ROCK1 IN DIABETIC NEPHROPATHY

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子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，法舒地尔没有选择性，不能区分 ROCK-1与ROCK-2（ROCK的两种不同异构体）在DN中的致病作用。因此在 在目前的研究中，我们将使用遗传学方法。我们在FVB上产生ROCK 1-/-小鼠， 背景这些小鼠的表型正常。 具体目的：评估ROCK 1在线粒体碎片化中的作用。 我们观察到ROCK 1-/-糖尿病小鼠表现出显著降低， 与野生型糖尿病小鼠相比，因此我们 计划评估ROCK 1在肾小球线粒体断裂中的作用， 用2D和3D EM观察足细胞。 为此，将灌注无糖尿病小鼠和糖尿病小鼠，并固定它们的肾脏， 准备染色、塑料包埋和切片。 基于以前的研究，我们计划从序列中重建线粒体结构， 代表性区域的切片，以确认 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.