Identification and characterization of opsin gene and its role in ovarian maturation in the oriental river prawn Macrobrachium nipponense

Li, Fēi; Qiao, Hui; Fu, Hongtuo; Sun, Shengming; Zhang, Wenyi; Jin, Shubo; Jiang, Sufei; Gong, Yongsheng; Xiong, Yiwei; Wu, Yan; Hu, Yu‐Ning; Shan, Dongyan

doi:10.1016/j.cbpb.2017.12.016

Cited by 53 publications

(74 citation statements)

References 62 publications

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“…We confirmed the selected reference genes and verified their rankings based on qPCR by RNAi, which could reliably decrease the expression level of the target gene in M. nipponense [29,30]. The SST expression in the androgenic glands in M. nipponense was predicted to decrease by >90% over seven days after RNAi.…”

Section: Resultsmentioning

confidence: 57%

“…Previous researchers in our lab have proved that RNAi is able to significantly reduce the expression of specific genes in M. nipponense [29,30]. The SST gene is specifically expressed in androgenic glands [39], and its expression was predicted to decrease by >90% by seven days after the injection.…”

Section: Discussionmentioning

confidence: 99%

“…The protein prediction was performed using the open reading frame (ORF) finder (). Multiple alignments of amino acid sequences’ encoding were created using DNAMAN 6.0 [30]. …”

Section: Methodsmentioning

confidence: 99%

“…This study was approved by the Institutional Animal Care and Use Ethics Committee of the Freshwater Fisheries Research Center, the Chinese Academy of Fishery Sciences (Wuxi, China, FFRC125, 26 August 2016). All of the prawns were obtained from the Freshwater Fisheries Research Center, the Chinese Academy of Fishery Sciences, Wuxi, China (120°13′44″ E, 31°28′22″ N) [30]. …”

Section: Methodsmentioning

confidence: 99%

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Validation and Evaluation of Reference Genes for Quantitative Real-Time PCR in Macrobrachium Nipponense

Qiao

et al. 2018

IJMS

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Quantitative real-time PCR (qPCR) is widely used in molecular biology, although the accuracy of the quantitative results is determined by the stability of the reference genes used. Recent studies have investigated suitable reference genes for some crustaceans under various conditions, but studies in Macrobrachium nipponense are currently lacking. In this study, we selected the following seven genes from among 35 commonly used housekeeping genes as candidate qPCR reference genes for temporal and spatial expression: EIF (eukaryotic translation initiation factor 5A), 18S (18S ribosomal RNA), EF-1α (elongation factor-1α), GAPDH (glyceraldehyde-3-phosphate dehydrogenase), TUB (α-tubulin), β-act (β-actin), and RPL18 (Ribosomal protein L18). The stability of each reference gene was evaluated by GeNorm, NormFinder, BestKeeper, and comparative ∆C t methods, and was comprehensively ranked using RefFinder. RPL18 was shown to be the most suitable reference gene for adult M. nipponense tissues, while EIF was the most stable in different ovarian and embryo stages and in white spot syndrome virus infection, and β-act was the most stable reference gene under hypoxia stress. The reliability of the rankings was confirmed by RNA interference experiments. To the best of our knowledge, this represents the first systematic analysis of reference genes for qPCR experiments in M. nipponense, and the results will provide invaluable information for future research in closely related crustaceans.

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Section: Resultsmentioning

confidence: 57%

Section: Discussionmentioning

confidence: 99%

“…The protein prediction was performed using the open reading frame (ORF) finder (). Multiple alignments of amino acid sequences’ encoding were created using DNAMAN 6.0 [30]. …”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Validation and Evaluation of Reference Genes for Quantitative Real-Time PCR in Macrobrachium Nipponense

Qiao

et al. 2018

IJMS

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“…Hepatopancreas and gill samples were collected according to the requirements of the chromogenic in situ hybridization (CISH) technique. Fixed paraffin-embedded sections, standard deparaffinization, sectioning and staining were prepared as previously reported by our laboratory [30]. Slides were evaluated using a light microscope.…”

Section: In Situ Hybridization (Ish)mentioning

confidence: 99%

Molecular cloning, expression, and in situ hybridization analysis of MnGPx-3 and MnGPx-4 from oriental river prawn, Macrobrachium nipponense, in response to hypoxia and reoxygenation

Yang

et al. 2020

PLoS ONE

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Glutathione peroxidase (GPx) has been the focus of increased research because of its important role as an antioxidant and in reactive oxygen species (ROS) induced damage repair. Studies on GPxs have relevance with Macrobrachium nipponense because it has poor tolerance to hypoxia in Macrobrachium nipponense. The two subunits named as MnGPx-3 and MnGPx-4 according to the glutathione peroxidase nomenclature system. Both full-length cDNAs were cloned from the hepatopancreas. In this study, we analyzed the expression of two GPxs in Macrobrachium nipponense in response to changes in environmental oxygen. Expression levels of MnGPx-3 and MnGPx-4 indicated that both have strong responses to hypoxia. In situ hybridization showed that MnGPx-3 and MnGPx-4 were located in secretory and storage cells in hepatopancreas. These results suggest that GPx gene is expressed and released by secretory cells and released response to hypoxia. In the gill tissue, however, GPxs are located in blood cells, suggesting that they perform different functions in different tissues or organs. The results of in situ hybridization were consistent with those of quantitative Real-time PCR. This study provides a basis for understanding the oxidative stress response in M. nipponense under hypoxia.

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RNA interference analysis of the potential functions of cyclin-dependent kinase 2 in sexual reproduction of male oriental river prawns (Macrobrachium nipponense)

et al. 2023

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Identification and characterization of opsin gene and its role in ovarian maturation in the oriental river prawn Macrobrachium nipponense

Cited by 53 publications

References 62 publications

Validation and Evaluation of Reference Genes for Quantitative Real-Time PCR in Macrobrachium Nipponense

Validation and Evaluation of Reference Genes for Quantitative Real-Time PCR in Macrobrachium Nipponense

Molecular cloning, expression, and in situ hybridization analysis of MnGPx-3 and MnGPx-4 from oriental river prawn, Macrobrachium nipponense, in response to hypoxia and reoxygenation

RNA interference analysis of the potential functions of cyclin-dependent kinase 2 in sexual reproduction of male oriental river prawns (Macrobrachium nipponense)

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