Dendritic cell development: multiple pathways to nature's adjuvants

The endothelin (ET) isoforms ET-1, ET-2 and ET-3 applied at 100 nM triggered a transient increase in Ca2+]i in Bergmann glial cells in cerebellar slices acutely isolated from 20-25 day-old mice. The intracellular calcium concentration (Ca2+]i) was monitored using Fura-2-based Ca2+]i microfluorimetry. The ET-triggered Ca2+]i transients were mimicked by ETB receptor agonist BQ-3020 and were inhibited by ETB receptor antagonist BQ-788. ET elevated Ca2+]i in Ca(2+)-free extracellular solution and the ET-triggered Ca2+]i elevation was blocked by 500 nM thapsigargin indicating that the Ca2+]i was released from InsP3-sensitive intracellular pools. The ET-triggered Ca2+]i increase in Ca(2+)-free solution was shorter in duration. Restoration of normal extracellular Ca2+] briefly after the ET application induced a second Ca2+]i increase indicating the presence of a secondary Ca2+ influx which prolongs the Ca2+ signal. Pre-application of 100 microM ATP or 10 microM noradrenaline blocked the ET response suggesting the involvement of a common Ca2+ depot. The expression of ETB receptor mRNAs in Bergmann glial cells was revealed by single-cell RT-PCR. The mRNA was also found in Purkinje neurones, but no Ca2+ signalling was triggered by ET. We conclude that Bergmann glial cells are endowed with functional ETB receptors which induce the generation of intracellular Ca2+]i signals by activation of Ca2+ release from InsP3-sensitive intracellular stores followed by a secondary Ca2+ influx.