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蝾螈离体嗅觉受体细胞中细胞内环状AMP诱导的反应。

The response induced by intracellular cyclic AMP in isolated olfactory receptor cells of the newt.

作者信息

Kurahashi T

机构信息

Department of Information Physiology, National Institute for Physiological Sciences, Okazaki, Japan.

出版信息

J Physiol. 1990 Nov;430:355-71. doi: 10.1113/jphysiol.1990.sp018295.

Abstract
  1. Responses induced by intracellular cyclic nucleotides were analysed in isolated olfactory receptor cells of the newt under a voltage-clamp condition by using the patch pipette in a whole-cell recording configuration. Cyclic nucleotides were applied by diffusion from the patch pipette. 2. Introduction of either cyclic AMP or cyclic GMP caused a transient inward current in cells held at -50 mV. The response amplitude was dose-dependent with the Hill coefficient of 3 and half-saturating concentration of 300 microM (concentration in the pipette) for both cyclic AMP and cyclic GMP. Cyclic CMP was less effective than those two nucleotides. 3. The response to intracellular cyclic AMP was seen in all cilia-bearing cells, but not in cells which lost the cilia during dissociation. The response latency was shorter when cyclic AMP was introduced into the ciliated terminal swelling (ca 0.2 s) rather than into the cell body (ca 1.4 s). These results suggest that the sensitivity to intracellular cyclic AMP is confined to the cilia. 4. The cyclic AMP-induced current was transient (half decay time, ca 2.3s) despite the fact that cyclic AMP was continuously loaded from the patch pipette. The response time course was controlled by Ca2+; the reduction of external Ca2+ concentration (replaced with Mg2+) or loading the cell with 50 mM-EGTA prolonged the cyclic AMP-induced responses. The Ca2(+)-induced suppression was reversible. 5. The reversal potential of the cyclic AMP-induced transient current was -4.8 +/- 3.8 mV, and that of the current re-induced by Ca2+ removal was 1.5 +/- 2.1 mV, suggesting that both currents flowed through the same ionic channel. The channel permeates all alkali metal ions with the permeability ratios of PLi:PNa:PK:PRb:PCs = 0.93:1:0.93:0.91:0.72, but not Cl- or choline ions. 6. These results demonstrate that the cyclic AMP-induced response and the odorant-induced response of the isolated olfactory cell have nearly identical characteristics. The present study supports the notion that cyclic AMP is the internal messenger mediating olfactory transduction.
摘要
  1. 在电压钳制条件下,采用膜片吸管全细胞记录模式,对蝾螈分离的嗅觉受体细胞中细胞内环状核苷酸诱导的反应进行了分析。环状核苷酸通过从膜片吸管扩散施加。2. 引入环磷酸腺苷(cAMP)或环磷酸鸟苷(cGMP)会在保持在 -50 mV 的细胞中引起瞬时内向电流。对于 cAMP 和 cGMP,反应幅度呈剂量依赖性,希尔系数为 3,半饱和浓度为 300 μM(吸管中的浓度)。环磷酸胞苷(cCMP)的效果不如这两种核苷酸。3. 在所有带有纤毛的细胞中都能观察到对细胞内 cAMP 的反应,但在解离过程中失去纤毛的细胞中则没有。当将 cAMP 引入纤毛状末端膨大处(约 0.2 秒)而非细胞体(约 1.4 秒)时,反应潜伏期更短。这些结果表明,对细胞内 cAMP 的敏感性局限于纤毛。4. 尽管 cAMP 从膜片吸管持续加载,但 cAMP 诱导的电流是瞬时的(半衰期约 2.3 秒)。反应时间进程受 Ca2+ 控制;降低外部 Ca2+ 浓度(用 Mg2+ 替代)或用 50 mM - 乙二醇双四乙酸(EGTA)加载细胞会延长 cAMP 诱导的反应。Ca2+ 诱导的抑制是可逆的。5. cAMP 诱导的瞬时电流的反转电位为 -4.8 ± 3.8 mV,去除 Ca2+ 后重新诱导的电流的反转电位为 1.5 ± 2.1 mV,这表明两种电流通过相同的离子通道。该通道可通透所有碱金属离子,通透率之比为 PLi:PNa:PK:PRb:PCs = 0.93:1:0.93:0.91:0.72,但不通透 Cl- 或胆碱离子。6. 这些结果表明,分离的嗅觉细胞中 cAMP 诱导的反应和气味剂诱导的反应具有几乎相同的特征。本研究支持 cAMP 是介导嗅觉转导的细胞内信使这一观点。

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