The Journal of General Physiology
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Published 1 March 2000. doi:10.1085/jgp.115.3.339
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© The Rockefeller University Press, 0022-1295/2000//339/ $5.00
Journal of General Physiology, Volume 115, Number 3, 2000

Original Article

Impaired Calcium Release in Cerebellar Purkinje Neurons Maintained in Culture

Mary D. Womacka, Jeffery W. Walkerb, and Kamran Khodakhaha

a Department of Physiology & Biophysics, University of Colorado Health Sciences Center, Denver, Colorado 80262
b Department of Physiology, University of Wisconsin, Madison, Wisconsin 53706
Department of Physiology & Biophysics, University of Colorado Health Sciences Center, 4200 East 9th Ave., C-240, Denver, CO 80262.303-315-8110

kamran.khodakhah{at}uchsc.edu

Cerebellar Purkinje neurons demonstrate a form of synaptic plasticity that, in acutely prepared brain slices, has been shown to require calcium release from the intracellular calcium stores through inositol trisphosphate (InsP3) receptors. Similar studies performed in cultured Purkinje cells, however, find little evidence for the involvement of InsP3 receptors. To address this discrepancy, the properties of InsP3- and caffeine-evoked calcium release in cultured Purkinje cells were directly examined. Photorelease of InsP3 (up to 100 µM) from its photolabile caged analogue produced no change in calcium levels in 70% of cultured Purkinje cells. In the few cells where a calcium increase was detected, the response was very small and slow to peak. In contrast, the same concentration of InsP3 resulted in large and rapidly rising calcium responses in all acutely dissociated Purkinje cells tested. Similar to InsP3, caffeine also had little effect on calcium levels in cultured Purkinje cells, yet evoked large calcium transients in all acutely dissociated Purkinje cells tested. The results demonstrate that calcium release from intracellular calcium stores is severely impaired in Purkinje cells when they are maintained in culture. Our findings suggest that cultured Purkinje cells are an unfaithful experimental model for the study of the role of calcium release in the induction of cerebellar long term depression.

Key Words: inositol trisphosphate • ryanodine • long term depression • synaptic plasticity • calcium stores

© 2000 The Rockefeller University Press


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