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Articles in PresS, published online ahead of print August 14, 2002
Am J Physiol Cell Physiol, 10.1152/ajpcell.00050.2002
Submitted on January 31, 2002
Accepted on July 17, 2002
1 Analytical Chemistry & Biophysics, Nagoya City University, Nagoya, Japan
2 Intestinal Diseases Research Programme, McMaster University, Hamilton, ON, Canada
3 National Institute of Health Sciences, Tokyo, Japan
* To whom correspondence should be addressed. E-mail: bienens{at}mcmaster.ca.
Mast cell-neurite interaction has served as a model for neuro-immune interaction. We showed that neurite-mast cell communication can occur via substance P interacting with neurokinin (NK)-1 receptors on the mucosal mast cell-like cell, the rat basophilic leukemia (RBL) cell. Here neurite (murine superior cervical ganglia) and RBL cell (expressing the granule-associated antigen CD63-green fluorescent protein (GFP) conjugate) co-cultures were established and stimulated with bradykinin (BK, 10 nM) or scorpion venom (SV; 10 pg/ml), both of which only activate neurites. Cell activation was assessed by confocal imaging of Ca2+ (cells preloaded with Fluo-3) and analyses of RBL CD63-GFP+ granule movement conducted. Neurite activation by BK or SV was followed by RBL Ca2+ mobilization and this was inhibited by an NK-1 receptor antagonist (NK-1 RA). Moreover, membrane ruffling was observed on RBL pseudopodial extensions in contact with the activated neurite, but not on non-contacting pseudopodia. RBL membrane ruffling was inhibited by the NK-1 RA, but not an NK-2 RA, and was accompanied by a significant increase in granule movement (0.13 ± 0.04* vs. 0.05 ± 0.01 µm/sec) that was most evident at the point of neurite contact: many of the granules moved towards the plasmalemma. This is the first documentation of such precise (restricted to the membranes contact site) transfer of information between nerves and mast cells that could allow for very subtle in vivo communication between these two cell types.
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