Abstract:
The voiding spot assay (VSA) is increasingly being adopted as a standard method for assessing mouse urinary function. However, VSA outcomes are highly sensitive to housing environment and procedural parameters. Many variables exist among laboratories, including analytical software, type of daily housing cage, transportation, and the time of the day. Some of these variables, such as the time of VSA and analytical software, have been shown to result in inconsistency and incomparability of data. In this study, we evaluated whether the results of VSA can be compared across laboratories by minimizing these variables. We found that analytical tools between Fiji and MATLAB are in good agreement in the quantification of VSA parameters, especially primary voiding spot (PVS) parameters. Unexpectedly, we found that mice housed in different daily home cages did not alter voiding patterns in a standard VSA cage. Nonetheless, we still recommend acclimation when performing VSA in unfamiliar cages. Notably, mice are highly sensitive to transportation and the time in the morning versus afternoon, which can induce significant changes in voiding patterns. Therefore, a standardized period among laboratories and allowing 2-3 days of rest for mice acclimation after transportation are necessary for VSA. Finally, we performed VSA using identical procedural parameters in two laboratories from two geographical locations to compare the results of VSA and concluded that it is possible to generate limited comparable VSA data, such as PVS volume.
摘要:
排泄斑点测定法(VSA)越来越多地用作评估小鼠泌尿功能的标准方法。然而,VSA结果对住房环境和程序参数高度敏感。实验室之间存在许多变量,包括分析软件,日常住房笼子的类型,交通运输,和一天中的时间。其中一些变量,例如VSA和分析软件的时间,已被证明会导致数据的不一致和不可比较性。在这项研究中,我们通过最小化这些变量来评估VSA的结果是否可以跨实验室进行比较.我们发现斐济和MATLAB之间的分析工具在量化VSA参数方面非常吻合,特别是初级排尿点(PVS)参数。出乎意料的是,我们发现,在标准的VSA笼中,饲养在不同的日常家庭笼子中的小鼠不会改变排尿模式。尽管如此,我们仍然建议在不熟悉的笼子中进行VSA时适应。值得注意的是,老鼠对交通和早上与下午的时间高度敏感,这可以引起排尿模式的显著变化。因此,VSA需要实验室之间的标准化时间,并在运输后使小鼠适应2-3天的休息。最后,我们在两个地理位置的两个实验室使用相同的程序参数进行了VSA,以比较VSA的结果,并得出结论,可以生成有限的可比VSA数据,如PVS卷。