Löhrke S, Kungel M, Friauf E
Physiologie III, Universität Frankfurt, Germany.
J Neurobiol. 1998 Sep 5;36(3):395-409.
The medial nucleus of the trapezoid body (MNTB) is a conspicuous structure in the mammalian auditory brain stem. It is a major component of the superior olivary complex and is involved in sound localization. Recently, organotypic slice culture preparations of the superior olivary complex were introduced to investigate the development of inhibitory and excitatory projections (Sanes and Hafidi, 1996; Lohmann et al., 1998). In the present article, we further assessed the organotypicity of our culture system (Lohmann et al., 1998) and examined electrical membrane properties of MNTB neurons expressed under culture conditions. To do so, MNTB neurons from early postnatal rats (P3-5) were studied after 3-6 days in vitro (DIV) by whole-cell patch-clamp recordings. Their mean resting potential was -59 mV, the input resistance averaged 171 Momega, and the average time constant was 3 ms. Four types of voltage-activated conductances were observed in voltage-clamp recordings. All cells expressed a tetrodotoxin (TTX)-sensitive sodium current. Two types of potassium currents could be characterized: a tetraethylammonium (TEA) -sensitive and a 4-aminopyridine (4-AP)-sensitive conductance, both of which are composed of a transient and a sustained component. Finally, an inwardly rectifying current, activated by hyperpolarizing voltage steps, was found. In current-clamp recordings, depolarizing current pulses typically elicited a single action potential. In the presence of 4-AP, however, these current pulses induced a train of action potentials. The duration of action potentials was increased by 4-AP and the afterhyperpolarization was reduced. Hyperpolarizing current injections induced a "sag" in the membrane potential, indicating the influence of an inwardly rectifying current. Our results demonstrate that MNTB neurons in slice cultures have electrical membrane properties comparable to those of their counterparts in acute slices.
梯形体内侧核(MNTB)是哺乳动物听觉脑干中的一个显著结构。它是上橄榄复合体的主要组成部分,参与声音定位。最近,上橄榄复合体的器官型切片培养制剂被引入以研究抑制性和兴奋性投射的发育(Sanes和Hafidi,1996;Lohmann等人,1998)。在本文中,我们进一步评估了我们培养系统的器官型特征(Lohmann等人,1998),并研究了在培养条件下表达的MNTB神经元的电膜特性。为此,通过全细胞膜片钳记录,对出生后早期大鼠(P3 - 5)的MNTB神经元在体外培养3 - 6天(DIV)后进行了研究。它们的平均静息电位为 -59 mV,输入电阻平均为171 MΩ,平均时间常数为3 ms。在电压钳记录中观察到四种类型的电压激活电导。所有细胞都表达对河豚毒素(TTX)敏感的钠电流。可以鉴定出两种类型的钾电流:对四乙铵(TEA)敏感和对4 - 氨基吡啶(4 - AP)敏感的电导,两者均由一个瞬态成分和一个持续成分组成。最后,发现了一种由超极化电压阶跃激活的内向整流电流。在电流钳记录中,去极化电流脉冲通常引发单个动作电位。然而,在存在4 - AP的情况下,这些电流脉冲诱导出一串动作电位。动作电位的持续时间因4 - AP而增加,超极化后电位减小。超极化电流注入在膜电位中诱导出“下垂”,表明存在内向整流电流的影响。我们的结果表明,切片培养中的MNTB神经元具有与急性切片中对应神经元相当的电膜特性。