Nakajo Koichi, Katsuyama You, Ono Fumihito, Ohtsuka Yukio, Okamura Yasushi
Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro-ku, Tokyo 153-8902, Japan.
Neurosci Res. 2003 Jan;45(1):59-70. doi: 10.1016/s0168-0102(02)00193-1.
Ascidians belong to the primitive chordates and their larvae show symmetrical beating of the tail, which is reminiscent of the swimming pattern in primitive vertebrates. Since ascidian larva contains only a small number of neurons in their entire larval nervous system, they will potentially provide a simple model for the study of animal locomotion. In a step towards the goal of establishing the molecular basis underlying ascidian larval neurophysiology, we describe here a Kv4 class of voltage-gated potassium channel, TuKv4, from Halocynthia roretzi. Whole mount in situ hybridization indicates that TuKv4 is expressed in most of larval neurons including motor neurons. TuKv4-currents reconstituted in Xenopus oocytes show currents with similar properties to the lower-threshold A-type currents from cleavage-arrested ascidian blastomeres of neural lineage. However, the voltage-dependency of the steady-state inactivation and activation was shifted leftward by 20 mV, as compared with native A-type currents, suggesting that other components may be required to restore full function of the Kv4 channel. Unexpectedly, another isoform lacking C-terminal cytoplasmic region was also isolated. This truncated isoform did not lead to a functional current in Xenopus oocytes. RT-PCR analysis showed that the truncated form is transiently expressed during larval development, suggesting some developmental role for potassium channel expression.
海鞘属于原始脊索动物,其幼虫的尾部呈现对称摆动,这让人联想到原始脊椎动物的游泳模式。由于海鞘幼虫的整个幼虫神经系统中仅含有少量神经元,它们有可能为动物运动研究提供一个简单的模型。为了朝着建立海鞘幼虫神经生理学分子基础这一目标迈进,我们在此描述了一种来自柄海鞘的电压门控钾通道Kv4家族成员TuKv4。整体原位杂交表明TuKv4在包括运动神经元在内的大多数幼虫神经元中表达。在非洲爪蟾卵母细胞中重组的TuKv4电流显示出与神经谱系的卵裂阻滞海鞘卵裂球的低阈值A型电流具有相似特性的电流。然而,与天然A型电流相比,稳态失活和激活的电压依赖性向左偏移了20 mV,这表明可能需要其他成分来恢复Kv4通道的完整功能。出乎意料的是,还分离出了另一种缺乏C末端胞质区域的异构体。这种截短的异构体在非洲爪蟾卵母细胞中未产生功能性电流。逆转录聚合酶链反应分析表明,截短形式在幼虫发育过程中短暂表达,提示钾通道表达具有某种发育作用。