我们要感谢马克·伦纳德·卡尔顿身临其境的媒体工作室视频制作和编辑博士和马丁​​·贝特朗·卡尔顿身临其境的媒体工作室/神经再生实验室的可视化模型援助的专家技术援助。作者非常感谢玛丽莲Keaney博士和她的专职工作人员，在大学渥太华动物保护和兽医服务的专业意见。这项工作是由加拿大卫生研究院（CIHR研究所，澳门币62826）SALB，CIHR老龄研究所和战略的培训计划，在卫生研究/ CIHR培训的神经退行性脂类组学计划（TGF 96121）SALB和SF，加拿大基金会创新，安大略省创新信托SF，SF和Autodesk研究以SF。 ACM收到CIHR Banting和Best博士奖。 NV获得一个专业奖学金后，从人口老龄化和的CIHR培训计划在神经退行性脂类组学研究所。

1。准备试剂<ol><li>对于无菌阴道灌洗，高压灭菌双蒸水（DDH 2 O），并存储在一个密封的容器中，室温，直到需要。 </li><li>对于细胞学评估，结晶紫粉0.1克至100毫升DDH 2 O拌匀。结晶紫染色（0.1％）可被存储在一个密封的容器中，在室温下，直到需要。 </li></ol> 2。收集阴道细胞（阴道灌洗） <ol><li>胶乳灯泡放置在无菌的200μl的尖端的端部和绘制最多约100微升无菌DDH 2 O的前端中使用的灰度作为准则的体积。 </li><li>鼠标抬起她的笼子笼斗（盖），并把她与她的后腿/后端向您。 </li><li>紧紧抓住尾巴和提升的后端。鼠标现在只有她的前爪抓料斗。在这一点上，鼠标可能会排尿。如果是这样的，等到排尿站。如果有尿留在阴道的入口，你可能想使用一个单独的尖端（ 即 ，不是你的样品采集头）冲洗开幕多余的DDH 2 O。 </li><li>将在开幕式的阴道，注意不要穿透孔，阴道和宫颈的刺激能诱发假孕大鼠1,2的DDH 2 O填充尖。最近的报告表明，老鼠是不容易受到这种影响仍然应注意的侵袭程度，以尽量减少重复分析3。 </li><li>轻轻地踏下灯泡，驱逐一季度的一半量的水（约25-50微升）在开幕阴道管。无尖插入液体会自发地吸进运河。缓慢地释放在灯泡上的压力。流体将撤回到尖。避免释放压力的速度太快，以防止误吸液到灯泡。过滤的尖头可能是有用的用于此目的。 </li><li>重复前面的步骤使用相同的尖端，灯泡，和流体的4-5倍，在一个单一的样品，以获得足够数量的细胞。 </li><li>流体放置在玻璃片上，并允许涂抹标本在室温下完全干燥。发情涂片干燥后，可立即染色或存储，并在以后的日子染色。 </li></ol> 3。细胞学染色结晶紫* 4 <ol><li>将在科普林氏jar文件（或其他可比较的染色容器）中含有的结晶紫染色1分钟干滑动。 </li><li>删除科普林氏的第二个罐子DDH 2 O洗净幻灯片DDH 2 O为1分钟。重复以上步骤。 </li><li>删除多余的DDH 2 O幻灯片中一个轻型的组织，雨刮器的边缘，避免接触涂片染色。 </li><li>移液器约15微升的顶部的涂片和盖的甘油打滑。或者，其他组织学安装试剂可以利用，以获得一个更永久的，非扩散染色。 </li></ol> *这里描述的染色方法是最简单的程序，可以在任何实验室进行。其他的方法可以提供更多的细节。例如，采用巴氏染色法，有核上皮细胞的成熟度来区分不太成熟的细胞染色的粉红色或橙染色的绿松石和更成熟的细胞。这些差异可以被用来举办提前或延迟发情前期。 4。阴道细胞学检查<ol><li>检查涂片，光镜下观察，以确定细胞类型。结晶紫从细胞中扩散，随着时间的推移使用甘油时，盖片染色后镜检应立即做。在分析时，应采取显微照片记录细胞学检查。 </li><li>通过检查的entir开始Ë涂抹在较低的放大倍率。选择有代表性的区域并移动到一个更高的放大倍率。您将看到角化鳞状上皮细胞，白细胞，和/或有核上皮细胞（ 具有代表性的结果 ， 图1A-C）。细胞存在的比例将允许你确定你的鼠标动情期，在取样时（ 具有代表性的结果 ， 图1D-G）和她的激素水平（ 讨论 ， 图2）。 </li></ol> 5。代表性的成果 细胞学检查：三种主要的类型的细胞中可检测到阴道涂片样品：（1）有核上皮细胞（ 图1A），（2）角化鳞状上皮细胞（ 图1B），和（3）白细胞（ 图1C）。有核上皮细胞胞浆淡染，较暗的彩色质膜，以及一个椭圆形的核（ <strONG&gt;图1A）。角化鳞状上皮细胞是均匀地染色，更多边形的形状比他们的有核上皮前辈，和没有细胞核（ 图1B）。多形核白细胞，上皮细胞可区别于其形状不规则，深染多态性核，和小尺寸（ 图1C中 ，黑色箭头所示）。如果尿污染本在涂片，尿酸结晶很容易检测到由异种的任何预期类型的细胞（ 图3）的结晶结构体。如果发生这种情况，并不起眼的检测主要的细胞类型，涂抹应该被丢弃，而不是用于举办目的。 分段：涂片中观察到的细胞类型的相对比率上的一天的样品收集（ 图1D-G），可以用来识别你的鼠标的动情周期的阶段。在发情前期，细胞几乎完全集群的圆形，形成有核上皮细胞（ 图1D，代表细胞由白色箭头指示）。在发情期，细胞主要是角化鳞状上皮细胞，存在在密密麻麻的集群（ 图1E，代表细胞的箭头指示）。在发情后期，小深染白细胞占主导地位（ 图1F，代表电池用黑色箭头表示）。角化鳞状上皮细胞中可以观察到，往往在片段，（ 图1F，代表性的细胞用黑箭头表示）。在动情，罕见角化鳞状上皮细胞仍可能存在（ 图1G，代表性的细胞用黑色箭头表示），白细胞仍然占主导地位（ 图1G，代表性的细胞用黑色箭头表示）。发情后期可区别于发情间期核上皮细胞的外观在发情间期（ <stron<html> G&gt;图1G，代表细胞的白色箭头指示）。 <img src="/files/ftp_upload/4389/4389fig1.jpg" alt="图1" /> 图1。阴道涂片的细胞学评估可以用于识别动情期，三个主要类型的细胞中检测到阴道涂片样品的：（A）的有核上皮细胞，（B）的角化鳞状上皮细胞，和（C）的白细胞。涂片这些类型的细胞中存在的比率，可以使用（D）的动情前期，（E）的发情期，（F）的发情后期，或（G）的发情间期中描述的有代表性的结果 ，以确定小鼠。黑色箭头代表角化鳞状上皮细胞E，F和G点。黑色箭头C，F和G点代表leykocytes。在D和G的亮点代表的白色箭头核上皮细胞。 文件，“ALT =”图2“/&gt; 图2。阴道涂片细胞学检查反映潜在的内分泌活动。还提供了详细讨论。 <a href="https://www.jove.com/files/ftp_upload/4389/4389fig2large.jpg" target="_blank">点击此处查看大图</a> 。 <img src="/files/ftp_upload/4389/4389fig3.jpg" alt="图3" /> 图3。尿酸结晶可能会出现下面的结晶紫染色尿污染的样本 （A）的晶体是透明的，可以是各种尺寸（箭头和盒装（B）中放大的区域）。没有在这一领域存在的细胞。如果字段内的污染进行细胞学染色尿酸结晶存在，它可能难以准确地确定细胞类型，涂抹应该被丢弃。比例尺= 50微米。

