Expression of adrenomedullin (ADM) and its binding sites in the rat uterus: increased number of binding sites and ADM messenger ribonucleic acid in 20-day pregnant rats compared with nonpregnant rats.
RIA of nonpregnant rat uterus extracts showed 0.68 +/- 0.08 pmol/g adrenomedullin (ADM) and 3.23 +/- 0.08 pmol/g calcitonin gene-related peptide (CGRP). In the pregnant (20 days gestation) uterus, the ADM content was 0.90 +/- 0.17 pmol/g, and CGRP could not be detected. ADM messenger RNA was detected at high levels in the uterus, with a 1.8-fold increase in expression in pregnancy. Pharmacologically distinct binding sites for ADM (Bmax = 21 +/- 2 fmol/mg protein, dissociation constant = 80 +/- 6 pM), and CGRP (Bmax = 101 +/- 18 fmol/mg protein, dissociation constant = 140 +/- 20 pM) were identified in nonpregnant uterus. Competition for 125I[Tyr0]alphaCGRP binding was shown by both ADM and CGRP (8-37), whereas CGRP and CGRP (8-37) did not compete for 125I-ADM-binding sites. The density of the ADM-binding sites was 10 times greater in pregnant uterus (Bmax = 211 +/- 39 fmol/mg protein, P < 0.01) than nonpregnant uterus. CGRP receptor messenger RNA was identified in both nonpregnant and pregnant uteri. In isolated nonpregnant rat uteri, CGRP and ADM attenuated the contractile response to galanin by 77 +/- 10% and 57 +/- 10%, respectively. The responses to both CGRP and ADM were abolished by CGRP (8-37). These results demonstrate, for the first time, the presence of ADM and specific binding sites for both ADM and CGRP in the rat uterus.
Upton
P D
PD
Division of Endocrinology and Metabolic Medicine, Royal Postgraduate Medical School, Hammersmith Hospital, London, United Kingdom.
Austin
C
C
Taylor
G M
GM
Nandha
K A
KA
Clark
A J
AJ
Ghatei
M A
MA
Bloom
S R
SR
Smith
D M
DM
D053607:Adrenomedullin; D000818:Animals; D015152:Blotting, Northern; D015740:Calcitonin Gene-Related Peptide / Q000096:biosynthesis / Q000494:pharmacology; D005260:Female; D019004:Galanin / Q000494:pharmacology; D006801:Humans; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000378:metabolism; D010452:Peptide Biosynthesis; D010455:Peptides / Q000378:metabolism / Q000494:pharmacology; D010920:Placenta / Q000378:metabolism*; D011247:Pregnancy; D011270:Pregnancy, Animal / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis; D011863:Radioimmunoassay; D051381:Rats; D017208:Rats, Wistar; D058265:Receptors, Adrenomedullin; D018000:Receptors, Peptide; D012016:Reference Values; D014158:Transcription, Genetic / Q000187:drug effects; D014590:Uterine Contraction / Q000187:drug effects; D014599:Uterus / Q000187:drug effects / Q000378:metabolism*
D008565:Membrane Proteins; D010455:Peptides; D012333:RNA, Messenger; D058265:Receptors, Adrenomedullin; D018000:Receptors, Peptide; D053607:Adrenomedullin; D019004:Galanin; D015740:Calcitonin Gene-Related Peptide