Cyst induction and growth in ADPKD

所要求的预算将用于生成数据，以最终确定我们的手稿，提出一个机械目标1，并处理审查人员的其他意见。具体而言，该R56将涵盖以下初步实验：1）使用Hprt-cre和UBC-cre小鼠的高重组MADM模型的生成和表征，2）Six 2-cre和Ksp-cre MADM模型的表征，包括用近端、Henle环、远端和集合小管标记物的抗体染色以表征肾小管上皮细胞的表达。 识别囊肿，量化囊肿数量和大小，以及囊肿中细胞随时间的增殖/衰老;和3）我们将在用TAT-cre诱导的发育中的肾脏上进行突变和对照细胞的活体成像，并表征在囊肿发育开始时突变细胞的增殖和细胞迁移。

The requested budget will be used to generate data to finalize our manuscripts, to propose a mechanistic aim 1 and to address additional comments from the reviewers. Specifically, the following preliminary experiments will be covered by this R56: 1) Generation and characterization of high-recombination MADM models using Hprt-cre and UBC-cre mice, 2) Characterization of the Six2-cre and Ksp-cre MADM model, including staining with antibodies for proximal, Loop of Henle, distal and collecting tubule markers to characterize the tubular identify of the cysts, quantification of cyst number and size, and proliferation/senescence of cells in the cysts over time; and 3) We will perform live-imaging of mutant and control cells on developing kidneys induced with TAT-cre and characterize proliferation and cell migration of mutant cells at the inception of cyst development.