Gestational changes in uterine L-type calcium channel function and expression in guinea pig.
Pregnancy can influence both the resting membrane potential and the ion channel composition of the uterine myometrium. Calcium flux is essential for excitation-contraction coupling in pregnant uterus. The uterine L-type calcium channel is an important component in mediating calcium flux and is purported to play a role in parturition. This study was undertaken to characterize gestational changes in 1) the uterine contractile response to the L-type calcium channel agonist, Bay K 8644; 2) the mRNA expression of channel subunits by semiquantitative reverse transcriptase-polymerase chain reaction; and 3) estimate channel protein levels by measuring (3)H-isradipine binding at the dihydropyridine binding site of the alpha(1c) subunit utilizing saturation binding methods. Sensitivity to Bay K 8644 increases beginning at 0.8 of gestation and persists through term. The change in sensitivity is coincident with an increased mRNA expression of the alpha(1c) and beta(2) subunits but with the least detectable amounts of isradipine binding. The expressed alpha(1c) transcript represents a novel structural variant with a 118-amino acid deletion in the III-IV linker and repeats IVS1-S3 of the protein sequence. The guinea pig uterine L-type calcium channel activity is highly regulated through gestation, but the regulation of mRNA expression may be different from regulation of protein levels, estimated by isradipine binding. The up-regulation of function, alpha(1c) subunit mRNA expression, and isradipine binding at term gestation are consistent with a role for this ion channel in parturition.
Collins
P L
PL
Departments of Obstetrics/Gynecology, Pediatrics, Biochemistry, and the Rammelcamp Center for Research, MetroHealth Campus, Case Western Reserve University, Cleveland, Ohio 44109, USA. pcollin@luc.edu
Moore
J J
JJ
Lundgren
D W
DW
Choobineh
E
E
Chang
S M
SM
Chang
A S
AS
D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester / Q000378:metabolism / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D002120:Calcium Channel Agonists / Q000494:pharmacology; D002121:Calcium Channel Blockers / Q000378:metabolism; D020746:Calcium Channels, L-Type / Q000096:biosynthesis / Q000378:metabolism*; D004095:Dihydropyridines / Q000378:metabolism; D005260:Female; D006168:Guinea Pigs; D017275:Isradipine / Q000378:metabolism; D008969:Molecular Sequence Data; D011247:Pregnancy; D011270:Pregnancy, Animal / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis; D011817:Rabbits; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014590:Uterine Contraction / Q000187:drug effects; D014599:Uterus / Q000378:metabolism*
D002120:Calcium Channel Agonists; D002121:Calcium Channel Blockers; D020746:Calcium Channels, L-Type; D004095:Dihydropyridines; D012333:RNA, Messenger; D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester; D017275:Isradipine