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The Journal of Neuroscience, August 1, 2001, 21(15):5461-5472

Developmental Regulation and Specific Brain Distribution of Phosphorabphilin

Davide L. Foletti and Richard H. Scheller

Howard Hughes Medical Institute, Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, California 94305-5428

Protein kinases and phosphatases play an important role in modulating synaptic transmission. The synaptic protein rabphilin associates with synaptic vesicles through the small GTPase Rab3A, binds Ca2+ and phospholipids, and interacts with cytoskeletal elements, yet its function remains controversial. In this study, we have generated phosphospecific antibodies and studied the developmental, subcellular, and brain distribution of rabphilin phosphorylated at serine-234 and serine-274. Our results show that phosphorabphilin is present in vivo under basal conditions in a specific subset of synapses. The phosphorylated rabphilin is abundant in the cerebellum, midbrain, and medulla; phosphorabphilin is specifically enriched in the climbing fiber synapses of the cerebellar cortex. Its developmental profile reveals a sharp and transient increase at approximately postnatal day 16, a period critical for the activity-dependent pruning of supernumerary climbing fibers in the cerebellum. We propose that the phosphorylation of rabphilin regulates neuronal activity through development and in a synapse-specific manner.

Key words: rabphilin; phosphospecific antibodies; protein kinases; development; climbing fibers; immunohistochemistry

Copyright © 2001 Society for Neuroscience  0270-6474/01/21155461-12$05.00/0


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