Department of Comparative Biosciences, University of Wisconsin-Madison, Madison, WI, USA.
University of Wisconsin-Madison/UMASS Boston/UT Southwestern George M. O'Brien Center for Benign Urologic Research, Madison, WI, USA.
Physiol Rep. 2021 Aug;9(15):e14964. doi: 10.14814/phy2.14964.
The void spot assay (VSA) is a cost-effective method for evaluating and quantifying mouse urinary voiding phenotypes. The VSA has been used to differentiate voiding behaviors between experimental groups, but not as a diagnostic assay. To build toward this goal, we used the VSA to define voiding patterns of male mice with diabetic diuresis (BTBR.Cg-Lep /WiscJ mice), irritative urinary dysfunction (E. coli UTI89 urinary tract infection), and obstructive urinary dysfunction (testosterone and estradiol slow-release implants) compared to their respective controls. Many studies compare individual VSA endpoints (urine spot size, quantity, or distribution) between experimental groups. Here, we consider all endpoints collectively to establish VSA phenomes of mice with three different etiologies of voiding dysfunction. We created an approach called normalized endpoint work through (NEW) to normalize VSA outputs to control mice, and then applied principal components analysis and hierarchical clustering to 12 equally weighted, normalized, scaled, and zero-centered VSA outcomes collected from each mouse (the VSA phenome). This approach accurately classifies mice based on voiding dysfunction etiology. We used principal components analysis and hierarchical clustering to show that some aged mice (>24 m old) develop an obstructive or a diabetic diuresis VSA phenotype while others develop a unique phenotype that does not cluster with that of diabetic, infected, or obstructed mice. These findings support use of the VSA to identify specific urinary phenotypes in mice and the continued use of aged mice as they develop urinary dysfunction representative of the various etiologies of LUTS in men.
空位分析（VSA）是一种评估和量化小鼠排尿表型的经济有效的方法。VSA 已被用于区分实验组之间的排尿行为，但不作为诊断分析。为了实现这一目标，我们使用 VSA 来定义具有糖尿病利尿（BTBR.Cg-Lep/WiscJ 小鼠）、刺激性尿功能障碍（大肠杆菌 UTI89 尿路感染）和阻塞性尿功能障碍（睾酮和雌二醇缓释植入物）的雄性小鼠的排尿模式与各自的对照相比。许多研究比较实验组之间的单个 VSA 终点（尿斑大小、数量或分布）。在这里，我们综合考虑所有终点来建立具有三种不同排尿功能障碍病因的小鼠的 VSA 表型。我们创建了一种称为归一化终点工作的方法（NEW），将 VSA 输出归一化为对照小鼠，然后应用主成分分析和层次聚类对从每只小鼠收集的 12 个同等权重、归一化、缩放和零中心化的 VSA 结果（VSA 表型）进行分析。这种方法可以根据排尿功能障碍的病因准确地对小鼠进行分类。我们使用主成分分析和层次聚类来表明，一些老年小鼠（>24 个月）会发展出阻塞性或糖尿病利尿 VSA 表型，而另一些则会发展出一种独特的表型，与糖尿病、感染或阻塞性小鼠的表型不聚类。这些发现支持使用 VSA 来识别小鼠的特定尿表型，并继续使用老年小鼠，因为它们会发展出代表男性下尿路症状（LUTS）各种病因的尿功能障碍。
