title: There is no correlation between glucocorticoid receptor mRNA expression and protein binding in the brains of house sparrows (Passer domesticus).
abstract: The stress response represents an animal's attempt to cope with a noxious stimulus through a rapid release of corticosterone or cortisol (CORT) into the bloodstream, resulting in a suite of physiological and behavioral changes. These changes are mediated in large part through CORT's binding to two different intracellular receptors, the high-affinity mineralocorticoid receptor (MR) and the lower-affinity glucocorticoid receptor (GR). We tested the hypothesis that GR and MR mRNA expression would correlate with functional protein expression in neuronal tissue of wild-caught house sparrows (Passer domesticus). To test this hypothesis, we performed a parallel procedure in which protein concentrations were quantified in one half of house sparrow brains (n=16) using radioligand binding assays, and mRNA levels were quantified in the other brain half using reverse-transcriptase quantitative PCR (RT-qPCR). Two reference genes, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and TATA-box binding protein (TBP), were used for relative quantification of GR and MR mRNA. Quantifications showed that these two reference genes were not correlated with each other. Furthermore, there was no correlation between mRNA and protein levels for GR or MR using either reference gene, suggesting that regulation of mRNA and protein levels for MR and GR is not tightly linked. This study provides insight into the importance of regulatory steps between mRNA expression and the creation and stability of a functional protein. The overall conclusion is that mRNA expression cannot be used as a proxy for GR or MR binding in house sparrows.
authors.affiliation: Department of Biology, Tufts University, Medford, MA 02155, USA.
authors.forename: Carlos O
authors.initials: CO
authors.lastname: Medina
authors.forename: Christine R
authors.initials: CR
authors.lastname: Lattin
authors.forename: M
authors.initials: M
authors.lastname: McVey
authors.forename: L Michael
authors.initials: LM
authors.lastname: Romero
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism*; D005786:Gene Expression Regulation; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D012333:RNA, Messenger / Q000235:genetics*; D011965:Receptors, Glucocorticoid / Q000235:genetics* / Q000378:metabolism*; D018161:Receptors, Mineralocorticoid / Q000235:genetics* / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D046370:Sparrows / Q000235:genetics* / Q000378:metabolism*
chemical_list: D012333:RNA, Messenger; D011965:Receptors, Glucocorticoid; D018161:Receptors, Mineralocorticoid; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases
keywords: Glucocorticoid receptor; Glyceraldehyde-3-phosphate dehydrogenase; Mineralocorticoid receptor; Reverse transcriptase quantitative PCR; TATA-box binding protein