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通过细胞内注射环磷酸腺苷使鲤鱼和乌龟视网膜中的水平细胞去耦联。

Decoupling of horizontal cells in carp and turtle retinae by intracellular injection of cyclic AMP.

作者信息

Miyachi E, Murakami M

机构信息

Department of Physiology, Keio University School of Medicine, Tokyo, Japan.

出版信息

J Physiol. 1989 Dec;419:213-24. doi: 10.1113/jphysiol.1989.sp017870.

Abstract
  1. Horizontal cells are electrically coupled through gap junctions. This is a disadvantage in elucidating the membrane properties of the cells. In order to block gap junctions, adenosine 3',5'-cyclic monophosphate (cyclic AMP) or its analogues, dibutyryl cyclic AMP and 8-bromo cyclic AMP, were ionophoretically injected into horizontal cells of the carp or turtle retina. 2. Before injection of the chemicals the input resistance of the cell was so low as to be unmeasurable, because the applied current leaked through gap junctions. After injection, however, the input resistance was significantly increased. 3. After the injection dye-coupling between horizontal cells was not observed when examined by intracellular injection of Lucifer Yellow dye, supporting the idea that high concentrations of intracellular cyclic AMP block gap junctions. 4. In this situation responses to light delivered to the receptive field centre were increased in amplitude, while responses to light delivered to the receptive field surround were greatly diminished. 5. After injection horizontal cells were readily polarized by conventional intracellular current injection. The hyperpolarizing light responses in carp and turtle luminosity-type cells (H1 cells) could be reversed by depolarizing the horizontal cells, and the reversal potentials were estimated to be about 0 mV. In addition, the resistance increase which accompanied the hyperpolarizing light responses could be detected. 6. In turtle biphasic chromaticity-type horizontal cells (H2 cells), hyperpolarizing light responses to shorter wavelengths and depolarizing ones to longer wavelengths could be reversed by depolarizing the horizontal cells. Both responses have almost the same reversal potential at about 0 mV. The membrane resistance changes associated with light responses were also detected; the resistance increased during the hyperpolarizing response, while it decreased during the depolarizing response. These observations suggest that the ionic mechanisms of both responses are probably the same, irrespective of their polarities.
摘要
  1. 水平细胞通过缝隙连接进行电耦合。这在阐明细胞的膜特性方面是一个不利因素。为了阻断缝隙连接,将3',5'-环磷酸腺苷(环化AMP)或其类似物二丁酰环化AMP和8-溴环化AMP离子导入鲤鱼或乌龟视网膜的水平细胞中。2. 在注入化学物质之前,细胞的输入电阻很低以至于无法测量,因为施加的电流通过缝隙连接泄漏。然而,注入后,输入电阻显著增加。3. 注入后,通过细胞内注入鲁米诺黄染料检测时,未观察到水平细胞之间的染料耦合,这支持了高浓度细胞内环化AMP阻断缝隙连接的观点。4. 在这种情况下,传递到感受野中心的光刺激所引发的反应幅度增加,而传递到感受野周边的光刺激所引发的反应则大大减弱。5. 注入后,水平细胞很容易被传统的细胞内电流注入极化。鲤鱼和乌龟亮度型细胞(H1细胞)中的超极化光反应可以通过使水平细胞去极化而反转,反转电位估计约为0 mV。此外,还可以检测到伴随超极化光反应的电阻增加。6. 在乌龟双相色度型水平细胞(H2细胞)中,对较短波长的超极化光反应和对较长波长的去极化光反应可以通过使水平细胞去极化而反转。两种反应在约0 mV处几乎具有相同的反转电位。还检测到与光反应相关的膜电阻变化;在超极化反应期间电阻增加,而在去极化反应期间电阻降低。这些观察结果表明,两种反应的离子机制可能相同,无论其极性如何。

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