Department of Anatomy, Water and Salt Research Center, Aarhus University, DK-8000 Aarhus, Denmark.
Adv Exp Med Biol. 2011;704:277-86. doi: 10.1007/978-94-007-0265-3_15.
The transient receptor potential vanilloid 5 (TRPV5) is a Ca(2+) channel, which is expressed in renal late distal convoluted tubules (DCT2s) and connecting tubules (CNTs). These tubules play a major role in hormone controlled renal Ca(2+) reabsorption, and thereby in body Ca(2+) homeostasis, as well as urinary excretion of other electrolytes, including Na(+) and K(+). DCT2 and CNT are difficult to distinguish from the surrounding structures and thereby to study by direct functional methods. We developed a transgenic mouse model expressing enhanced green fluorescent protein (EGFP) driven by the TRPV5 promoter to identify these specific tubules. Expression of EGFP in the DCT2 and CNT allows the isolation of pure DCT2 and CNT populations for proteomic and physiological analyses. The TRPV5 promoter is also useful for generating conditional knockout mouse models in a cell-specific manner. TRPV5 promoter driven Cre recombinase expression will be useful for inducing DCT2 and CNT specific gene silencing of various channels, pumps, carriers, and receptors. In this chapter, we describe the strategy for developing transgenic mouse lines involving the TRPV5 promoter, provide a description of extensive validation of these mouse lines, and discuss possible uses and limitations.
瞬时受体电位香草酸 5 型通道（TRPV5）是一种钙通道，表达于肾脏远曲小管末段（DCT2s）和连接小管（CNTs）。这些小管在激素控制的肾脏钙重吸收中起主要作用，从而在体内钙稳态以及其他电解质（包括 Na+和 K+）的尿排泄中起作用。DCT2 和 CNT 难以与周围结构区分，因此难以通过直接功能方法进行研究。我们开发了一种表达增强型绿色荧光蛋白（EGFP）的转基因小鼠模型，该模型由 TRPV5 启动子驱动，用于鉴定这些特定的小管。DCT2 和 CNT 中 EGFP 的表达允许分离纯 DCT2 和 CNT 群体进行蛋白质组学和生理学分析。TRPV5 启动子也可用于以细胞特异性方式生成条件性敲除小鼠模型。TRPV5 启动子驱动的 Cre 重组酶表达将有助于诱导 DCT2 和 CNT 特定的各种通道、泵、载体和受体基因沉默。在本章中，我们描述了涉及 TRPV5 启动子的转基因小鼠系的开发策略，提供了对这些小鼠系的广泛验证的描述，并讨论了可能的用途和局限性。
