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豚鼠小肠培养肌间神经元中的P2X嘌呤受体

P2X purinoceptors in cultured myenteric neurons of guinea-pig small intestine.

作者信息

Zhou X, Galligan J J

机构信息

Department of Pharmacology and Toxicology, Michigan State University, East Lansing, USA.

出版信息

J Physiol. 1996 Nov 1;496 ( Pt 3)(Pt 3):719-29. doi: 10.1113/jphysiol.1996.sp021722.

DOI:10.1113/jphysiol.1996.sp021722
PMID:8930839
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1160859/
Abstract
  1. Fast excitatory postsynaptic currents (fEPSCs) and responses to exogenously applied purinoceptor agonists were studied in primary cultures of myenteric neurons from guinea-pig small intestine. Whole-cell and outside-out configurations of the patch clamp technique were used. Hexamethonium (100 microM) partly inhibited fEPSCs in 28% of neurons. Hexamethonium-resistant fEPSCs were inhibited by 97 +/- 2% by the P2X receptor antagonist, pyridoxal-phosphate-6-azophenyl-2',4'-disulphonic acid (PPADS, 10 microM). 2. ATP caused two types of inward currents. In 92% of neurons (n = 123), ATP caused a slowly desensitizing current that declined with a double exponential time course (tau 1 = 7.1 +/- 2.0 s; tau 2 = 57 +/- 7.4 s, n = 4). The rank order potency for purinoceptor agonists in these neurons was ATP > 2-methylthio-ATP (2-MeSATP) > > alpha, beta-methylene ATP (alpha, beta-me ATP) > beta, gamma-meATP > ADP. The EC50 values for ATP and 2-MeSATP were 40 and 65 microM, respectively. alpha, beta-MeATP acted as a partial agonist at these receptors. In 8% of neurons (n = 11), ATP-induced currents desensitized rapidly with a double exponential time course (tau 1 = 0.13 +/- 0.015 s; tau 2 = 2.2 +/- 1.3 s, n = 4); alpha, beta-meATP caused similar responses in these cells. Both types of ATP-induced current were associated with an increased conductance and an inwardly rectifying I-V relationship (Erev = 10 mV). Halving [Na+]o shifted the reversal potential of ATP currents by -22 +/- 6 mV. 3. ATP activated single channel currents in outside-out patches. The single channel I-V relationship was linear between -120 and 60 mV (Erev approximately 0 mV). Single channel conductance between -100 and -60 mV was 25 +/- 2 pS. Single channel open probability was voltage dependent and decreased from 0.05 +/- 0.01 at -100 mV to 0.007 +/- 0.002 at +40 mV. 4. These data show that P2X purinoceptors mediate some fEPSCs in cultured myenteric neurons. Myenteric neurons express the fast-desensitizing alpha, beta-me ATP-sensitive subtype of P2X receptor that has the properties of cloned P2X1 receptors and is similar to native receptors in smooth muscle cells. Myenteric neurons also express a P2X receptor that desensitized slowly and was alpha, beta-meATP-insensitive. This receptor has the properties of cloned P2X2 or P2X5 receptors and is similar to native receptors found in PC-12 cells and superior cervical ganglion neurons. The known distribution of P2X2 and P2X5 receptors suggests that myenteric neurons are likely to express predominantly P2X2 receptors.
摘要
  1. 在豚鼠小肠肌间神经元的原代培养物中研究了快速兴奋性突触后电流(fEPSCs)以及对外源性嘌呤受体激动剂的反应。采用了膜片钳技术的全细胞和外向膜片配置。六甲铵(100 microM)在28%的神经元中部分抑制fEPSCs。P2X受体拮抗剂磷酸吡哆醛 - 6 - 偶氮苯 - 2',4'-二磺酸(PPADS,10 microM)可使六甲铵抗性fEPSCs受到97±2%的抑制。

  2. ATP引起两种内向电流。在92%的神经元(n = 123)中,ATP引起一种缓慢脱敏的电流,其以双指数时间进程下降(τ1 = 7.1±2.0秒;τ2 = 57±7.4秒,n = 4)。这些神经元中嘌呤受体激动剂的效价顺序为ATP>2 - 甲硫基 - ATP(2 - MeSATP)>>α,β - 亚甲基ATP(α,β - me ATP)>β,γ - meATP>ADP。ATP和2 - MeSATP的EC50值分别为40和65 microM。α,β - MeATP在这些受体上起部分激动剂的作用。在8%的神经元(n = 11)中,ATP诱导的电流以双指数时间进程快速脱敏(τ1 = 0.13±0.015秒;τ2 = 2.2±1.3秒,n = 4);α,β - meATP在这些细胞中引起类似反应。两种类型的ATP诱导电流均与电导增加和内向整流的I - V关系(Erev = 10 mV)相关。将[Na+]o减半使ATP电流的反转电位偏移 - 22±6 mV。

  3. ATP在外向膜片中激活单通道电流。单通道I - V关系在 - 120至60 mV之间呈线性(Erev约为0 mV)。 - 100至 - 60 mV之间的单通道电导为25±2 pS。单通道开放概率依赖于电压,从 - 100 mV时的0.05±0.01降至 + 40 mV时的0.007±0.002。

  4. 这些数据表明,P2X嘌呤受体介导培养的肌间神经元中的一些fEPSCs。肌间神经元表达快速脱敏的α,β - me ATP敏感型P2X受体亚型,其具有克隆的P2X1受体的特性,且与平滑肌细胞中的天然受体相似。肌间神经元还表达一种缓慢脱敏且对α,β - meATP不敏感的P2X受体。该受体具有克隆的P2X2或P2X5受体的特性,且与PC - 12细胞和颈上神经节神经元中的天然受体相似。P2X2和P2X5受体的已知分布表明,肌间神经元可能主要表达P2X2受体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/15ff/1160859/707a20815ac6/jphysiol00391-0129-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/15ff/1160859/707a20815ac6/jphysiol00391-0129-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/15ff/1160859/707a20815ac6/jphysiol00391-0129-a.jpg

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