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蜜蜂培养的蕈形体肯扬细胞中的离子型GABA受体及其受细胞内钙的调节

An ionotropic GABA receptor in cultured mushroom body Kenyon cells of the honeybee and its modulation by intracellular calcium.

作者信息

Grünewald Bernd, Wersing Anna

机构信息

Institut für Biologie, Neurobiologie, Freie Universität Berlin, Königin-Luise-Str. 28-30, 14195, Berlin, Germany.

出版信息

J Comp Physiol A Neuroethol Sens Neural Behav Physiol. 2008 Apr;194(4):329-40. doi: 10.1007/s00359-007-0308-9. Epub 2008 Jan 5.

Abstract

GABAergic inhibitory transmission is very abundant within the insect brain. We, therefore, studied the functional properties of the ionotropic GABA receptor of honeybee mushroom body Kenyon cells in vitro. GABA applications elicit rapidly activating and desensitizing currents, which are concentration-dependent between 10 and 500 microM. The mean peak amplitude induced by 500 microM GABA at a holding potential of -110 mV is -1.55 +/- 0.23 nA (SEM, n = 29). The GABA-induced current is mediated by Cl(-) ions because (1) the reversal potential of the GABA-induced current of -40.6 mV is very close to the calculated Nernst potential of chloride (-44.8 mV). (2) With equimolar chloride concentrations the reversal potential shifted to about 0 mV. GABA or muscimol are equally efficient channel agonists, whereas CACA is a partial agonist. Picrotoxin or philanthotoxin (100 microM) completely and reversibly block the GABA-induced current, bicuculline (100 microM) has no effect. Elevating the intracellular Ca(2+) concentration increases the GABA current amplitude. This modulatory effect is blocked by the kinase blocker K 252a, but not by blockers of CaMkinaseII (KN-93), PKC (bisindolylmaleimide) or PKA (KT 5720). We conclude that Kenyon cells express functional GABA receptors whose properties support an inhibitory role of GABAergic transmission.

摘要

γ-氨基丁酸(GABA)能抑制性传递在昆虫大脑中非常丰富。因此,我们在体外研究了蜜蜂蘑菇体肯扬细胞的离子型GABA受体的功能特性。施加GABA会引发快速激活和脱敏电流,在10至500微摩尔之间呈浓度依赖性。在-110 mV的钳制电位下,500微摩尔GABA诱导的平均峰值幅度为-1.55±0.23纳安(标准误,n = 29)。GABA诱导的电流由Cl⁻离子介导,原因如下:(1)GABA诱导电流的反转电位为-40.6 mV,非常接近计算得出的氯离子能斯特电位(-44.8 mV)。(2)在等摩尔氯离子浓度下,反转电位移至约0 mV。GABA或蝇蕈醇是同等有效的通道激动剂,而CACA是部分激动剂。印防己毒素或 philanthotoxin(100微摩尔)能完全且可逆地阻断GABA诱导的电流,荷包牡丹碱(100微摩尔)则无作用。提高细胞内Ca²⁺浓度会增加GABA电流幅度。这种调节作用被激酶阻滞剂K 252a阻断,但未被CaM激酶II(KN-93)、蛋白激酶C(双吲哚马来酰亚胺)或蛋白激酶A(KT 5720)的阻滞剂阻断。我们得出结论,肯扬细胞表达功能性GABA受体,其特性支持GABA能传递的抑制作用。

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