CGRP1 receptor activation induces piecemeal release of protease-1 from mouse bone marrow-derived mucosal mast cells.
BACKGROUND
The parasitized or inflamed gastrointestinal mucosa shows an increase in the number of mucosal mast cells (MMC) and the density of extrinsic primary afferent nerve fibers containing the neuropeptide, calcitonin gene-related peptide (CGRP). Currently, the mode of action of CGRP on MMC is unknown.


METHODS
The effects of CGRP on mouse bone marrow-derived mucosal mast cells (BMMC) were investigated by measurements of intracellular Ca(2+)[Ca(2+)](i) and release of mMCP-1.


RESULTS
Bone marrow-derived mucosal mast cells responded to the application of CGRP with a single transient rise in [Ca(2+)](i). The proportion of responding cells increased concentration-dependently to a maximum of 19 ± 4% at 10(-5)mol L(-1) (mean ±SEM; C48/80 100%; EC(50)10(-8) mol L(-1) ). Preincubation with the CGRP receptor antagonist BIBN4096BS (10(-5) mol L(-1)) completely inhibited BMMC activation by CGRP [range 10(-5) to 10(-11) mol L(-1); analysis of variance (ANOVA) P < 0.001], while preincubation with LaCl(3) to block Ca(2+) entry did not affect the response (P = 0.18). The presence of the CGRP1 receptor on BMMC was confirmed by simultaneous immunofluorescent detection of RAMP1 or CRLR, the two components of the CGRP1 receptor, and mMCP-1. Application of CGRP for 1 h evoked a concentration-dependent release of mMCP-1 (at EC(50) 10% of content) but not of β-hexosaminidase and alterations in granular density indicative of piecemeal release.


CONCLUSIONS
We demonstrate that BMMC express functional CGRP1 receptors and that their activation causes mobilization of Ca(2+) from intracellular stores and piecemeal release of mMCP-1. These findings support the hypothesis that the CGRP signaling from afferent nerves to MMC in the gastrointestinal wall is receptor-mediated.
Rychter
J W
JW
Institute for Risk Assessment Sciences, Utrecht University, Utrecht, The Netherlands.
Van Nassauw
L
L
Timmermans
J-P
JP
Akkermans
L M A
LM
Westerink
R H S
RH
Kroese
A B A
AB
D000818:Animals; D001853:Bone Marrow / Q000378:metabolism*; D000077221:Calcitonin Gene-Related Peptide Receptor Antagonists; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D053818:Chymases / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D008297:Male; D008407:Mast Cells / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D010879:Piperazines / Q000494:pharmacology; D011799:Quinazolines / Q000494:pharmacology; D018015:Receptors, Calcitonin Gene-Related Peptide / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology; D001619:beta-N-Acetylhexosaminidases / Q000378:metabolism
D000077221:Calcitonin Gene-Related Peptide Receptor Antagonists; D010879:Piperazines; D011799:Quinazolines; D018015:Receptors, Calcitonin Gene-Related Peptide; D001619:beta-N-Acetylhexosaminidases; D053818:Chymases; C509230:Mcpt1 protein, mouse; D002118:Calcium; C406305:olcegepant