ROCK1 IN DIABETIC NEPHROPATHY

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 法舒地尔抑制Rho激酶(ROCK)活性改善糖尿病进展 肾病(DN)。然而，法舒地尔不是选择性的，也不能区分 ROCK-1和ROCK-2是ROCK的两种不同异构体，在糖尿病肾病中的致病作用。因此，在 在目前的研究中，我们将使用遗传学的方法。我们在FVB上产生了ROCK1-/-小鼠 背景资料。这些小鼠的表型是正常的。 具体目的：评价ROCK1在线粒体断裂中的作用。 我们观察到ROCK1/-糖尿病小鼠表现出显著的减少 线粒体碎裂与野生型糖尿病小鼠的比较。因此，我们正在 评估ROCK1在肾小球线粒体碎裂中的作用 具有2D和3D EM的足细胞。 为此，不会给糖尿病和糖尿病小鼠灌流，并固定它们的肾脏和 为染色、塑料包埋和切片做好准备。 在先前研究的基础上，我们计划从序列中重建线粒体结构 用于确认线粒体碎裂的代表性区域的切片 糖尿病小鼠的足细胞。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Inhibition of Rho kinase (ROCK) activity by fasudil ameliorates progression of diabetic nephropathy (DN). However, fasudil is not selective, and can not distinguish the pathogenic roles of ROCK-1 vs. ROCK-2, two different isomers of ROCK, in DN. Therefore in the current study, we will use a genetic approach. We generated ROCK1-/- mice on FVB background. These mice are phenotypically normal. Specific Aim: To evaluate the role of ROCK1 on mitochondrial fragmentation. We observed that ROCK1-/- diabetic mice demonstrated a significant reduction in mitochondrial fragmentation compared to wild type diabetic mice. Therefore, we are planning to evaluate the role of ROCK1 on mitochondrial fragmentation in glomerular podocytes with 2D and 3D EM. To this end, none diabetic and diabetic mice will be perfused, and their kidneys fixed and prepared for stained, plastic embedding and sectioning. Based on a previous study, we plan to reconstruct mitochondrial structure from serial sections of a representative region to confirm the mitochondrial fragmentation in podocytes from diabetic mice.