Robert Hannah, Ferguson Lindsay, Reins Olivia, Greco Tiffany, Prins Mayumi L, Folkerts Michael
Department of Neurosurgery, Brain Injury Research Center, UCLA, David Geffen School of Medicine;
Department of Neurosurgery, Brain Injury Research Center, UCLA, David Geffen School of Medicine.
J Vis Exp. 2021 Aug 30(174). doi: 10.3791/62884.
The current methodology establishes a reproducible, standardized, and cost-effective approach to monitoring the estrous cycle of female Sprague Dawley (SD) adolescent rats. This study demonstrates the complexity of hormonal cycles and the broad spectrum of understanding required to construct a reliable and valid monitoring technique. Through an in-depth examination of principal experimental design and procedural elements, this description of the cycle and its fundamental principles provides a framework for further understanding and deconstructs misconceptions for future replication. Along with an outline of the sample collection process employing vaginal lavage, the procedure describes the mechanism of data categorization into the four-stage model of proestrus, estrus, metestrus, and diestrus. These stages are characterized by a new proposed approach, utilizing the 4 categorizing determinants of vaginal fluid condition, cell type(s) present, cell arrangement, and cell quantity at the time of collection. Variations of each stage, favorable and unfavorable samples, the distinction between cyclicity and acyclicity, and graphic depictions of the collected categorizing components are presented alongside effective interpretive and organizational practices of the data. Overall, these tools allow for the publication of quantifiable data ranges for the first time, leading to the standardization of categorization factors upon replication.
当前的方法建立了一种可重复、标准化且具有成本效益的方法来监测雌性斯普拉格-道利（SD）青春期大鼠的发情周期。本研究展示了激素周期的复杂性以及构建可靠且有效的监测技术所需的广泛理解。通过对主要实验设计和程序要素的深入研究，对该周期及其基本原理的描述提供了一个进一步理解的框架，并解构了未来重复实验时的误解。除了概述采用阴道灌洗的样本采集过程外，该程序还描述了将数据分类到发情前期、发情期、发情后期和间情期四阶段模型的机制。这些阶段的特征是采用一种新提出的方法，利用采集时阴道液状况、存在的细胞类型、细胞排列和细胞数量这4个分类决定因素。文中呈现了每个阶段的变化、有利和不利样本、周期性和非周期性的区别，以及所采集分类成分的图形描述，同时还给出了有效的数据解释和组织方法。总体而言，这些工具首次实现了可量化数据范围的公布，从而在重复实验时实现了分类因素的标准化。
