Reference genes for quantitative PCR in the adipose tissue of mice with metabolic disease
| Autor: | Priscila Ausina, David Majerowicz, Fernanda Almeida-Oliveira, João G. B. Leandro, Mauro Sola-Penna |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
Male
0301 basic medicine Mice Obese Adipose tissue Computational biology Biology Real-Time Polymerase Chain Reaction Diabetes Mellitus Experimental Mice 03 medical and health sciences 0302 clinical medicine Metabolic Diseases Reference genes Diabetes mellitus Gene expression medicine Animals Gene Pharmacology Messenger RNA General Medicine Reference Standards medicine.disease Molecular biology Obesity 030104 developmental biology Real-time polymerase chain reaction Adipose Tissue Gene Expression Regulation 030217 neurology & neurosurgery |
| Zdroj: | Biomedicine & Pharmacotherapy. 88:948-955 |
| ISSN: | 0753-3322 |
| DOI: | 10.1016/j.biopha.2017.01.091 |
| Popis: | Obesity and diabetes are metabolic diseases and they are increasing in prevalence. The dynamics of gene expression associated with these diseases is fundamental to identifying genes involved in related biological processes. qPCR is a sensitive technique for mRNA quantification and the most commonly used method in gene-expression studies. However, the reliability of these results is directly influenced by data normalization. As reference genes are the major normalization method used, this work aims to identify reference genes for qPCR in adipose tissues of mice with type-I diabetes or obesity. We selected 12 genes that are commonly used as reference genes. The expression of these genes in the adipose tissues of mice was analyzed in the context of three different experimental protocols: 1) untreated animals; 2) high-fat-diet animals; and 3) streptozotocin-treated animals. Gene-expression stability was analyzed using four different algorithms. Our data indicate that TATA-binding protein is stably expressed across adipose tissues in control animals. This gene was also a useful reference when the brown adipose tissues of control and obese mice were analyzed. The mitochondrial ATP synthase F1 complex gene exhibits stable expression in subcutaneous and perigonadal adipose tissue from control and obese mice. Moreover, this gene is the best reference for qPCR normalization in adipose tissue from streptozotocin-treated animals. These results show that there is no perfect stable gene suited for use under all experimental conditions. In conclusion, the selection of appropriate genes is a prerequisite to ensure qPCR reliability and must be performed separately for different experimental protocols. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect