Tsunoda S, Salkoff L
Department of Anatomy and Neurobiology, Washington University School of Medicine, St. Louis, Missouri 63110.
J Neurosci. 1995 Mar;15(3 Pt 1):1741-54. doi: 10.1523/JNEUROSCI.15-03-01741.1995.
In this study, we perform the first genetic analysis of K+ currents in Drosophila embryonic neurons revealing the identity of the currents present. Unlike muscles, where the presence of Shaker is obvious, Shaker currents are not detectable in these neurons. In contrast, we show that Shal is as important in these neuronal cell bodies as Shaker is in muscles. Only three single-channel currents were found, all of which are genetically separable. Shal encodes a 4 pS transient channel. Whole-cell Shal currents have a wide variety of inactivation rates which, in contrast to a mechanism such as heteromultimer formation, is due to single Shal channels assuming different gating modes. Shaw encodes a 42 pS noninactivating channel distinctive for its extremely low voltage sensitivity; Shaw channels have a total equivalent gating charge of 0.90 e- charges, in sharp contrast to 7 e- reported for Shaker channels. An 11 pS slowly inactivating channel also present in these neurons may be encoded by the Shab gene. Thus, of four voltage-dependent K+ channel genes now cloned in Drosophila, all except Shaker appear to be expressed in the cell bodies of these neurons. It is only in Drosophila that a study such as this one can be done. Because Drosophila contains only a single member of each of the four subfamilies of K+ channel genes (unlike mammals), we can eliminate an entire subfamily with a mutation to a single gene. Here, we have examined the effect of eliminating each of three subfamilies. Such a task is presently impossible to accomplish in any mammalian system.
在本研究中,我们对果蝇胚胎神经元中的钾离子电流进行了首次基因分析,揭示了所存在电流的特性。与肌肉中明显存在Shaker不同,在这些神经元中检测不到Shaker电流。相比之下,我们发现Shal在这些神经元胞体中的重要性与Shaker在肌肉中的重要性相当。仅发现三种单通道电流,所有这些电流在基因上都是可分离的。Shal编码一个4 pS的瞬时通道。全细胞Shal电流具有多种失活速率,这与诸如异源多聚体形成等机制不同,是由于单个Shal通道呈现不同的门控模式所致。Shaw编码一个42 pS的非失活通道,其独特之处在于电压敏感性极低;Shaw通道的总等效门控电荷为0.90个电子电荷,与报道的Shaker通道的7个电子电荷形成鲜明对比。这些神经元中还存在的一个11 pS的缓慢失活通道可能由Shab基因编码。因此,在果蝇中现已克隆的四个电压依赖性钾离子通道基因中,除了Shaker之外,其他基因似乎都在这些神经元的胞体中表达。只有在果蝇中才能进行这样的研究。因为果蝇每个钾离子通道基因亚家族仅包含一个成员(与哺乳动物不同),我们可以通过单个基因突变消除整个亚家族。在此,我们研究了消除三个亚家族中每个亚家族的影响。目前在任何哺乳动物系统中都无法完成这样的任务。