Xia Jingsheng, Pan Rong, Gao Xinghua, Meucci Olimpia, Hu Huijuan
Department of Pharmacology and Physiology, Drexel University College of Medicine, Philadelphia, USA.
Department of Pharmacology and Physiology, Drexel University College of Medicine, Philadelphia, USA Department of Pharmacology of Chinese Materia Medica, China Pharmaceutical University, Nanjing, China.
J Physiol. 2014 Aug 15;592(16):3443-61. doi: 10.1113/jphysiol.2014.275065. Epub 2014 May 23.
Store-operated calcium channels (SOCs) are calcium-selective cation channels that mediate calcium entry in many different cell types. Store-operated calcium entry (SOCE) is involved in various cellular functions. Increasing evidence suggests that impairment of SOCE is responsible for numerous disorders. A previous study demonstrated that YM-58483, a potent SOC inhibitor, strongly attenuates chronic pain by systemic or intrathecal injection and completely blocks the second phase of formalin-induced spontaneous nocifensive behaviour, suggesting a potential role of SOCs in central sensitization. However, the expression of SOCs, their molecular identity and function in spinal cord dorsal horn neurons remain elusive. Here, we demonstrate that SOCs are expressed in dorsal horn neurons. Depletion of calcium stores from the endoplasmic reticulum (ER) induced large sustained calcium entry, which was blocked by SOC inhibitors, but not by voltage-gated calcium channel blockers. Depletion of ER calcium stores activated inward calcium-selective currents, which was reduced by replacing Ca(2+) with Ba(2+) and reversed by SOC inhibitors. Using the small inhibitory RNA knockdown approach, we identified both STIM1 and STIM2 as important mediators of SOCE and SOC current, and Orai1 as a key component of the Ca(2+) release-activated Ca(2+) channels in dorsal horn neurons. Knockdown of STIM1, STIM2 or Orai1 decreased resting Ca(2+) levels. We also found that activation of neurokinin 1 receptors led to SOCE and activation of SOCs produced an excitatory action in dorsal horn neurons. Our findings reveal that a novel SOC signal is present in dorsal horn neurons and may play an important role in pain transmission.
储存性钙通道(SOCs)是钙选择性阳离子通道,介导多种不同细胞类型的钙内流。储存性钙内流(SOCE)参与多种细胞功能。越来越多的证据表明,SOCE受损是许多疾病的原因。先前的一项研究表明,强效SOC抑制剂YM-58483通过全身或鞘内注射可强烈减轻慢性疼痛,并完全阻断福尔马林诱导的自发伤害性防御行为的第二阶段,提示SOCs在中枢敏化中可能发挥作用。然而,SOCs在脊髓背角神经元中的表达、其分子特性和功能仍不清楚。在此,我们证明SOCs在背角神经元中表达。内质网(ER)钙储存的耗竭诱导了大量持续的钙内流,该内流被SOC抑制剂阻断,但未被电压门控钙通道阻滞剂阻断。ER钙储存的耗竭激活了内向钙选择性电流,用Ba(2+)替代Ca(2+)可使其降低,SOC抑制剂可使其逆转。使用小干扰RNA敲低方法,我们确定STIM1和STIM2都是SOCE和SOC电流的重要介质,而Orai1是背角神经元中钙释放激活钙通道(CRAC)的关键组成部分。敲低STIM1、STIM2或Orai1可降低静息钙水平。我们还发现,神经激肽1受体的激活导致SOCE,SOCs的激活在背角神经元中产生兴奋性作用。我们的研究结果表明,背角神经元中存在一种新的SOC信号,可能在疼痛传递中起重要作用。