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大鼠海马中ATP敏感性钾通道的细胞类型特异性表达。

Cell-type specific expression of ATP-sensitive potassium channels in the rat hippocampus.

作者信息

Zawar C, Plant T D, Schirra C, Konnerth A, Neumcke B

机构信息

I. Physiologisches Institut der Universitat des Saarlandes, D-66421 Homburg, Germany.

出版信息

J Physiol. 1999 Jan 15;514 ( Pt 2)(Pt 2):327-41. doi: 10.1111/j.1469-7793.1999.315ae.x.

Abstract
  1. The distribution of ATP-sensitive K+ channels (KATP channels) was investigated in four cell types in hippocampal slices prepared from 10- to 13-day-old rats: CA1 pyramidal cells, interneurones of stratum radiatum in CA1, complex glial cells of the same area and granule cells of the dentate gyrus. The neuronal cell types were identified visually and characterized by the shapes and patterns of their action potentials and by neurobiotin labelling. 2. The patch-clamp technique was used to study the sensitivity of whole-cell currents to diazoxide (0.3 mM), a KATP channel opener, and to tolbutamide (0.5 mM) or glibenclamide (20 microM), two KATP channel inhibitors. The fraction of cells in which whole-cell currents were activated by diazoxide and inhibited by tolbutamide was 26% of pyramidal cells, 89 % of interneurones, 100% of glial cells and 89% of granule cells. The reversal potential of the diazoxide-induced current was at the K+ equilibrium potential and a similar current activated spontaneously when cells were dialysed with an ATP-free pipette solution. 3. Using the single-cell RT-PCR method, the presence of mRNA encoding KATP channel subunits (Kir6.1, Kir6.2, SUR1 and SUR2) was examined in CA1 pyramidal cells and interneurones. Subunit mRNA combinations that can result in functional KATP channels (Kir6.1 together with SUR1, Kir6.2 together with SUR1 or SUR2) were detected in only 17% of the pyramidal cells. On the other hand, KATP channels may be formed in 75% of the interneurones, mainly by the combination of Kir6.2 with SUR1 (58% of all interneurones). 4. The results of these combined analyses indicate that functional KATP channels are present in principal neurones, interneurones and glial cells of the rat hippocampus, but at highly different densities in the four cell types studied.
摘要
  1. 研究了从10至13日龄大鼠制备的海马切片中四种细胞类型的ATP敏感性钾通道(KATP通道)分布:CA1锥体细胞、CA1放射层中间神经元、同一区域的复合胶质细胞和齿状回颗粒细胞。通过视觉识别神经元细胞类型,并根据其动作电位的形状和模式以及神经生物素标记进行表征。2. 采用膜片钳技术研究全细胞电流对KATP通道开放剂二氮嗪(0.3 mM)以及KATP通道抑制剂甲苯磺丁脲(0.5 mM)或格列本脲(20 μM)的敏感性。全细胞电流被二氮嗪激活并被甲苯磺丁脲抑制的细胞比例在锥体细胞中为26%,在中间神经元中为89%,在胶质细胞中为100%,在颗粒细胞中为89%。二氮嗪诱导电流的反转电位处于钾离子平衡电位,当用无ATP的移液管溶液透析细胞时,类似的电流会自发激活。3. 使用单细胞逆转录聚合酶链反应(RT-PCR)方法,检测了CA1锥体细胞和中间神经元中编码KATP通道亚基(Kir6.1、Kir6.2、SUR1和SUR2)的mRNA的存在情况。仅在17%的锥体细胞中检测到可导致功能性KATP通道的亚基mRNA组合(Kir6.1与SUR1、Kir6.2与SUR1或SUR2)。另一方面,75%的中间神经元中可能形成KATP通道,主要是由Kir6.2与SUR1组合形成(占所有中间神经元的58%)。4. 这些综合分析结果表明,功能性KATP通道存在于大鼠海马体的主要神经元、中间神经元和胶质细胞中,但在所研究的四种细胞类型中密度差异很大。

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