Nerve-mast cell (RBL) interaction: RBL membrane ruffling occurs at the contact site with an activated neurite

Abstract

Mast cell-neurite interaction serves as a model for neuroimmune interaction. We have shown 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. Neurite (murine superior cervical ganglia) and RBL cell [expressing the granule-associated antigen CD63-green fluorescent protein (GFP) conjugate] cocultures were established and stimulated with bradykinin (BK; 10 nM) or scorpion venom (SV; 10 pg/ml), both of which activate only neurites. Cell activation was assessed by confocal imaging of Ca 2ϩ (cells preloaded with fluo 3), and analyses of RBL CD63-GFP ϩ granule movement were conducted. Neurite activation by BK or SV was followed by RBL Ca 2ϩ mobilization, which 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 noncontacting pseudopodia. RBL membrane ruffling was inhibited by NK-1 RA, but not NK-2 RA, and was accompanied by a significant increase in granule movement (0.13 Ϯ 0.04 vs. 0.05 Ϯ 0.01 m/s) that was most evident at the point of neurite contact: many of the granules moved toward the plasmalemma. This is the first documentation of such precise (restricted to the membrane's contact site) transfer of information between nerves and mast cells that could allow for very subtle in vivo communication between these two cell types. neuroimmunity; substance P; neurokinin-1 receptor; CD63; granule tracking ANALYSES OF NERVE-MAST CELL INTERACTIONS have been of significant importance in unequivocally establishing the concept of bidirectional neuroimmune communication (13,25). In addition to the anatomic association of mast cells and nerve fibers in many tissues (3,20), numerous studies have shown that mast cell activation can be evoked by nerve stimulation or the application of neurotransmitters and that mast cell-derived mediators can influence neuronal activity (7,18,24). For instance, the neuropeptide substance P has been shown to cause mast cell degranulation when used at high doses, whereas exposure to picomolar concentrations of substance P primes the mast cell, lowering the degranulation stimulation threshold to a second stimulus (10). We have used an in vitro model of mast cell-nerve interaction, composed of cocultures of the mucosal mast cell-like rat basophilic leukemic (RBL) cells and neurite-sprouting murine superior cervical ganglia (5). We showed that nerve-mast cell communication did not require transduction by an intermediate cell and that the RBL cell activation response following neurite stimulation was mediated largely via substance P release and through neurokinin (NK)-1 receptors (21). Here, we sought to further examine neurite-RBL cell interactions, to determine whether neuroninduced mast cell activation is a generalized whole cell response or if there is an additional level of subtlety to the neurite-RBL cell interaction that occurs at the specific...

Mast Cell-Dependent Excitation of Visceral-Nociceptive Sensory Neurons in Irritable Bowel Syndrome

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Substance P can stimulate prostaglandin D2 and leukotriene C4 generation without granule exocytosis in murine mast cells

“…The neural network and mast cells form an interactive unit that is able to influence immune and inflammatory responses (Mori et al, 2002). Neuropeptides released from sensory neurons can induce mast cell activation and their mediators can further stimulate the release of neuropeptides from sensory nerves resulting in a bi-directional mast cell -sensory neuron autocatalytic loop (Ansel et al, 1993(Ansel et al, , 1996Johnson and Erdos, 1973;Kowalski and Kaliner, 1988).…”

N-Cadherin Plays a Role in the Synapse-Like Structures between Mast Cells and Neurites

“…[6][7][8] In addition we studied by atomic force microscopy (AFM) the morphological structure of the interaction between neurites and mast cells (RBL) which was involved in the communication.…”