Calcitonin gene-related peptide receptor in human placental syncytiotrophoblast brush-border and basal plasma membranes.
Minerals, such as calcium and potassium, are essential for fetal development, but their transplacental transport, and in particular, the effect of hormones on this process has not been extensively studied. Human alpha-calcitonin gene-related peptide (h alpha CGRP), a hormone constituted of 37 amino acids, is obtained by the alternative splicing of the mRNA from the calcitonin gene, and could be implicated in placental ion transport. In order to study the presence of this receptor, brush-border and basal plasma membranes were purified, and membrane binding studies were conducted using [125I]h alpha CGRP. The initiation of binding of [125I]h alpha CGRP to both membranes was rapid and reached maximal value after 10 min of incubation at 37 degrees C. Scratchard analysis revealed single-affinity binding sites for h alpha CGRP with Kd equal to 4412.45 +/- 604.81 pM and 2673.24 +/- 552.51 pM for brush-border and basal plasma membranes, respectively, which were significantly different. Moreover, the maximal number of receptors was significantly different (P < 0.001) in both membranes, with Bmax of 627.94 +/- 31.40 fmol/mg protein for brush-border membranes and 343.70 +/- 43.52 fmol/mg protein in basal-plasma membranes. Competitive displacement of [125I]h alpha CGRP with other ligands showed the following potencies; h alpha CGRP approximately h beta CGRP approximately Cys (acm)2,7 CGRP > CGRP (8-37), but no competition was observed with human and salmon calcitonin. Half-maximal displacement for human alpha- and beta CGRP was reached at approximately 10(-10)M for brush-border and basal-plasma membranes. alpha- and beta CGRP, and their fragment and analogue, stimulated cyclic AMP production in placental homogenate ranging from 143-163 per cent. Thus, our results show the presence of CGRP-specific receptors in both the syncytiotrophoblast membranes of human placenta. The role(s) of this related peptide in placenta remains to be investigated.
Lafond
J
J
Department of Biological Sciences, University of Quebec in Montreal.
St-Pierre
S
S
Masse
A
A
Savard
R
R
Simoneau
L
L
D001667:Binding, Competitive; D015740:Calcitonin Gene-Related Peptide / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000096:biosynthesis; D005260:Female; D006801:Humans; D008024:Ligands; D008871:Microvilli / Q000378:metabolism; D011247:Pregnancy; D018015:Receptors, Calcitonin Gene-Related Peptide / Q000378:metabolism*; D014327:Trophoblasts / Q000378:metabolism* / Q000648:ultrastructure
D008024:Ligands; D018015:Receptors, Calcitonin Gene-Related Peptide; D000242:Cyclic AMP; D015740:Calcitonin Gene-Related Peptide