Performing vaginal lavage, crystal violet staining, and vaginal cytological evaluation for mouse estrous cycle staging identification.
Performing vaginal lavage, crystal violet staining, and vaginal cytological evaluation for mouse estrous cycle staging identification.
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DOI：
10.3791/4389
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发表时间：
2012-09-15
期刊：
影响因子：
--
通讯作者：
Bennett SA
中科院分区：
文献类型：
--
作者：
McLean AC;Valenzuela N;Fai S;Bennett SA
一种快速评估啮齿类动物生殖状态的方法不仅对研究生殖功能障碍有用，而且对生产新的疾病小鼠模型和研究病理挑战后组织退化（或再生）的激素调节也很有必要。鼠的生殖（或发情）周期分为4个阶段：发情前期、发情期、退情期和退情期。确定卵巢类固醇17-β-雌二醇和黄体酮、促性腺激素、促黄体生成素和促卵泡激素、促黄体生成素催乳素信号在这些生殖阶段的循环水平波动。小鼠阴道内细胞类型的变化反映了这些潜在的内分泌事件。每日评估阴道涂片中有核上皮细胞、角化鳞状上皮细胞和白细胞的相对比例，可用于确定小鼠的发情期。然而，在收集这些样本时所采用的侵入性程度可以改变生殖状态并引起炎症反应，从而混淆涂片的细胞学评估。在这里，我们描述了一个简单的，非侵入性的协议，可以用来确定一个雌性老鼠的发情周期的阶段，而不改变她的生殖周期。我们详细介绍了如何通过收集和分析阴道涂片中的主要细胞类型来区分发情周期的四个阶段，并展示了这些变化如何与内分泌状态相关。
A rapid means of assessing reproductive status in rodents is useful not only in the study of reproductive dysfunction but is also required for the production of new mouse models of disease and investigations into the hormonal regulation of tissue degeneration (or regeneration) following pathological challenge. The murine reproductive (or estrous) cycle is divided into 4 stages: proestrus, estrus, metestrus, and diestrus. Defined fluctuations in circulating levels of the ovarian steroids 17-β-estradiol and progesterone, the gonadotropins luteinizing and follicle stimulating hormones, and the luteotropic hormone prolactin signal transition through these reproductive stages. Changes in cell typology within the murine vaginal canal reflect these underlying endocrine events. Daily assessment of the relative ratio of nucleated epithelial cells, cornified squamous epithelial cells, and leukocytes present in vaginal smears can be used to identify murine estrous stages. The degree of invasiveness, however, employed in collecting these samples can alter reproductive status and elicit an inflammatory response that can confound cytological assessment of smears. Here, we describe a simple, non-invasive protocol that can be used to determine the stage of the estrous cycle of a female mouse without altering her reproductive cycle. We detail how to differentiate between the four stages of the estrous cycle by collection and analysis of predominant cell typology in vaginal smears and we show how these changes can be interpreted with respect to endocrine status.
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影响因子：
3.6
作者：
FELICIO, LS;NELSON, JF;FINCH, CE
通讯作者：
FINCH, CE
DOI：
10.1196/annals.1429.018
发表时间：
2008-01-01
期刊：
MENSTRUAL CYCLE AND ADOLESCENT HEALTH
影响因子：
--
作者：
Hawkinsa, Shannon M.;Matzuk, Martin M.
通讯作者：
Matzuk, Martin M.
影响因子：
2.9
作者：
ADLER, NT;RESKO, JA;GOY, RW
通讯作者：
GOY, RW
影响因子：
4
作者：
WHITTEN, WK
通讯作者：
WHITTEN, WK
DOI：
10.1073/pnas.83.12.4576
发表时间：
1986-06-01
影响因子：
11.1
作者：
JEMIOLO, B;HARVEY, S;NOVOTNY, M
通讯作者：
NOVOTNY, M