公开(公告)号：US06358701B1

公开(公告)日：2002-03-19

申请号：US09130339

申请日：1998-08-06

申请人： Jeffrey W. Warmke ,  Doris F. Cully ,  Adrian Etter ,  Philip S. Paress ,  Charles J. Cohen ,  Richard Brochu

发明人： Jeffrey W. Warmke ,  Doris F. Cully ,  Adrian Etter ,  Philip S. Paress ,  Charles J. Cohen ,  Richard Brochu

IPC分类号： C07H2104

CPC分类号： C07K14/4359 ,  C07K14/70571

摘要： To date, L-glutamate-gated chloride (GluCl) channels have been observed only in invertebrate organisms. Modulators of this channel (either agonists or antagoinists) will interfere with neurotransmission. For example, agents such as avermectins activate the GluCl, causing paralysis due to blocking of neurotranmitter release, resulting in death of the organism. Because GluCl channels are invertebreate specific, they are excellent targets for the discovery of novel insecticides, anthelminths and parasiticides that will display a marked safety profile because of the lack of mechanism based toxicity in vertebrate organisms. The present specification discloses isolation of a cDNA clone from the cat flea Ctenocephalides felis (CfGluCl-1) that encodes a L-glutamate-gated chloride channel. Heterologous expression of CfGluCl-1 cRNA in Xenopus oocytes results in robust expression of a L-glutamate-gated chloride current and the channel is activated and potentiated by avermectins. The expression of CfGluCl-1 in a heterologous expression system if useful to screens for novel GluCl channel agonsts and antagonsits. Additionally, this specification disclose impoved methods of screening for GluCl channel modulators.

摘要翻译： 迄今为止，仅在无脊椎动物体内观察到L-谷氨酸氯化物（GluCl）通道。 该通道的调节剂（激动剂或antagoinists）将干扰神经传递。 例如，除虫菌素之类的试剂激活GluCl，由于神经递质释放的阻断导致麻痹，导致生物体的死亡。 由于GluCl通道具有特异性，因此它们是发现新型杀虫剂，anthelminths和杀寄生虫剂的优秀目标，由于脊椎动物体内缺乏基于机制的毒性，因此将显示出明显的安全性。 本说明书公开了从编码L-谷氨酸盐化氯化物通道的猫蚤Ctenocephalides felis（CfGluCl-1）中分离cDNA克隆。 非洲爪蟾卵母细胞中CfGluCl-1 cRNA的异源表达导致L-谷氨酸氯化物电流的强烈表达，并且通道被阿凡曼菌素激活和增强。 CfGluCl-1在异源表达系统中的表达如果可用于筛选新的GluCl通道激动剂和antagonsits。 另外，本说明书公开了筛选GluCl通道调节剂的方法。