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

International Journal of Molecular Sciences, Год журнала: 2018, Номер 19(8), С. 2258 - 2258

Опубликована: Авг. 1, 2018

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

Язык: Английский

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Characterization of G-protein coupled receptors from the blackback land crab Gecarcinus lateralis Y organ transcriptome over the molt cycle

BMC Genomics, Год журнала: 2019, Номер 20(1)

Опубликована: Янв. 22, 2019

G-protein coupled receptors (GPCRs) are ancient, ubiquitous, constitute the largest family of transducing cell surface proteins, and integral to communication via an array ligands/neuropeptides. Molt inhibiting hormone (MIH) is a key neuropeptide that controls growth reproduction in crustaceans by regulating molt cycle. It inhibits ecdysone biosynthesis pair endocrine glands (Y-organs; YOs) through binding yet uncharacterized GPCR, which triggers signalling cascade, leading inhibition ecdysis sequence. When MIH release stops, synthesized released hemolymph. A peak titer followed molting event. transcriptome blackback land crab Gecarcinus lateralis YOs across was utilized this study curate list GPCRs their expression order better assess involved process.Ninety-nine G. putative were obtained screening YO against Pfam database. Phylogenetic analysis classified 49 as class (Rhodopsin-like receptor), 35 B (Secretin 9 C (metabotropic glutamate). Further phylogenetic identified GPCRs, including those for Allatostatin A, B, Bursicon, CCHamide, FMRFamide, Proctolin, Corazonin, Relaxin, biogenic amine Serotonin. Three clustered with recently CHH (CHHRs), differential over cycle suggests they associated ecdysteroidogenesis regulation. Two Corazonin showed much higher compared all other suggesting important role regulation.Molting requires orchestrated regulation ecdysteroid synthesis multiple neuropeptides. In study, we curated comprehensive expressed could include receptor whose activation negatively regulates molting. Orthologs found be insects also identified, LGR3 receptor, latter at level than receptors,

Язык: Английский

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

Comparative Biochemistry and Physiology Part B Biochemistry and Molecular Biology, Год журнала: 2018, Номер 218, С. 1 - 12

Опубликована: Янв. 5, 2018

Язык: Английский

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Transcriptome analysis of five ovarian stages reveals gonad maturation in female Macrobrachium nipponense

BMC Genomics, Год журнала: 2021, Номер 22(1)

Опубликована: Июль 6, 2021

Abstract Background Macrobrachium nipponense is an economically important species of freshwater shrimp in China. Unlike other marine shrimps, the ovaries adult female M. can mature rapidly and periodically during reproductive period, but resulting high stocking densities environmental deterioration negatively impact harvest yield economic benefits. To better understand ovary development , we performed systematic transcriptome sequencing five different stages ovarian maturation. Results We obtained 255,966 Gb quality data from 15 samples. Of 105,082 unigenes that were selected, 30,878 successfully annotated. From these unigenes, identified 17 differentially expressed genes three distinct gene expression patterns related to biological processes. found cathepins, legumains, cystatin enriched lysosome pathway, they are vitellogenin hydrolysis. Additionally, myosin heavy chain 67 participated oocyte excretion. Conclusions provide first detailed relating maturation cycle . Our results reference information about genomics, molecular biology, physiology, population genetics crustaceans. It conducive further solve problem rapid aspects energy supply cell division.

Язык: Английский

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A transcriptome study on Macrobrachium nipponense hepatopancreas experimentally challenged with white spot syndrome virus (WSSV)

PLoS ONE, Год журнала: 2018, Номер 13(7), С. e0200222 - e0200222

Опубликована: Июль 6, 2018

White spot syndrome virus (WSSV) is one of the most devastating pathogens cultured shrimp, responsible for massive loss its commercial products worldwide. The oriental river prawn Macrobrachium nipponense an economically important species that widely farmed in China and adult prawns can be infected by WSSV. However, molecular mechanisms host pathogen interaction remain unknown. There urgent need to learn between M. WSSV which will able offer a solution controlling spread Next Generation Sequencing (NGS) was used this study determin transcriptome differences comparison control WSSV-challenged moribund samples, survived samples hepatopancreas nipponense. A total 64,049 predicted unigenes were obtained classified into 63 functional groups. Approximately, 4,311 differential expression genes identified with 3,308 up-regulated when comparing control. In control, 1,960 764 up-regulated. contrast two libraries, 300 mutual DEGs 95 205 down-regulated genes. All performed GO KEGG analysis, overall 85 immune-related these gene groups 13 functions 4 pathways, such as protease inhibitors, heat shock proteins, oxidative stress, recognition immune receptors, PI3K/AKT/mTOR pathway, MAPK signaling pathway Ubiquitin Proteasome Pathway. Ten valuable responses against selected from those furture discuss response Results contribute better understanding WSSV, provide information identifying novel absence genome Furthermore, large number transcripts could strong basis future genomic research on

Язык: Английский

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Transcriptomic comparison between wild-caught and domesticated black tiger shrimp (Penaeus monodon) in early and late-vitellogenic broodstock females

Aquaculture Reports, Год журнала: 2025, Номер 41, С. 102675 - 102675

Опубликована: Фев. 13, 2025

Язык: Английский

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Deciphering the molecular regulatory mechanism orchestrating ovary development of the Pacific whiteleg shrimp Litopenaeus vannamei through integrated transcriptomic analysis of reproduction-related organs

Aquaculture, Год журнала: 2020, Номер 533, С. 736160 - 736160

Опубликована: Ноя. 18, 2020

Язык: Английский

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Guidelines for RNA-seq projects: applications and opportunities in non-model decapod crustacean species

Hydrobiologia, Год журнала: 2018, Номер 825(1), С. 5 - 27

Опубликована: Июль 3, 2018

Язык: Английский

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Neural Ganglia Transcriptome and Peptidome Associated with Sexual Maturation in Female Pacific Abalone (Haliotis discus hannai)

Genes, Год журнала: 2019, Номер 10(4), С. 268 - 268

Опубликована: Апрель 2, 2019

Genetic information of reproduction and growth is essential for sustainable molluscan fisheries aquaculture management. However, there limited knowledge regarding the reproductive activity commercially important Pacific abalone Haliotis discus hannai. We performed de novo transcriptome sequencing ganglia in sexually immature mature female to better understand sexual maturation process underlying molecular mechanisms. Of ~305 million high-quality clean reads, 76,684 transcripts were novo-assembled with an average length 741 bp, 28.54% which annotated classified according Gene Ontology terms. There 256 differentially expressed genes between abalone. Tandem mass spectrometry analysis, as compared predicted-peptide database unigenes, identified 42 neuropeptide precursors, including 29 validated by peptidomic analyses. Label-free quantification revealed differential occurrences 18 families abalone, achatin, FMRFamide, crustacean cardioactive peptide, pedal peptide A B that significantly more frequent at stage. These results represent first significant contribution both maturation-related transcriptomic resources provide insight into roles various neuropeptides regulation marine gastropods.

Язык: Английский

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Molecular cloning, expression pattern analysis, and in situ hybridization of a Transformer-2 gene in the oriental freshwater prawn, Macrobrachium nipponense (de Haan, 1849)

3 Biotech, Год журнала: 2019, Номер 9(6)

Опубликована: Май 7, 2019

Язык: Английский

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