Gautam Shree Hari, Otsuguro Ken-Ichi, Ito Shigeo, Saito Toshiyuki, Habara Yoshiaki
Laboratory of Physiology, Department of Biomedical Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan.
Neurosci Res. 2007 Jan;57(1):129-39. doi: 10.1016/j.neures.2006.09.016. Epub 2006 Oct 30.
T-type Ca(2+) channels are low-voltage-activated Ca(2+) channels that control Ca(2+) entry in excitable cells during small depolarization above resting potentials. Using Ca(2+) imaging with a laser scanning confocal microscope we investigated the involvement of T-type Ca(2+) channels in IBMX/forskolin- and sparingly elevated extracellular K(+)-induced Ca(2+) transients in freshly isolated porcine olfactory receptor neurons (ORNs). In the presence of mibefradil (10microM) or Ni(2+) (100microM), the selective T-type Ca(2+) channel inhibitors, IBMX/forskolin-induced Ca(2+) transients in the soma were either strongly (>60%) inhibited or abolished completely. However, the Ca(2+) transients in the knob were only partially (<60%) inhibited. Ca(2+) transients induced by 30mM K(+) were also partially ( approximately 60%) inhibited at both the knob and soma. Furthermore, ORNs responded to as little as a 2.5mM increase in the extracellular K(+) concentration (7.5mM K(+)), and such responses were completely inhibited by mibefradil or Ni(2+). These results reveal functional expression of T-type Ca(2+) channels in porcine ORNs, and suggest a role for these channels in the spread Ca(2+) transients from the knob to the soma during activation of the cAMP cascade following odorant binding to G-protein-coupled receptors on the cilia/knob of ORNs.
T型Ca(2+)通道是低电压激活的Ca(2+)通道,在静息电位之上的小去极化过程中控制可兴奋细胞中Ca(2+)的内流。我们使用激光扫描共聚焦显微镜进行Ca(2+)成像,研究了T型Ca(2+)通道在新分离的猪嗅觉受体神经元(ORN)中由异丁基甲基黄嘌呤(IBMX)/福斯高林以及细胞外K(+)浓度轻度升高所诱导的Ca(2+)瞬变中的作用。在存在米贝地尔(10μM)或Ni(2+)(100μM)这两种选择性T型Ca(2+)通道抑制剂的情况下,IBMX/福斯高林诱导的胞体Ca(2+)瞬变要么被强烈抑制(>60%),要么完全消除。然而,纤毛末端的Ca(2+)瞬变仅被部分抑制(<60%)。30mM K(+)诱导的Ca(2+)瞬变在纤毛末端和胞体也都被部分抑制(约60%)。此外,ORN对细胞外K(+)浓度仅增加2.5mM(至7.5mM K(+))就有反应,而这种反应被米贝地尔或Ni(2+)完全抑制。这些结果揭示了T型Ca(2+)通道在猪ORN中的功能性表达,并表明这些通道在气味剂与ORN纤毛/纤毛末端上的G蛋白偶联受体结合后激活cAMP级联反应期间,在Ca(2+)瞬变从纤毛末端扩散到胞体的过程中发挥作用。