Identification of Neurotransmitter released from hair cells

鉴定毛细胞释放的神经递质

• 批准号: 06454713
• 负责人: OHMORI Harunori
• 金额: $ 1.34万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06454713/
• 关键词: Hair cell; cerebellum; granule cells; Purkinje cells; glutamate; NMDA; 小脳顆粒細胞; 神経伝達物質

In order to determine the neurotransmitter released from auditory hair cells, we have invented an artificial synapse formed between isolated hair cells and cultured cerebellar granule cells. Both hair cells and granule cells were whole cell voltage clamped, and the current detected in the granule cells in response to the hair cell membrane depolarization were analyzed. Outward current was detected in the granule cells voltaged clamped at positive membrane potentials. This current was blocked reversibly by the bath applied DL-APV,therefore, concluded as due to the activation of NMDA type glutamate receptors on the granule cell. From detailed investigations of the dose-dependencies of several excitatory amino acids for NMDA and non-NMDA receptors expressed on the granule cells and on the Purkinje cells, respectively, glutamate was concluded as the most likely neurotransmitter released from hair cells. When glutamate was released from hair cells, it could mobilized up to 860 synaptic vesicles in 0.5 sec and such vesicles are likely fused with the presynaptic membrane at a rate of 500 Hz at each active zones in the hair cell.

为了测定听毛细胞释放的神经递质，我们发明了一种在分离的毛细胞和培养的小脑颗粒细胞之间形成的人工突触。对毛细胞和颗粒细胞进行全细胞电压钳位，并分析颗粒细胞中检测到的响应于毛细胞膜去极化的电流。颗粒细胞在正膜电位电压钳制下可检测到外向电流。该电流被浴中施加的DL-APV可逆地阻断，因此，得出结论，这是由于颗粒细胞上的NMDA型谷氨酸受体的激活。从几种兴奋性氨基酸的剂量依赖性的详细调查的颗粒细胞和浦肯野细胞上表达的NMDA和非NMDA受体，分别，谷氨酸的结论是最有可能的神经递质从毛细胞释放。当谷氨酸从毛细胞释放时，它可以在0.5秒内动员多达860个突触小泡，并且这些小泡可能以500 Hz的速率在毛细胞的每个活性区与突触前膜融合。

项目成果

Kataoka, Y. & Ohmori, H.: "Activation of glutamate reugtors in response to membrane depolaricution of hair cells isolated from chick cochlea." Journal of Physiology. 447. 403-414 (1994)

片冈，Y.

Kataoka, Y.& Ohmori, H.: "Activation of glutamate receptors in response to membrane depolarization of hair cells isolated from chick cochlea." Journal of Physiology. 477. 403-414 (1994)

片冈，Y.

Ohmori, H.: "Active Hearing. eds. & Flock, D. DTToson, M. Ulfendahl" Pergamion, 376 (1995)

Ohmori, H.：“主动听力。编辑。

Yoshida, N., Shigemoto T. Sugai, T. & Ohmori, N.: "The role of Tnositol trisphosphate on Ach-induced outward current in bull frog saccular hair cells" Brain Research. 644. 90-100 (1994)

吉田，N.，茂本 T. 菅井，T.

大森治紀: "内耳有毛細胞におけるシナプス伝達機構の研究" 日本生理学雑誌. 56. 197-205 (1995)

Harunori Omori：“内耳毛细胞突触传递机制的研究”日本生理学杂志 56. 197-205 (1995)。