pH but not hypoxia affects neonatal gene expression: Relevance for housekeeping gene selection

Abstract

Objective. To identify a candidate neonatal housekeeping gene and to determine the effects of pH and PaO₂ on the stability of newborn gene expression in physiologically hypoxic and acidotic newborn blood.

Methods. Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) amplification was performed for four commonly used housekeeping genes (GAPDH, β-actin, cyclophilin, 28S rRNA) on extracted RNA. Blood gas analyses determined pH and PaO₂ levels.

Results and conclusions. β-Actin was the least variable and GAPDH the most variable housekeeping gene studied. pH negatively correlated with gene expression levels. PaO₂ levels did not significantly affect gene expression. These results inform selection of housekeeping genes for neonatal mRNA research.

Keywords:

• Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR)
• housekeeping gene
• newborn gene expression