污染物暴露下Mugilogobius chulae不同组织中qRT-PCR分析稳定参考基因的评估和选择
{"_":"Evaluation and Selection of Stable Reference Genes for qRT-PCR Analysis in Different Tissues of Under Pollutant Exposure.","i":["Mugilogobius chulae"]}
摘要(中文译文)
可靠的标准化对于准确定量实时PCR（qRT-PCR）分析至关重要，然而，尚未在一种新兴的用于生理学和环境研究的海洋实验鱼中系统评估合适的参考基因。本研究在性成熟雄性和雌性该鱼的五种组织（脑、鳃、性腺、肠道和肝脏）中，在溶剂对照（DMSO）、双酚A（BPA）、镉（Cd）和磺胺二甲嘧啶（SMX）处理条件下，评估了17个候选参考基因。采用比较ΔCt法、NormFinder、geNorm、BestKeeper以及综合RefFinder排名评估基因表达稳定性。结果显示，参考基因稳定性具有明显的组织特异性和分析方法依赖性。综合RefFinder分析鉴定出、、、和作为合并数据集中最稳定的基因，而、和则最不稳定。geNorm分析表明，脑、鳃、肠道和肝脏需要两个参考基因即可，而性腺需要四个，跨组织比较则需要六个。这些发现为该鱼中参考基因的选择提供了首个系统性依据，并为该物种未来的qRT-PCR研究奠定了重要的方法学基础，尤其是在内分泌干扰、生殖生理学和海洋生态毒理学研究中。
Abstract
Reliable normalization is essential for accurate quantitative real-time PCR (qRT-PCR) analysis, yet suitable reference genes have not been systematically evaluated in , an emerging marine experimental fish used in physiological and environmental research. In this study, 17 candidate reference genes were evaluated in five tissues (brain, gill, gonad, intestine, and liver) of sexually mature male and female under solvent control (DMSO), bisphenol A (BPA), cadmium (Cd), and sulfamethazine (SMX) treatment conditions. Gene-expression stability was assessed using the comparative ΔCt method, NormFinder, geNorm, BestKeeper, and the integrated RefFinder ranking. The results showed that reference-gene stability was strongly tissue-specific and analysis-dependent. The integrated RefFinder analysis identified , , , , and as the most stable genes in the pooled dataset, whereas , , and were the least stable. geNorm analysis indicated that two reference genes were sufficient for brain, gill, intestine, and liver, whereas four were required for the gonad and six for cross-tissue comparisons. These findings provide the first systematic basis for reference-gene selection in and establish an important methodological foundation for future qRT-PCR studies in this species, particularly in research on endocrine disruption, reproductive physiology, and marine ecotoxicology.
如何引用
AMA
Zhongdian Dong, Jiahao Gao, Xiaobin Li, Zhishan Chen, Jianjun Li, Jian Liao, et al. {"_":"Evaluation and Selection of Stable Reference Genes for qRT-PCR Analysis in Different Tissues of Under Pollutant Exposure.","i":["Mugilogobius chulae"]}. Animals : an open access journal from MDPI. 2026; doi:10.3390/ani16091412.
APA
Zhongdian Dong, Jiahao Gao, Xiaobin Li, Zhishan Chen, Jianjun Li, Jian Liao, et al (2026). {"_":"Evaluation and Selection of Stable Reference Genes for qRT-PCR Analysis in Different Tissues of Under Pollutant Exposure.","i":["Mugilogobius chulae"]}. Animals : an open access journal from MDPI. https://doi.org/10.3390/ani16091412
GB/T 7714
Zhongdian Dong, Jiahao Gao, Xiaobin Li, Zhishan Chen, Jianjun Li, Jian Liao, et al. {"_":"Evaluation and Selection of Stable Reference Genes for qRT-PCR Analysis in Different Tissues of Under Pollutant Exposure.","i":["Mugilogobius chulae"]}[J]. Animals : an open access journal from MDPI, 2026 doi:10.3390/ani16091412.
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