GAP JUNCTIONS & ION CHANNELS IN CORPUS CAVERNOSUM

缝隙连接

• 批准号: 2713378
• 负责人: George Joseph Christ
• 金额: $ 23.78万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-06-01 至 1999-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2713378
• 关键词: action potentials; calcium channel; electrophysiology; fluorescent dye /probe; gap junctions; gene expression; human subject; immunoelectron microscopy; immunofluorescence technique; impotence; membrane channels; messenger RNA; muscle cells; muscle contraction; muscle relaxation; muscle tone; northern blottings; penis disorder; penis erection; potassium channel; second messengers; smooth muscle; tissue /cell culture; voltage /patch clamp; western blottings

Erectile dysfunction is a major health problem that has a dramatic impact on the quality of life of many men and their sexual partners. Moreover, erectile dysfunction represents a spectrum of disease, ranging from partial to complete impotence, and affecting an estimated 18-30 million American men greater than or equal to 40 years of age. Medical treatment of this prevalent disease resulted in 400,000 outpatient visits and 30,000 hospital admissions, at total cost of $146 million in 1985 alone. Incomplete corporal smooth muscle relaxation is now widely recognized as a significant etiologic factor in a large proportion of impotent men. The general consensus is that incomplete corporal smooth muscle relaxation severely compromises trapping of blood in the corporal sinuses (because of incomplete closure of the venous outflow), resulting in a lack of rigidity. This condition is commonly referred to as corporal veno- occlusive erectile dysfunction. In vitro studies have documented that isolated human corporal tissue strips and cultured corporal smooth muscle cells provide a valid model for studying at least some aspects of the modulation of corporal smooth muscle tone in vivo. Observations both in vitro and in vivo demonstrated that intercellular communication through gap junctions, and current flow through membrane ion channels (i.e., namely Ca & K channels) are important modulators of corporal smooth muscle tone, and therefore, of erectile capacity. It seems that regardless of the diversity of causes of erectile dysfunction related to incomplete corporal smooth muscle relaxation, their effects might still be explained via their direct or indirect impact on gap junctions, K channels or Ca channels. Thus, in many ways, the improved understanding, diagnosis and treatment of erectile dysfunction is largely dependent on more detailed knowledge of how physiologically relevant drugs modulate these primary effectors of corporal smooth muscle tone. To directly address this issue we shall: l: a) Conduct electrophysiological studies on enzymatically dispersed and cultured corporal smooth muscle cells to evaluate the role of intercellular current flow in propagating and amplifying signals in the corpora. b) Microinject cultured and enzymatically dissociated cells as well as isolated tissue strips with gap junction permeant dyes and second messenger molecules to assess the role of metabolic coupling in propagating and amplifying signals in the corpora. c) Conduct molecular biological and immunocytochemical studies to characterize gap junctions between smooth muscle cells in situ and in culture. 2) Use patch clamp techniques to characterize the K and Ca channels in enzymatically dispersed and cultured corporal smooth muscle cells. 3) Assess action potential generation in vitro, using the sucrose gap technique.

