GABA(A) alpha1 and alpha2 receptor subunit expression in rostral ventrolateral medulla in nonpregnant and pregnant rats.
Pregnancy results in attenuated baroreflex mediated sympathoexcitatory responses which may be due to potentiation of gamma-aminobutyric acid (GABA) inhibition in the rostral ventrolateral medulla (RVLM). The major metabolite of progesterone, 3alpha-hydroxy-dihydroprogesterone (3alpha-OH-DHP), which is elevated in pregnancy, is a potent neurosteroid positive modulator of GABA(A) receptors, and sensitivity of GABA(A) receptors to 3alpha-OH-DHP is dependent on the receptor subunit composition. The purpose of this study was to evaluate the GABA(A) alpha(1) and alpha(2) receptor subunit mRNA and protein expression in the RVLM of nonpregnant and late term pregnant rats. Micropunches of RVLM were collected from nonpregnant and late term pregnant rats and the expression levels of GABA(A) alpha(1) and alpha(2) receptor subunits were analyzed using quantitative competitive reverse transcriptase polymerase chain reaction (RT-PCR) and immunoblot techniques. The competitive RT-PCR analysis allows comparison of expression levels between different mRNA, and the mRNA expression level of GABA(A) alpha(1) was several hundred fold greater than GABA(A) alpha(2) in both groups. However, this relative distribution of GABA(A) alpha(1) and alpha(2) receptor subunits protein or mRNA expression was not altered in late term pregnant compared to nonpregnant rats. These data demonstrate, that within the RVLM of both nonpregnant and late term pregnant rats, the relative expression levels of GABA(A) alpha(1,2) receptor subunits favor GABA(A) receptors susceptible to positive modulation by progesterone metabolites.
Foley
C Michael
CM
Department of Biomedical Sciences, Dalton Cardiovascular Research Center, 134 Research Park Drive, University of Missouri-Columbia, Columbia, MO 65211-3300, USA. foleycm@missouri.edu
Stanton
Jeffery J
JJ
Price
Elmer M
EM
Cunningham
J Thomas
JT
Hasser
Eileen M
EM
Heesch
Cheryl M
CM
D000818:Animals; D015153:Blotting, Western; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D005260:Female; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000096:biosynthesis / Q000235:genetics; D007091:Image Processing, Computer-Assisted; D008526:Medulla Oblongata / Q000378:metabolism*; D010625:Phenylethanolamine N-Methyltransferase / Q000096:biosynthesis; D011247:Pregnancy; D011270:Pregnancy, Animal / Q000378:metabolism*; D021122:Protein Subunits / Q000096:biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D011963:Receptors, GABA-A / Q000096:biosynthesis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014446:Tyrosine 3-Monooxygenase / Q000096:biosynthesis
D018076:DNA, Complementary; D021122:Protein Subunits; D012333:RNA, Messenger; D011963:Receptors, GABA-A; D014446:Tyrosine 3-Monooxygenase; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D010625:Phenylethanolamine N-Methyltransferase