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This Article Full Text Full Text (PDF) All Versions of this Article: 287/1/G178    most recent 00528.2003v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Web of Science (11) Citing Articles via Google Scholar Google Scholar Articles by De Jonge, F. Articles by Kroese, A. B. A. Search for Related Content PubMed PubMed Citation Articles by De Jonge, F. Articles by Kroese, A. B. A.

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In vitro activation of murine DRG neurons by CGRP-mediated mucosal mast cell degranulation

¹Laboratory of Cell Biology and Histology, Department of Biomedical Sciences, ²Laboratory of Electrobiology, University of Antwerp, B-2020 Antwerp, Belgium; ³Department of Veterinary Clinical Studies, Royal (Dick) School of Veterinary Studies, University of Edinburgh, Edinburgh EH25 9RG, United Kingdom; and ⁴Departments of Medical Physiology and Surgery, University Medical Centre, 3584 CG Utrecht, The Netherlands

Submitted 19 December 2003 ; accepted in final form 1 March 2004

Upregulation of CGRP-immunoreactive (IR) primary afferent nerve fibers accompanied by mastocytosis is characteristic for the Schistosoma mansoni-infected murine ileum. These mucosal mast cells (MMC) and CGRP-IR fibers, which originate from dorsal root (DRG) and nodose ganglia, are found in close apposition. We examined interactions between primary cultured MMC and CGRP-IR DRG neurons in vitro by confocal recording of intracellular Ca²⁺ concentration ([Ca²⁺]_i). The degranulatory EC₅₀ for the mast cell secretagogue compound 48/80 (C48/80; 10 µg/ml) and the neuropeptides CGRP (2.10^–8 M) and substance P (SP; 3.10^–8 M) were determined by measurement of extracellular release of the granule chymase, mouse mast cell protease-1. Application of C48/80 (10 µg/ml) and CGRP and SP (both 10^–7 M) to Fluo-4-loaded MMC induced a transient rise in [Ca²⁺]_i after a lag time, indicative of mast cell degranulation and/or secretion. The CGRP response could be completely blocked by pertussis toxin (2 µg/ml), indicating involvement of G_i proteins. Application of MMC juice, obtained by C48/80 degranulation of MMC, to Fluo-4-loaded DRG neurons induced in all neurons a rise in [Ca²⁺]_i, indicative of activation. Degranulation of MMC by C48/80 in culture dishes containing Fluo-4-loaded DRG neurons also caused activation of the DRG neurons. In conclusion, these results demonstrate a bidirectional cross-talk between cultured MMC and CGRP-IR DRG neurons in vitro. This indicates that such a communication may be the functional relevance for the close apposition between MMC and CGRP-IR nerve fibers in vivo.

calcium imaging; electrophysiology; cultured mucosal mast cells; cultured dorsal root ganglia neurons; mast cell juice

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