6533b7d0fe1ef96bd125b837

RESEARCH PRODUCT

Changes in cytosolic calcium in response to noxious heat and their relationship to vanilloid receptors in rat dorsal root ganglion neurons.

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Heat transduction mechanisms in primary nociceptive afferents have been suggested to involve a vanilloid receptor channel with high calcium permeability. To characterize the changes in free cytosolic calcium evoked by noxious heat stimuli (< or =51 degrees C, 10s), we performed microfluorometric measurements in acutely dissociated small dorsal root ganglion neurons (< or =32.5 microm) of adult rats using the dye FURA-2. Only neurons that responded with a reversible increase in intracellular calcium to high potassium were evaluated. Heat-induced calcium transients (exceeding mean + 3S.D. of the temperature dependence of the dye) were found in 66 of 105 neurons. These transients increased non-linearly with temperature. In contrast, heat-insensitive neurons showed a small linear increase of intracellular calcium throughout the range of 12-49 degrees C, similar to cardiac muscle cells. The vanilloid receptor agonist capsaicin induced calcium transients in 72 of 99 neurons. Capsaicin sensitivity and heat sensitivity were significantly associated (P<0.001, chi(2)-test), but 16 of 34 heat-insensitive cells responded to capsaicin and four of 49 heat-sensitive cells were capsaicin insensitive. The competitive vanilloid receptor antagonist capsazepine (10 microM) reversibly reduced the heat-induced calcium transients by 47+/-13%. In contrast, high potassium-induced calcium transients were not affected by pre-incubation with capsazepine. In calcium-free extracellular solution, the heat-induced rise in intracellular calcium was reduced by 76+/-5%. Heat-induced calcium transients were also reversibly reduced by 75+/-6% in sodium-free solution and by 62+/-7% with the L-type calcium channel blocker nifedipine (5 microM). These results indicate that noxious heat rapidly increases intracellular calcium in nociceptive primary sensory neurons. Heat-sensitive vanilloid receptors are involved in the induction of calcium transients, and calcium is also released from intracellular stores, but the main fraction of calcium passes through voltage-operated calcium channels.