Activation of P2-purino-, alpha 1-adreno and H1-histamine receptors triggers cytoplasmic calcium signalling in cerebellar Purkinje neurons.
The cytoplasmic calcium concentration ([Ca2+]i) was measured from Purkinje neurons in acutely prepared cerebellar slices. Neurons were loaded with calcium indicator Fura-2 by 40-min slice incubation in Tyrode solution containing 5 microM Fura-2/AM and 0.02% pluronic-F127. Bath applications of ATP (100 microM), epinephrine (10 microM) and histamine (100 microM) triggered a transient increase of [Ca2+]i in Purkinje neurons. ATP-induced [Ca2+]i elevation in Purkinje neurons was mimicked by ADP, but not AMP or adenosine pointing to the involvement of P2Y metabotropic purinoreceptors. Epinephrine-triggered [Ca2+]i responses were blocked by the selective alpha 1-antagonist prazosin and were mimicked by the alpha 1-adrenoreceptor agonist phenylephrine, and were not affected by beta- and alpha 2-adrenoreceptor agonists (isoproterenol and clonidine) and antagonists (propranolol and yohimbine). Histamine-induced [Ca2+]i responses demonstrated specific sensitivity to selective H1 antagonist chlorpheniramine, and were not sensitive to H2 and H3 histamine receptors modulators. The [Ca2+]i responses to all three agonists persisted in Ca(2+)-free extracellular media and were blocked by slice preincubation with thapsigargin (500 nM). We conclude that cerebellar Purkinje neurons are endowed with metabotropic P2 gamma purinoreceptors, alpha 1-adrenoreceptors and H1 histamine receptors which mediate the generation of intracellular [Ca2+]i signals via activation of Ca2+ release from inositol-1,4,5-trisphosphate-sensitive intracellular stores.