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Dendritic differentiation of cerebellar Purkinje cells is promoted by ryanodine receptors expressed by Purkinje and granule cells
Authors:Ryo Ohashi  Shin‐ichi Sakata  Asami Naito  Naohide Hirashima  Masahiko Tanaka
Affiliation:Department of Cellular Biophysics, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya, Japan
Abstract:Cerebellar Purkinje cells have the most elaborate dendritic trees among neurons in the brain. We examined the roles of ryanodine receptor (RyR), an intracellular Ca2+ release channel, in the dendrite formation of Purkinje cells using cerebellar cell cultures. In the cerebellum, Purkinje cells express RyR1 and RyR2, whereas granule cells express RyR2. When ryanodine (10 µM), a blocker of RyR, was added to the culture medium, the elongation and branching of Purkinje cell dendrites were markedly inhibited. When we transferred small interfering RNA (siRNA) against RyR1 into Purkinje cells using single‐cell electroporation, dendritic branching but not elongation of the electroporated Purkinje cells was inhibited. On the other hand, transfection of RyR2 siRNA into granule cells also inhibited dendritic branching of Purkinje cells. Furthermore, ryanodine reduced the levels of brain‐derived neurotrophic factor (BDNF) in the culture medium. The ryanodine‐induced inhibition of dendritic differentiation was partially rescued when BDNF was exogenously added to the culture medium in addition to ryanodine. Overall, these results suggest that RyRs expressed by both Purkinje and granule cells play important roles in promoting the dendritic differentiation of Purkinje cells and that RyR2 expressed by granule cells is involved in the secretion of BDNF from granule cells. © 2013 Wiley Periodicals, Inc. Develop Neurobiol 74: 467–480, 2014
Keywords:ryanodine receptor  dendrite  Purkinje cell  granule cell  single‐cell electroporation
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