Characterization of regulators of G-protein signaling RGS4 and RGS10 proteins in the postmortem human brain.
Regulator of G-protein signaling (RGS) proteins are a large family of proteins that accelerate GTPase rate of the Gα subunits and therefore, negatively regulate G-protein signaling. Expression of RGS4 and RGS10 proteins was characterized in human prefrontal cortex attending to methodological (subcellular localization, antibody specificity and sensitivity, postmortem delay (PMD) and storage conditions of the samples) and demographic issues (age and gender of the subjects). Anti-RGS4 (N-16) antibody revealed a unique and specific band of 38 kDa that was highly enriched in the plasma membrane. Anti-RGS10 (C-20) antibody revealed two specific bands of 24 and 27kDa, corresponding to two possible isoforms of this protein, which were predominantly localized in the cytosol. Antibody dilution and protein linearity studies confirmed the sensitivity of the signal. A large number of samples from 58 individuals presenting well spread PMD, storage time, age of the subjects at the time of death, and male and female distribution were studied. A positive linear relationship between the age and RGS4 immunoreactivity was observed. There was a negative influence of PMD on the RGS10 27 kDa band immunoreactivity but a positive relationship emerged between the PMD and RGS4 immunoreactivity. Storage time of the samples did not have any influence on RGS4 nor on RGS10 immunoreactivity, showing their stability at -70°C. When studying the RGS4 and RGS10 protein expression density in males and females, no significant difference was found between groups. This study demonstrates that RGS4 and RGS10 proteins can be detected by immunoreactive techniques in postmortem human brain cortex. The study provides important matching conditions that should be taken into account in postmortem brain studies of neuropsychiatric diseases.
Rivero
Guadalupe
G
Department of Pharmacology, University of the Basque Country, Leioa, Bizkaia, Spain.
Gabilondo
Ane M
AM
García-Sevilla
Jesús A
JA
La Harpe
Romano
R
Morentín
Benito
B
Javier Meana
J
J
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D005260:Female; D019204:GTP-Binding Proteins / Q000502:physiology; D006801:Humans; D008297:Male; D008875:Middle Aged; D011180:Postmortem Changes; D017397:Prefrontal Cortex / Q000378:metabolism* / Q000473:pathology; D020710:RGS Proteins / Q000502:physiology*; D015398:Signal Transduction / Q000502:physiology*; D055815:Young Adult
D020710:RGS Proteins; C101113:RGS10 protein, human; C098799:RGS4 protein; D019204:GTP-Binding Proteins