pH but not hypoxia affects neonatal gene expression: relevance for housekeeping gene selection.
OBJECTIVE
To identify a candidate neonatal housekeeping gene and to determine the effects of pH and PaO(2) 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, beta-actin, cyclophilin, 28S rRNA) on extracted RNA. Blood gas analyses determined pH and PaO(2) levels.


CONCLUSIONS
Beta-Actin was the least variable and GAPDH the most variable housekeeping gene studied. pH negatively correlated with gene expression levels. PaO(2) levels did not significantly affect gene expression. These results inform selection of housekeeping genes for neonatal mRNA research.
Maron
Jill L
JL
Department of Pediatrics, Tufts Medical Center, Boston, MA 02111, USA. jmaron@tufts-nemc.org
Arya
Michelle A
MA
Seefeld
Kimberly J
KJ
Peter
Inga
I
Bianchi
Diana W
DW
Johnson
Kirby L
KL
D000136:Acid-Base Equilibrium / Q000235:genetics*; D000199:Actins / Q000235:genetics; D002585:Cesarean Section; D021983:Cyclophilins / Q000235:genetics; D005260:Female; D005312:Fetal Blood; D020869:Gene Expression Profiling; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D006801:Humans; D000860:Hypoxia / Q000235:genetics*; D007231:Infant, Newborn; D011247:Pregnancy; D012339:RNA, Ribosomal, 28S / Q000235:genetics
D000199:Actins; D012339:RNA, Ribosomal, 28S; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D021983:Cyclophilins