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)通道基因亚家族的特性 KCNG。电压门控离子通道在决定 神经的电化学性质和由此产生的功能， 神经感觉和兴奋性细胞。我们的初步结果表明 KCNG(KV7)亚家族中至少有两个成员。的表达 KCNG1基因在耳蜗(内毛细胞和外毛细胞)中观察到， 在前庭和螺旋神经节细胞亚群中， 大脑的区域(皮质、海马体、黑质和脑 茎)，以及在心脏组织中。我们假设KCNG1基因扮演着一个 在通过与兴奋性的有组织的联系来听力方面的重要作用 细胞，以及耳朵和大脑的神经和神经感觉组织。我们 计划通过以下特征来验证这一假设：1)KCNG基因亚家族 成员；2)KV7通道的结构/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。因此，我们计划分离和鉴定Kv7.1基因和 通过初步确定其染色体来确定其相关的亚家族成员 通过荧光原位杂交(FISH)定位，然后 确定它们的基因组序列、基因组织和基因 利用双脱氧核苷酸测序技术研究EMBL3的调控元件 人类淋巴细胞基因组克隆。此外，大小和组织分布 的成绩单将由Northern分析进行审查。序列数据和 关于KV7转录本异质性的信息，交替诱导 剪接的异构体，将通过筛选cDNA文库和 采用逆转录聚合酶链式反应和RACE-聚合酶链式反应。HKcng1α亚基和嵌合体 通道将在非洲爪哇卵母细胞中表达，其结构/功能 将通过检查它们的药理和 电生理特性。将进行实验，以生产和生产 评估mKCNG1基因敲除小鼠。Kv7.1基因敲除表型的检测 将有助于确定KCNG1在听觉中的生理作用 系统。我们还将研究两国之间可能存在的关系 人类KCNG1基因功能障碍(S)与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 其药理性质与IA电流相似，但速度较慢 以及非常小的失活。该电流可能代表功能Kcng1 频道。 总而言之，这些研究将确定 HKCNG基因在神经感觉和神经组织中的表达，以及定义 电压门控KCNG离子通道基因与听觉的关系 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.