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 cardiovascular consequences of OSA, on reference genes, including beta-actin, beta-2-microglobulin, glyceraldehyde-3-phosphate dehydrogenase, hypoxanthine guanine phosphoribosyl transferase and eukaryotic 18S rRNA, in different areas of the brain. All stability analyses were performed using the geNorm, Normfinder and BestKeeper software programs. With exception of the 18S rRNA, all of the evaluated genes were shown to be stable following CIH exposure. However, gene stability rankings were dependent on the area of the brain that was analyzed and varied according to the software that was used. This study demonstrated that CIH affects various brain structures differently. With the exception of the 18S rRNA, all of the tested genes are suitable for use as housekeeping genes in expression analyses.
Highlights
Obstructive sleep apnea (OSA) is the most common sleeprelated breathing disorder
Many animal models that are commonly been used in the study of hypoxia have been developed over the years, of which the most widely used is the chronic intermittent hypoxia (CIH) model, which simulates only one factor of OSA
All of the samples were initially treated with DNase I, and their integrity was evaluated by the assessment of intact 28S and 18S rRNAs on agarose gels
Summary
Obstructive sleep apnea (OSA) is the most common sleeprelated breathing disorder. OSA has been associated with several comorbidities, such as cardiovascular disease, hypertension, diabetes, cognitive impairments and metabolic syndrome [1,2,3,4,5,6,7]. CIH, occurring isolated or in association with sleep fragmentation, has been demonstrated to lead to several changes that are similar to those found in individuals with OSA, such as cognitive impairment, insulin resistance and hypertension [1,9,10,11,12]. It has been demonstrated that CIH acts by modulating presympathetic neurons activity in the rostral ventrolateral medulla [14], increasing sympathetic activity. These data suggest that intermittent hypoxia plays an important role in OSA. CIH model lacks of several OSA factors, it is still an important tool to study OSA, mainly in cardiovascular area
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