Abstract
A critical aspect of real-time PCR is the presence of housekeeping genes (HKGs) as an internal control for the normalization of expression data for genes of interest. It is necessary to select correct HKGs in the investigation of various pathologies. Thereby, we analyzed the stability of expression of the HKGs in Parkinson’s disease (PD). The work was carried out in the peripheral blood of patients with PD and in the brain tissues and peripheral blood of mice with MPTP-induced PD. As a result, Aars was the most stably expressed HKG in the mouse brain as a whole. However, different genes were more stably expressed in different parts of the brain. Polr2f was the most stably expressed in the cortex, Psmd6 was the most stably expressed in the cerebellum, and Psmd7 was the most stably expressed in the striatum and substantia nigra. HKGs were different in similar tissues of the studied organisms. Polr2f was the most stably expressed HKG in the peripheral blood of mice, whereas PSMD6 was the most stably expressed gene in humans. Thus, there is no universal HKG both for different brain tissues of one organism and for similar tissues of different organisms. Furthermore, the identified most stably expressed HKGs can be considered as such only under conditions in PD.
Highlights
As the analysis shows, housekeeping genes (HKGs) in similar tissues of the studied organisms are different
Data from one expression study often do not reproduce the results of other similar studies; differences in the results obtained may be partly explained by the choice and instability of expression of HKGs in varying experimental conditions
The data obtained indicate that there is no universal HKG both for different brain tissues of one organism and for similar tissues of different organisms. This conclusion is supported by other studies: researchers must select and validate a set of reference genes for each specific cell line [41]
Summary
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. Active investigations of gene expression are being carried out. The simplest and most widely used method for analysis of gene expression at the mRNA level is a semiquantitative approach based on the reverse transcription (RT) reaction and realtime quantitative polymerase chain reaction (qPCR). This method is considered the gold standard for the study of changes in gene expression at the mRNA level in biomedical investigations [1,2]
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