Revell Dustin Z, Yoder Bradley K
Department of Cell, Developmental, and Integrative Biology, University of Alabama at Birmingham, Birmingham, AL, United States.
Department of Cell, Developmental, and Integrative Biology, University of Alabama at Birmingham, Birmingham, AL, United States.
Methods Cell Biol. 2019;154:67-83. doi: 10.1016/bs.mcb.2019.04.012. Epub 2019 May 20.
The renal primary cilium is a small microtubule-based appendage thought to have mechano/chemosensory roles detecting changes in the fluid passing through the nephron. Mutations affecting cilium structure or function of ciliary-localized proteins result in a spectrum of diseases termed ciliopathies, with prevalent phenotypes such as the formation of renal cysts and fibrosis. While many studies have been conducted using fixed kidney sections or live imaging of cells in culture to investigate the cilium, examination in the context of a living murine kidney remains to be conducted. Previously, our lab generated the SSTR3 mouse to study cilium dynamics in vivo and found novel cilium behaviors that occurred following alteration of heart rate, blood pressure, and tubule flow. In this manuscript, we utilize multiple transgenic mouse models and an abdominal window imaging approach to observe primary cilia and tubule flow dynamics, immune cell movement, and renal Ca signaling as it occurs in real time within a live mouse kidney. We present this window method as an approach that can be used in combination with various fluorescently labeled transgenic mice to investigate renal physiology, pathology, and function in vivo in longitudinal studies for as long as 5weeks.
肾初级纤毛是一种基于微管的小附属物，被认为具有机械/化学传感作用，可检测流经肾单位的液体变化。影响纤毛结构或纤毛定位蛋白功能的突变会导致一系列称为纤毛病的疾病，其常见表型包括肾囊肿和纤维化的形成。虽然已经使用固定的肾脏切片或培养细胞的实时成像进行了许多研究来调查纤毛，但在活体小鼠肾脏的背景下进行检查仍有待开展。此前，我们实验室培育了SSTR3小鼠以研究体内纤毛动力学，并发现了心率、血压和肾小管流量改变后出现的新型纤毛行为。在本手稿中，我们利用多种转基因小鼠模型和腹部窗口成像方法来实时观察活体小鼠肾脏内的初级纤毛和肾小管流动动力学、免疫细胞运动以及肾脏钙信号传导。我们提出这种窗口方法可与各种荧光标记的转基因小鼠结合使用，以在长达5周的纵向研究中体内研究肾脏生理学、病理学和功能。
