TENDON CELLS:INTERACTIONS & RESPONSES TO STRESS IN VITRO

描述(改编自申请者摘要)：细胞间 沟通对于大多数器官的体内平衡功能是必不可少的， 协调发育过程中的事件，并调整心脏收缩的速度， 肺、肠道、勃起组织和子宫。来自校长的已发布报告 研究小组现在指出，细胞活动的协调 通过缝隙连接在屈肌腱对机械负荷的反应中是至关重要的 还有伤人。运动对保持健康至关重要：正常的运动是 积极，固定是消极的，过度活动是破坏性的。在……里面 医学上，损伤后的运动疗法是恢复活动范围必不可少的。 申请者假设细胞活动通过GAP进行协调 在机械负荷和肌腱损伤后，关节是至关重要的。如果 缝隙连接被阻断，细胞协调将被阻止。以特定的目标 1，他们将测试缝隙连接在通过Ca~(2+)、GAP 与GFP-Cx43构建的连接易位以及DNA和胶原 体外机械加载和损伤后细胞的合成 缝隙连接堵塞。负荷将包括急性方案和适应性方案。 在特定的目标2中，他们将测试PKC调制对缝隙连接的影响 与化合物的表达和功能，如三磷酸腺苷和去甲肾上腺素，它们 调节缝隙连接信号和表达。在具体目标3和4中，他们 将进行类似于AIMS 1和2的实验，但在整个肌腱中， 以响应长达5天的适应性加载。连接蛋白43是一种机械性的 负荷敏感基因。肌腱细胞如何转导和反应的知识 机械载荷和创伤信号将影响我们对 患者在受伤后对运动疗法有反应。了解运动如何作用于 机制水平可能导致药物策略上调细胞 有无运动的老年人和残疾人的除法和矩阵表达 病人。

DESCRIPTION (Adapted from the Applicant's Abstract): Intercellular communication is essential for homeostatic function of most organs, to coordinate events during development, and to pace contractions in the heart, lungs, gut, erectile tissue and uterus. Published reports from the principal investigator's group now indicate that coordination of cellular activities through gap junctions is vital in flexor tendons in response to mechanical load and wounding. Exercise is critical to health maintenance: normal activity is positive, immobilization is negative and overactivity is destructive. In medicine, motion therapy post-injury is essential to recover range of motion. The applicants hypothesize that coordination of cellular activities through gap junctions is vital in response to mechanical load and after tendon injury. If gap junctions are blocked, cell coordination will be prevented. In Specific Aim 1, they will test the importance of gap junctions in signaling via Ca2+, gap junction translocation with a GFP-cx43 construct, and DNA and collagen synthesis in cells that have been mechanically loaded and wounded in vitro +/- gap junction blockage. Loading will include both acute and adaptive regimens. In Specific Aim 2, they will test the effect of PKC modulation on gap junction expression and function with compounds, such as ATP and norepinephrine, which regulate gap junction signaling and expression. In Specific Aims 3 and 4, they will conduct similar experiments to those in Aims 1 and 2, but in whole tendon, in response to adaptive loading for up to 5 days. Connexin 43 is a mechanical load-sensitive gene. Knowledge of how tendon cells transduce and respond to mechanical load and wound signals will have impact on our understanding of how patients respond to motion therapy post-injury. Knowledge of how motion acts at the mechanistic level may lead to drug strategies for upregulating cell division and matrix expression with or without motion in aged or disabled patients.