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J Biol Chem, Vol. 273, Issue 42, 27620-27624, October 16, 1998

Brain-derived Neurotrophic Factor Induces Rapid and Transient Release of Glutamate through the Non-exocytotic Pathway from Cortical Neurons

Nobuyuki Takei^¶, Tadahiro Numakawa^¶, Shunji Kozaki, Naoto Sakai^¶, Yasuhisa Endo, Masami Takahashi^**, and Hiroshi Hatanaka^¶

From the  Department of Applied Biology, Kyoto Institute of Technology, Sakyo, Kyoto 606, ^¶ Division of Protein Biosynthesis, Institute for Protein Research, Osaka University, Suita, Osaka 565,  Department of Veterinary Science, College of Agriculture, University of Osaka Prefecture, Sakai, Osaka 593, and ^** Mitsubishi Kasei Institute for Life Sciences, Machida, Tokyo 194, Japan

There is increasing interest in the involvement of neurotrophins in neural transmission and plasticity. Thus, we investigated the effects of brain-derived neurotrophic factor (BDNF) on glutamate release from cortical neurons. Treatment of cultured cortical neurons with BDNF induced rapid and transient release of glutamate. This effect was suggested to be mediated by TrkB activation because K252a inhibited the release of glutamate and BDNF phosphorylated TrkB within 30 s. BDNF-induced glutamate release was observed even when using Ca²⁺-free assay buffer but was inhibited by BAPTA-AM, a cell-permeable Ca²⁺ chelator. Therefore, BDNF-induced glutamate release was independent of extracelluar Ca²⁺ but dependent on intracellular Ca²⁺. Because normal neurotransmitter release is exocytotic, the involvement of the exocytotic pathway in BDNF-induced glutamate release was examined. As botulinum toxin is known to cleave exocytosis-associated proteins, thereby inhibiting exocytosis, it was applied to neurons prior to the release assay. Although botulinum toxin B cleaved VAMP2 and inhibited Ca²⁺-triggered glutamate release, it did not inhibit the BDNF-induced release of glutamate. These results strongly suggested that BDNF induces rapid and transient release of glutamate from cortical neurons through a non-exocytotic pathway.

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