	物种 Species	基因登录号 GenBank No.
CHH	马氏沼虾Macrobrachium malcolmsonii	QIX07539.1	CHH-B	挪威龙虾Nephrops norvegicus	AAQ22392.1
CHH	北极甜虾Pandalus japonicus	AFG16933.1	CHH	窄指河螯虾Astacus leptodactylus	AAX09331.1
CHH-B	欧洲龙虾Homarus gammarus	ABA42180.1	CHH	美洲螯龙虾Homarus americanus	XP_042231195.1
CHH2	大西洋无眼裂缝虾Rimicaris kairei	ACS35347.1	CHH	细纹方蟹Grapsus tenuicrustatus	AER27832.1
CHH2	红螯螯虾Cherax quadricarinatus	MH210971.1	CHH-B	南美白对虾Penaeus vannamei	AAN86055.1
CHH	云斑厚纹蟹Pachygrapsus marmoratus	AAM21927.1	CHH3	红螯螯虾Cherax quadricarinatus	MH210972.1
CHH1	红螯螯虾Cherax quadricarinatus	MH210970.1

引物名称 Primer name	序列 Sequences (5'→3')	目的 Purpose
CHH2-RT-F	CCTCTCACAAGATCACAGCCTCAAC	qRT-PCR
CHH2-RT-R	GTAACAATCATCACAAACTCGGTCAAGC	qRT-PCR
β-actin-RT-F	TCACTGCTCTGGCTCCTGCTAC	qRT-PCR
β-actin-RT-R	ACTCGTCGTACTCCTCCTTGGTG	qRT-PCR
VTG-RT-F	CAGTCCGAGTATTCCGCCAAGAAC	qRT-PCR
VTG-RT-R	AACCAGTCGTTACCAACTTCACCATC	qRT-PCR
CHH2-dsRNA-F	TAATACGACTCACTATAGGGatgacctcctgcagaacgat	RNA interference
CHH2-dsRNA-R	TAATACGACTCACTATAGGGacagtaacatggtgttccga	RNA interference

引物名称 Primer name	序列 Sequences (5'→3')	目的 Purpose
CHH2-RT-F	CCTCTCACAAGATCACAGCCTCAAC	qRT-PCR
CHH2-RT-R	GTAACAATCATCACAAACTCGGTCAAGC	qRT-PCR
β-actin-RT-F	TCACTGCTCTGGCTCCTGCTAC	qRT-PCR
β-actin-RT-R	ACTCGTCGTACTCCTCCTTGGTG	qRT-PCR
VTG-RT-F	CAGTCCGAGTATTCCGCCAAGAAC	qRT-PCR
VTG-RT-R	AACCAGTCGTTACCAACTTCACCATC	qRT-PCR
CHH2-dsRNA-F	TAATACGACTCACTATAGGGatgacctcctgcagaacgat	RNA interference
CHH2-dsRNA-R	TAATACGACTCACTATAGGGacagtaacatggtgttccga	RNA interference

Expression profiles of CHH2 gene in redclaw crayfish Cherax quadricarinatus and its role in ovarian development

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