Sharma Kushal, Kang Kwon Woo, Seo Young-Woo, Glowatzki Elisabeth, Yi Eunyoung
College of Pharmacy and Natural Medicine Research Institute, Mokpo National University, Muan 58554, Korea.
KBSI Gwangju Center, Korea Basic Science Institute, Gwangju 61186, Korea.
Exp Neurobiol. 2022 Aug 31;31(4):243-259. doi: 10.5607/en22013.
Cochlear afferent nerve fibers (ANF) are the first neurons in the ascending auditory pathway. We investigated the low-voltage activating K channels expressed in ANF dendrites using isolated rat cochlear segments. Whole cell patch clamp recordings were made from the dendritic terminals of ANFs. Outward currents activating at membrane potentials as low as -64 mV were observed in all dendrites studied. These currents were inhibited by 4-aminopyridine (4-AP), a blocker known to preferentially inhibit low-voltage activating K currents (I) in CNS auditory neurons and spiral ganglion neurons. When the dendritic I was blocked by 4-AP, the EPSP decay time was significantly prolonged, suggesting that dendritic I speeds up the decay of EPSPs and likely modulates action potentials of ANFs. To reveal molecular subtype of dendritic I, α-dendrotoxin (α-DTX), a selective inhibitor for K1.1, K1.2, and K1.6 containing channels, was tested. α-DTX inhibited 23±9% of dendritic I. To identify the α-DTXsensitive and α-DTX-insensitive components of I, immunofluorescence labeling was performed. Strong K1.1- and K1.2-immunoreactivity was found at unmyelinated dendritic segments, nodes of Ranvier, and cell bodies of most ANFs. A small fraction of ANF dendrites showed K7.2- immunoreactivity. These data suggest that dendritic I is conducted through K1.1and K1.2 channels, with a minor contribution from K7.2 and other as yet unidentified channels.
耳蜗传入神经纤维(ANF)是听觉上行通路中的第一级神经元。我们使用分离的大鼠耳蜗节段研究了ANF树突中表达的低电压激活钾通道。对ANF的树突末梢进行全细胞膜片钳记录。在所有研究的树突中均观察到外向电流在低至 -64 mV的膜电位时激活。这些电流被4-氨基吡啶(4-AP)抑制,4-AP是一种已知优先抑制中枢神经系统听觉神经元和螺旋神经节神经元中低电压激活钾电流(I)的阻滞剂。当树突I被4-AP阻断时,兴奋性突触后电位(EPSP)的衰减时间显著延长,这表明树突I加速了EPSP的衰减,并且可能调节ANF的动作电位。为了揭示树突I的分子亚型,测试了α-银环蛇毒素(α-DTX),它是一种对包含K1.1、K1.2和K1.6的通道具有选择性的抑制剂。α-DTX抑制了23±9%的树突I。为了鉴定I中对α-DTX敏感和不敏感的成分,进行了免疫荧光标记。在大多数ANF的无髓鞘树突段、郎飞结和细胞体中发现了强烈的K1.1和K1.2免疫反应性。一小部分ANF树突显示出K7.2免疫反应性。这些数据表明,树突I是通过K1.1和K1.2通道传导的,K7.2和其他尚未确定的通道贡献较小。