Department of Physiology, Biophysics and Neuroscience, Center for Research and Advanced Studies of the National Polytechnic Institute (Cinvestav-IPN), Mexico City, CP 07300, Mexico.
Neuroscience. 2011 Sep 29;192:74-80. doi: 10.1016/j.neuroscience.2011.06.073. Epub 2011 Jul 1.
GABA(A) receptors mediating tonic inhibitory currents are present in neurons from hippocampus, cerebellum, sensory cortex and thalamus. These receptors located at peri- and extra-synaptic sites are constituted mainly by α(4/6) and α(5) subunits which confer them high affinity for GABA and low desensitization. Immunohistochemical and in vitro hybridization studies have shown the expression of these subunits, while functional studies have reported the presence of GABAergic tonic currents in spinal dorsal horn neurons. However, the presence of this inhibitory current has not been documented in motoneurons. In addition, we previously reported that the monosynaptic reflex is facilitated by furosemide, an antagonist of the α(4/6) GABA(A) receptors, without affecting the dorsal root potential, which suggests the presence of a GABAergic tonic inhibitory current in motoneurons. The aim of this work was to investigate the presence of high affinity GABA(A) receptors in motoneurons. By intracellular recordings made with sharp electrodes and the whole-cell patch clamp recording technique we show here that the membrane input resistance and the monosynaptic excitatory post-synaptic potential (EPSPs) are significantly increased by bicuculline. Likewise, the depression of the EPSPs and the input membrane resistance normally induced by muscimol was partially reverted by 20 μM bicuculline and abolished when the concentration of the antagonist was raised to 100 μM. Last, bicuculline at low concentration did not affect the holding current as occur with the high concentration that block the tonic inhibitory GABAergic current. Together these results suggest that the excitability in motoneurons may be tonically inhibited by high affinity GABA(A) receptors.
介导紧张性抑制电流的 GABA(A) 受体存在于海马体、小脑、感觉皮层和丘脑的神经元中。这些位于突触前和突触外部位的受体主要由 α(4/6) 和 α(5) 亚基组成,它们赋予其对 GABA 的高亲和力和低脱敏性。免疫组织化学和体外杂交研究表明了这些亚基的表达,而功能研究报告了脊髓背角神经元中存在 GABA 能紧张性电流。然而,这种抑制电流在运动神经元中尚未被记录到。此外,我们之前的研究报告表明,速尿作为 α(4/6) GABA(A) 受体的拮抗剂,可以促进单突触反射,而不影响背根电位,这表明运动神经元中存在 GABA 能紧张性抑制电流。本工作旨在研究运动神经元中是否存在高亲和力 GABA(A) 受体。通过使用尖锐电极进行细胞内记录和全细胞膜片钳记录技术,我们在这里显示,在高亲和力 GABA(A) 受体存在的情况下,膜输入电阻和单突触兴奋性突触后电位 (EPSPs) 被荷包牡丹碱显著增加。同样,由 muscimol 引起的 EPSPs 和输入膜电阻的抑制被 20 μM 荷包牡丹碱部分逆转,当拮抗剂的浓度升高到 100 μM 时,抑制被完全消除。最后,低浓度的荷包牡丹碱不会像高浓度那样影响保持电流,而高浓度会阻断紧张性抑制性 GABA 能电流。综上所述,这些结果表明,运动神经元的兴奋性可能受到高亲和力 GABA(A) 受体的紧张性抑制。