小鼠棕色脂肪细胞分化和体外产热参考基因的比较评估

IF 3.5 4区 生物学 Q2 ENDOCRINOLOGY & METABOLISM
Adipocyte Pub Date : 2024-12-01 Epub Date: 2024-03-25 DOI:10.1080/21623945.2024.2330355
Trang Huyen Lai, Jin Seok Hwang, Quang Nhat Ngo, Dong-Kun Lee, Hyun Joon Kim, Deok Ryong Kim
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引用次数: 0

摘要

棕色脂肪组织(BAT)的成脂分化和产热过程是动态的,会改变表型和基因表达。基因表达分析中适当的参考基因对于减少实验差异和确保 PCR 的有效性至关重要。不可靠的参考基因会导致错误的基因表达定量,造成数据误读。本研究的重点是利用 Ucp1-luciferase ThermoMouse 模型中的棕色脂肪细胞,为小鼠棕色脂肪细胞研究确定合适的参考基因。通过各种算法(包括 DeltaCq、BestKeeper、geNorm、Normfinder 和 RefFinder)验证了 13 个管家基因,并对脂肪生成和产热条件下的基因表达数据进行了比较分析。Tbp和Rer1成为棕色脂肪生成过程中Ucp1和Pparg表达的最佳参照物,而Tbp和Ubc则是热生成过程中这些靶基因表达分析的理想参照物。相反,某些传统参考基因,包括 Actb、Tubb5 和 Gapdh,在这两种条件下作为参考基因都不稳定。这些发现强调,在特定生物系统中进行基因表达分析时,参考基因的选择是确保得出准确结论的关键因素。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
A comparative assessment of reference genes in mouse brown adipocyte differentiation and thermogenesis in vitro.

Adipogenic differentiation and thermogenesis in brown adipose tissue (BAT) undergo dynamic processes, altering phenotypes and gene expressions. Proper reference genes in gene expression analysis are crucial to mitigate experimental variances and ensure PCR efficacy. Unreliable reference genes can lead to erroneous gene expression quantification, resulting in data misinterpretation. This study focused on identifying suitable reference genes for mouse brown adipocyte research, utilizing brown adipocytes from the Ucp1-luciferase ThermoMouse model. Comparative analysis of gene expression data under adipogenesis and thermogenesis conditions was conducted, validating 13 housekeeping genes through various algorithms, including DeltaCq, BestKeeper, geNorm, Normfinder, and RefFinder. Tbp and Rer1 emerged as optimal references for Ucp1 and Pparg expression in brown adipogenesis, while Tbp and Ubc were ideal for the expression analysis of these target genes in thermogenesis. Conversely, certain conventional references, including Actb, Tubb5, and Gapdh, proved unstable as reference genes under both conditions. These findings stress the critical consideration of reference gene selection in gene expression analysis within specific biological systems to ensure accurate conclusions.

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来源期刊
Adipocyte
Adipocyte Medicine-Histology
CiteScore
6.50
自引率
3.00%
发文量
46
审稿时长
32 weeks
期刊介绍: Adipocyte recognizes that the adipose tissue is the largest endocrine organ in the body, and explores the link between dysfunctional adipose tissue and the growing number of chronic diseases including diabetes, hypertension, cardiovascular disease and cancer. Historically, the primary function of the adipose tissue was limited to energy storage and thermoregulation. However, a plethora of research over the past 3 decades has recognized the dynamic role of the adipose tissue and its contribution to a variety of physiological processes including reproduction, angiogenesis, apoptosis, inflammation, blood pressure, coagulation, fibrinolysis, immunity and general metabolic homeostasis. The field of Adipose Tissue research has grown tremendously, and Adipocyte is the first international peer-reviewed journal of its kind providing a multi-disciplinary forum for research focusing exclusively on all aspects of adipose tissue physiology and pathophysiology. Adipocyte accepts high-profile submissions in basic, translational and clinical research.
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