NOVEL KV GENES, KCNG--CHARACTERIZATION AND DISEASE ASSOCIATION

我们的长期目标是确定结构和功能 一个新的电压门控K+离子（Kv）通道基因亚家族的性质， KCNG。电压门控离子通道在决定细胞的增殖中起着不可或缺的作用。 电化学性质和由此产生的神经功能， 神经感觉细胞和兴奋细胞。 我们的初步结果表明， 在KCNG（Kv 7）亚家族中存在至少两个成员。表达 在耳蜗（内毛细胞和外毛细胞）中观察到KCNG 1基因， 在前庭和螺旋神经节细胞亚群中， 大脑区域（皮质、海马、黑质和大脑） 干）和心脏组织中。我们假设KCNG 1基因在 通过与兴奋性神经元的协调联系， 细胞，以及耳和脑的神经和神经感觉组织。我们 我计划通过以下特征来验证这一假设：1）KCNG基因亚家族 Kv 7通道的结构与功能关系 蛋白质; 3）Kv7.1基因在听觉系统中的作用;以及4） 该基因与神经、神经肌肉和神经感觉的关系 功能协调发展的因此，我们计划分离和表征Kv7.1基因， 其相关的亚家族成员通过最初确定其染色体 通过荧光原位杂交（FISH）定位，然后 确定他们的基因组序列，基因组织， 通过EMBL 3 λ的双脱氧核苷酸测序的调节元件 人淋巴细胞基因组克隆。此外，大小和组织分布 将通过北方分析来检查转录本。序列数据和 关于Kv 7转录本异质性的信息， 剪接的同种型，将通过筛选cDNA文库获得， 采用RT-PCR和RACE-PCR技术。hKcng 1 α亚基和嵌合 通道将在非洲爪蟾卵母细胞中表达， 将通过检查它们的药理学和 电生理特性实验将进行生产和 评估mKCNG 1敲除小鼠。Kv7.1敲除表型的检查 将有助于确定KCNG 1在听觉系统中的生理作用， 系统我们还将研究 功能失调的人类KCNG 1基因和Jervell Lange-Niesen家族。 最近，我们的同事Bertrand博士发现了一种新的OHC K+电流， 其药理学性质类似于IA电流，但具有缓慢的 和非常小的失活。该电流可以表示函数Kcng 1 渠道 总的来说，这些研究将确定 hKCNG基因在感觉神经和神经组织中的表达，以及定义 电压门控性KCNG离子通道基因与听觉神经元功能的关系 系统

Our long term goal is to determine the structural and functional properties of a new voltage-gated K+ ion (Kv) channel gene subfamily, KCNG. Voltage-gated ion channels play an integral role in determining the electrochemical properties and resulting functions of neural, neurosensory, and excitatory cells. Our preliminary results suggest that there are at least two members in the KCNG (Kv7) subfamily. Expression of the KCNG1 gene was observed in the cochlea (inner and outer hair cells), in subpopulations of vestibular and spiral ganglion cells, in specific areas of the brain (cortex, hippocampus, substantia nigra, and brain stem), and in cardiac tissue. We hypothesize that the KCNG1 gene plays a vital role in hearing through an orchestrated association with excitatory cells, as well as neural and neurosensory tissues of the ear and brain. We plan to test this hypothesis by characterizing: 1) the KCNG gene subfamily members; 2) the structure/function relationships of the Kv7 channel proteins; 3) the role of the Kv7.1 gene in the auditory system; and 4) the relationship of this gene with neural, neuromuscular, and neurosensory functions. Thus, we plan to isolate and characterize the Kv7.1 gene and its related subfamily member by initially determining their chromosomal locations through fluorescence in situ hybridization (FISH), and then identifying their genomic sequences, gene organization, and genetic regulatory elements by dideoxynucleotide sequencing of the EMBL3 lambda human lymphocyte genomic clones. In addition, size and tissue distribution of transcripts will be examined by Northern analyses. Sequence data and information about Kv7 transcript heterogeneity, inducing alternatively spliced isoforms, will be obtained by screening cDNA libraries and employing RTPCR and RACE-PCR. The hKcng1 alpha subunits and chimeric channels will be expressed in Xenopus oocytes, and the structure/function relationships will be studied by examining their pharmacological and electrophysiological properties. Experiments will be done to produce and assess mKCNG1 knock out mice. Examination of the Kv7.1 knock out phenotype will assist in determining the physiological role of KCNG1 in the auditory system. We will also examine the possible relationship between the dysfunctional human KCNG1 gene(s) and the Jervell Lange-Niesen families. Recently, our colleague Dr. Bertrand has identified a novel OHC K+ current whose pharmacological properties resemble the IA current but with a slow and very small inactivation. This current may represent functional Kcng1 channels. Collectively, these studies will determine the function and role of the hKCNG genes in neurosensory and neural tissue, as well as define the relationship of voltage-gated KCNG ion channel genes with the auditory system.