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Volume 16, Number 9, Issue of May 1, 1996 pp. 2891-2900
Copyright ©1996 Society for Neuroscience

Inositol 1,4,5-Trisphosphate-Gated Calcium Transport through Plasma Membranes in Nerve Terminals

Received Sept. 26, 1995; revised Feb. 5, 1996; accepted Feb. 7, 1996.

Hiroshi Ueda¹, Shigeki Tamura², Nobuyuki Fukushima¹, Toshiaki Katada³, Michio Ui⁴, and Masamichi Satoh⁵

¹ Department of Pharmacology, Yokohama City University School of Medicine, Yokohama 236, ² Suntory Institute for Biomedical Research, Osaka 618, ³ Department of Physiological Chemistry, Faculty of Pharmaceutical Sciences, The University of Tokyo, Tokyo 113, ⁴ RIKEN, Wako 351-01, and ⁵ Department of Molecular Pharmacology, Faculty of Pharmaceutical Sciences, Kyoto University, Kyoto 606-01, Japan

We developed new biochemical approaches to demonstrate the presence of inositol 1,4,5-trisphosphate (InsP₃)-gated calcium channels in presynaptic plasma membranes (SPM) and their involvement in the presynaptic receptor-mediated Ca²⁺ influx into nerve terminals. In perfusion experiments using SPM vesicles preloaded with ⁴⁵Ca²⁺, InsP₃ elicited the release of ⁴⁵Ca²⁺ into perfusates in a saturable manner. The InsP₃-evoked ⁴⁵Ca²⁺ release from resealed SPM vesicles was more potent than that from resealed vesicles using any other subcellular fractions. Here we also report the involvement of InsP₃-gated mechanisms in the presynaptic receptor-mediated Ca²⁺ influx into synaptosomes (nerve terminals) by use of such resealed vesicles reconstituted with purified G_i1.

Key words: InsP₃ receptor; presynaptic receptor; G_i1; reconstitution; resealed vesicles; Ca²⁺

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