勃起功能障碍是一个具有重大影响的主要健康问题。 许多男性和他们的性伴侣的生活质量。此外， 勃起功能障碍代表一系列疾病，范围从 偏向完全性阳萎，估计有1800-3000万人受到影响 年龄大于或等于40岁的美国男子。医疗 这一流行疾病导致400,000人次门诊和30,000人次 入院费用，单在一九八五年便达1.46亿元。 不完全的躯体平滑肌松弛现在被广泛认为是 这是很大一部分男性阳萎的重要病因。这个 普遍的共识是不完全的躯体平滑肌松弛 严重损害了下体鼻窦的血液滞留(因为 静脉流出的不完全闭合)，导致缺乏 僵硬。这种情况通常被称为下士维诺- 闭塞性勃起功能障碍。体外研究已经证明 人离体体组织条和体外培养的体表平滑肌 细胞至少为研究生物多样性的某些方面提供了有效的模型。 在体体肌张力的调节。观察到的数据 体外和体内研究表明，细胞间的通讯通过 缝隙结和流过膜离子通道的电流(即， 即钙钾通道)是体肌的重要调节剂 基调，因此，勃起能力。看起来，不管 与不完全躯体相关的勃起功能障碍原因的多样性 平滑肌松弛，它们的作用仍然可以通过它们的 直接或间接影响缝隙连接、钾通道或钙通道。 因此，在许多方面，对慢性阻塞性肺疾病的认识、诊断和治疗的改进 勃起功能障碍在很大程度上依赖于对 生理相关药物是如何调节这些主要效应器的 下士肌肉肌肉强直。为了直接解决这个问题，我们将：L： A)对酶分散的和 体外培养大鼠海绵体平滑肌细胞的实验研究 胞间电流在细胞内传播和放大信号 语料库。B)显微注射培养的和酶解离的细胞 以及带有缝隙连接的隔离组织条意味着染料和第二 信使分子在评估代谢偶联中的作用 在语料库中传播和放大信号。C)进行分子 缝隙连接的生物学和免疫细胞化学研究 在原位和培养中的平滑肌细胞之间。2)使用膜片钳 钾、钙通道的酶学表征技术 分散培养体部平滑肌细胞。3)评估行动 利用蔗糖间隙技术，在体外产生潜在的后代。

项目成果

K(ATP) channel currents regulate membrane potential in freshly isolated human and rat bladder smooth muscle cells.

K(ATP) 通道电流调节新鲜分离的人和大鼠膀胱平滑肌细胞的膜电位。

• DOI: 10.1016/s0090-4295(01)01039-1
• 发表时间: 2001
• 期刊: Urology
• 影响因子: 2.1
• 作者: Wang,HZ;Christ,GJ
• 通讯作者: Christ,GJ

Myocardial expression of endothelin-1 in murine Trypanosoma cruzi infection.

小鼠克氏锥虫感染中内皮素-1 的心肌表达。

• DOI: 10.1016/s1054-8807(00)00045-4
• 发表时间: 2000
• 期刊: Cardiovascular pathology : the official journal of the Society for Cardiovascular Pathology
• 作者: Petkova,SB;Tanowitz,HB;Magazine,HI;Factor,SM;Chan,J;Pestell,RG;Bouzahzah,B;Douglas,SA;Shtutin,V;Morris,SA;Tsang,E;Weiss,LM;Christ,GJ;Wittner,M;Huang,H
• 通讯作者: Huang,H

K channels as molecular targets for the treatment of erectile dysfunction.

K 通道作为治疗勃起功能障碍的分子靶点。

• 发表时间: 2002
• 期刊: Journal of andrology.
• 作者: Christ,GeorgeJ

Gap junction channel activity in short-term cultured human detrusor myocyte cell pairs: gating and unitary conductances.

短期培养的人逼尿肌细胞对中的间隙连接通道活性：门控和单一电导。

• DOI: 10.1152/ajpcell.00027.2006
• 发表时间: 2006
• 期刊: American journal of physiology. Cell physiology
• 作者: Wang,H-Z;Brink,PeterR;Christ,GeorgeJ
• 通讯作者: Christ,GeorgeJ

Trypanosoma cruzi infection (Chagas' disease) of mice causes activation of the mitogen-activated protein kinase cascade and expression of endothelin-1 in the myocardium.

小鼠的克氏锥虫感染（恰加斯病）会导致心肌细胞中有丝分裂原激活蛋白激酶级联的激活和内皮素-1 的表达。

• DOI: 10.1097/00005344-200036051-00046
• 发表时间: 2000
• 期刊: Journal of cardiovascular pharmacology
• 影响因子: 3
• 作者: Huang,H;Petkova,SB;Pestell,RG;Bouzahzah,B;Chan,J;Magazine,H;Weiss,LM;Christ,GJ;Lisanti,MP;Douglas,SA;Shtutin,V;Halonen,SK;Wittner,M;Tanowitz,HB
• 通讯作者: Tanowitz,HB