Modulation and Resistance of Levamisole Receptor Channels

蛔虫病和钩虫感染影响着全世界16亿人。驱虫药，包括 左旋咪唑和相关药物(吡喃酮)，用于对抗线虫寄生虫，耐药性是一种威胁。 我们的长期目标是通过以下方式提高驱虫药的疗效，以改善人类健康 确定逆转耐药性的方法。此应用程序的目标是测试单通道 左旋咪唑受体的特性和描述线虫对 左旋咪唑和改变效力。我们的中心假设是L亚型的结构和孔隙 线虫肌肉上的乙酰胆碱受体离子通道使其对左旋咪唑和更多的 对钙的通透性；以及乙酰胆碱通道的反应增加(调制) 神经肽AF2参与cAMP、钙内流和激酶活性。这项研究的基本原理是，由于 调节对左旋咪唑激活的受体通道的反应的机制变得已知， 可以制定药理学方法来克服耐药性。我们将使用肌肉制剂 利用电流钳、电压钳和膜片钳技术对猪链霉菌、线虫和零突变体进行检测 L亚型乙酰胆碱通道的钙通透性和亚基组成及AF2模型的验证 调制。我们将追求三个目标：1)测定N亚型、L亚型和B亚型猪瘟病菌的钙渗透性 肌乙酰胆碱受体，从而确定首选靶点；2)在膜片钳中确定 在线虫中的实验，L亚型乙酰胆碱通道的亚基需求；3)表征， NA.suum肌肉中钙和AF2影响不同肌细胞开放的机制和药理学 Achr通道亚型，以增加胆碱能驱虫药的反应和效力。这个 研究是创新的，因为我们正在将线虫的新知识与先进的 线虫寄生的电生理学，包括用于膜片钳记录的肌泡准备。 我们希望这项研究能找到更多提高胆碱能效力的策略 驱虫药。这项研究意义重大，因为结果的应用将导致新的方法 控制和克服对左旋咪唑类驱虫药的耐药性。

Ascariasis and hookworm infection affect 1.6 billion people across the world. Anthelmintics, including levamisole and related drugs (pyrantel), are used to combat nematode parasites, and resistance is a threat. Our long-range objective is to improve human health by increasing the efficacy of anthelmintic drugs by identifying approaches to reverse resistance. The objective of this application is to test single-channel properties of levamisole receptors and a model that describes changes in the sensitivity of nematodes to levamisole and alters potency. Our central hypothesis is that the structure and pore of the L-subtype acetylcholine receptor ion-channel on nematode muscle makes it more sensitive to levamisole and more permeable to Ca; and the increased response of acetylcholine channels (modulation) produced by the neuropeptide, AF2,involves cAMP, Ca entry and kinase activity. The rationale for the research is that, as the mechanisms for modulating responses to levamisole activated receptor channels become known, pharmacological approaches can be formulated to overcome resistance. We will use muscle preparations of A. suum, C. elegans and null-mutants with current-clamp, voltage-clamp and patch-clamp technology to test the Ca permeability and subunit composition of L-subtype acetylcholine channels and to test a model for AF2 modulation. We will pursue 3 aims: 1) determine the Ca permeability of N-, L- and B- subtypes of A. suum muscle acetylcholine receptors and thereby identify a preferred target site; 2) determine in patch-clamp experiments in C. elegans, the subunit requirements of the L-subtype acetylcholine channel; 3) characterize, n A. suum muscle, the mechanism & pharmacology by which calcium and AF2 affect the opening of different AChR channel subtypes, in order to increase responses and potency of cholinergic anthelmintics. The research is innovative because we are combining new knowledge from C. elegans with advanced electrophysiology of nematode parasites including muscle-vesicle preparations for patch-clamp recordings. We expect the research to identify additional strategies that increase the potency of cholinergic anthelmintics. The research is significant because application of the results will to lead to new approaches to control and overcome resistance to anthelmintics of the levamisole class.