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J Neurophysiol 86: 1389-1397, 2001;
0022-3077/01 $5.00
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The Journal of Neurophysiology Vol. 86 No. 3 September 2001, pp. 1389-1397
Copyright ©2001 by the American Physiological Society

Characterization of the mGluR1-Mediated Electrical and Calcium Signaling in Purkinje Cells of Mouse Cerebellar Slices

F. Tempia,1,2 M. E. Alojado,1 P. Strata,2 and T. Knöpfel1

 1Laboratory for Neuronal Circuit Dynamics, Brain Science Institute, RIKEN, Saitama 351-0198, Japan; and  2Department of Neuroscience and Rita Montalcini Centre for Brain Repair, University of Turin, I-10125 Turin, Italy

Tempia, F., M. E. Alojado, P. Strata, and T. Knöpfel. Characterization of the mGluR1-Mediated Electrical and Calcium Signaling in Purkinje Cells of Mouse Cerebellar Slices. J. Neurophysiol. 86: 1389-1397, 2001. The metabotropic glutamate receptor 1 (mGluR1) plays a fundamental role in postnatal development and plasticity of ionotropic glutamate receptor-mediated synaptic excitation of cerebellar Purkinje cells. Synaptic activation of mGluR1 by brief tetanic stimulation of parallel fibers evokes a slow excitatory postsynaptic current and an elevation of intracellular calcium concentration ([Ca2+]i) in Purkinje cells. The mechanism underlying these responses has not been identified yet. Here we investigated the responses to synaptic and direct activation of mGluR1 using whole cell patch-clamp recordings in combination with microfluorometric measurements of [Ca2+]i in mouse Purkinje cells. Following pharmacological block of ionotropic glutamate receptors, two to six stimuli applied to parallel fibers at 100 Hz evoked a slow inward current that was associated with an elevation of [Ca2+]i. Both the inward current and the rise in [Ca2+]i increased in size with increasing number of pulses albeit with no clear difference between the minimal number of pulses required to evoke these responses. Application of the mGluR1 agonist (S)-3,5-dihydroxyphenylglycine (3,5-DHPG) by means of short-lasting (5-100 ms) pressure pulses delivered through an agonist-containing pipette positioned over the Purkinje cell dendrite, evoked responses resembling the synaptically induced inward current and elevation of [Ca2+]i. No increase in [Ca2+]i was observed with inward currents of comparable amplitudes induced by the ionotropic glutamate receptor agonist AMPA. The 3,5-DHPG-induced inward current but not the associated increase in [Ca2+]i was depressed when extracellular Na+ was replaced by choline, but, surprisingly, both responses were also depressed when bathing the tissue in a low calcium (0.125 mM) or calcium-free/EGTA solution. Thapsigargin (10 µM) and cyclopiazonic acid (30 µM), inhibitors of sarco-endoplasmic reticulum Ca2+-ATPase, had little effect on either the inward current or the elevation in [Ca2+]i induced by 3,5-DHPG. Furthermore, the inward current induced by 3,5-DHPG was neither blocked by 1-[2-(4-methoxyphenyl)-2-[3-(4-methoxyphenyl)propoxy] ethyl-1H-imidazole, an inhibitor of store operated calcium influx, nor by nimodipine or omega-agatoxin, blockers of voltage-gated calcium channels. These electrophysiological and Ca2+-imaging experiments suggest that the mGluR1-mediated inward current, although mainly carried by Na+, involves influx of Ca2+ from the extracellular space.




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