2014
DOI: 10.1371/journal.pone.0109902
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Validation of Housekeeping Genes in the Brains of Rats Submitted to Chronic Intermittent Hypoxia, a Sleep Apnea Model

Abstract: Obstructive sleep apnea (OSA) is a syndrome characterized by intermittent nocturnal hypoxia, sleep fragmentation, hypercapnia and respiratory effort, and it has been associated with several complications, such as diabetes, hypertension and obesity. Quantitative real-time PCR has been performed in previous OSA-related studies; however, these studies were not validated using proper reference genes. We have examined the effects of chronic intermittent hypoxia (CIH), which is an experimental model mainly of cardio… Show more

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Cited by 47 publications
(40 citation statements)
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“…This is in contrast to previous studies testing across different rat tissues (Svingen et al, 2015) or brain tissue from rats (Cook et al, 2009;Yang et al, 2008;Zhou et al, 2016), mice (Crans et al, 2019) or humans (Rydbirk et al, 2016). Notably, brain tissue from a rat sleep apnea model displayed high stability of all tested RGs except 18S rRNA (Julian et al, 2014), a stability also seen in rat brains across fetal developmental stages (Al-Bader & Al-Sarraf, 2005), which agrees with our findings of relative high stability among RGs in this tissue (Julian et al, 2014). There are, however, studies showing unstable expression of common RGs depending on experimental parameters and what brain regions are examined (Cook et al, 2009;Swijsen et al, 2012;Yang et al, 2008;Zhou et al, 2016).…”
Section: Discussioncontrasting
confidence: 90%
“…This is in contrast to previous studies testing across different rat tissues (Svingen et al, 2015) or brain tissue from rats (Cook et al, 2009;Yang et al, 2008;Zhou et al, 2016), mice (Crans et al, 2019) or humans (Rydbirk et al, 2016). Notably, brain tissue from a rat sleep apnea model displayed high stability of all tested RGs except 18S rRNA (Julian et al, 2014), a stability also seen in rat brains across fetal developmental stages (Al-Bader & Al-Sarraf, 2005), which agrees with our findings of relative high stability among RGs in this tissue (Julian et al, 2014). There are, however, studies showing unstable expression of common RGs depending on experimental parameters and what brain regions are examined (Cook et al, 2009;Swijsen et al, 2012;Yang et al, 2008;Zhou et al, 2016).…”
Section: Discussioncontrasting
confidence: 90%
“…We quantified the gene expression using the Comparative threshold (Ct) method (ΔΔCt method) [18,19], and the amount of target gene was normalized to ACTB as housekeeping gene (as it was stable across all samples) and determined by 2 −ΔΔCt , as previously described [20,21], with relative expression levels reported as fold change. Ct values higher than the cut-off of 35 were not considered as a reliable expression value, according to the manufacturer's recommendations, and therefore were excluded from the statistical analysis.…”
Section: Gene Expression Analysismentioning
confidence: 99%
“…This difference may be attributed to different mathematical models used in each program [24]. On the other hand, there was found similar results for the software geNorm and Normfinder but not for BestKeeper, this can be justified by the fact that this software used the Pearson correlation method to classify the reference genes, a different method compared to the others software [3].…”
Section: Genorm Analysismentioning
confidence: 91%
“…Our group has validated reference genes for rat models of sleep deprivation [2] and hypoxia [3]. To our knowledge, there are no studies concerning validation of reference genes to analyze the effects of resistance exercise (REx) and androgenic anabolic steroids (AAS) use on gene expression.…”
Section: Introductionmentioning
confidence: 99%