謝汝敦2006-07-262018-07-112006-07-262018-07-112004http://ntur.lib.ntu.edu.tw//handle/246246/25517氯離子通道在血管平滑肌的電生理學上扮演重要的角色。在包括血管的許多組織上給 予Agonist，可以誘發出氯離子電流。最近的研究顯示無論使用干擾氯離子穿越細胞膜或 阻斷氯離子通道的方法，都可以降低新腎上腺素對血管平滑肌產生的收縮。而陰莖海綿體 擁有獨特的膨脹與消脹的能力，基於陰莖海綿體與血管在構造與藥理學上的相似性，我們 可以預測氯離子對調節陰莖海綿體平滑肌張力﹝也就是勃起功能﹞扮演一定的角色。由先 前的計畫得知CLC-3 氯離子通道蛋白質是存在於老鼠陰莖海綿體平滑肌細胞及組織上的， 進一步的免疫組織化學分析與Northern Blot 分析將有助於進一步了解CLC-3 氯離子通道 在海綿體平滑肌上的分子生物學表現。實驗的內容包括 1. 免疫組織化學分析 探測老鼠的海綿體平滑肌組織上CLC-3 氯離子通道蛋白質的分佈情形。 2. 探針的製備 使用PCR 放大的方法來製備探針. 3. Northern Blot 分析 探測老鼠的海綿體平滑肌組織與細胞上CLC-3 氯離子通道基因的表現. 實驗結果得知使用免疫組織化學的分析的確可以在老鼠的海綿體平滑肌組織上偵測到 CLC-3 氯離子通道蛋白質的存在。而使用PCR 放大的方法亦可以製備出要用的探針。Chloride ion currents play a critical role in vascular smooth muscle electrophysiology. Agonist-induced, inward chloride currents have now been characterized in numerous tissues, including vessels. Recent studies demonstrate that interfering with either the distribution of chloride across the membrane or the ability of chloride channels to open markedly suppresses contractile responses of vascular smooth muscle to norepinephrine. Penile corpora own the unique ability to inflate and deflate itself. Based on the obvious similarities in the structural and pharmacological properties between cavernous myocytes and vascular smooth muscle cells, Clmight be expected to also play a role in the regulation of the cavernous smooth muscle tone, and thus, the erectile function. Our previous studies showed that the CLC-3 chloride channel proteins could be detected in rat corpus cavernous smooth muscle tissue and cells via western blot. This result means more extensive molecular studies of chloride channels in erectile tissue may be necessary to distinguishing its possible physiological role. We therefore conducted a strategy involving: 1. Immunohistochemistry Detection of the distribution of CLC-3 chloride channel protein in rat corporal tissues and culture cells. 2. Cloning and preparation of probes The probes are synthesized by the primers by PCR amplification. 3.. Northern blot analysis Detection of CLC-3 chloride channel gene expression in rat corporal tissues and culture cells. The results revealed that the method of immunohistochemistry stain could identify the distribution of CLC-3 chloride channel protein in could be detected on rat corporal tissues. The probes could be synthesized by the primers by PCR amplification.application/pdf57782 bytesapplication/pdfzh-TW國立臺灣大學醫學院泌尿科氯離子通道海綿體平滑肌ClC-3Chloride channelCorpus cavernous smooth muscleotherhttp://ntur.lib.ntu.edu.tw/bitstream/246246/25517/1/922314B002333.pdf