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N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Estrous+cycle+modulates+ovarian+carcinoma+growth.">Estrous cycle modulates ovarian carcinoma growth.</a> Clinical cancer research : an official journal of the American Association for Cancer Research. 15, 2971-2978 (2009).</li><li>Jemiolo, B., Harvey, S., Novotny, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Promotion+of+the+Whitten+effect+in+female+mice+by+synthetic+analogs+of+male+urinary+constituents.">Promotion of the Whitten effect in female mice by synthetic analogs of male urinary constituents.</a> Proceedings of the National Academy of Sciences of the United States of America. 83, 4576-4579 (1986).</li></ol>

作者宣称没有利益冲突。所有的动物实验均严格按照渥太华大学动物保健委员会和加拿大动物保护协会的准则和规定。

细胞类型学的这些变化是潜在的内分泌活动的指标。动情前期阶段的动情周期对应的月经周期的5人卵泡阶段定义由排卵前增加循环中17-β-雌二醇的水平6，以及一个小的浪涌在催乳素7（ 图2，发情前期，左图）。的增加在17-β-雌二醇的间接刺激促性腺激素释放激素神经元的，反过来，激活响应在垂体前叶细胞释放促黄体激素和促卵泡激素的循环8,9（ 图2的下丘脑和隔垫，发情前期，左图）。取自动物在发情前期阴道分泌物涂片检查，细胞几乎完全是椭圆形的，有核上皮细胞（ 图1D，图2中 ，动情前期，右图）。高峰，卵泡刺激HORM1电平信号排卵进入发情期10,11。在发情期间，17-β-雌二醇水平下降和催乳素水平峰值6,7（ 图2，发情期，左侧面板）。阴道脱落细胞涂片的特点是几乎是唯一的检测不规则形角化鳞状上皮细胞通常在团块（ 图2，图1E，发情期，右图）。进入发情后期恰逢连续上涨的孕激素水平6，对应于开始的12人黄体期（ 图2，动情后期，左侧面板）。由于孕激素水平开始上升，在17-β-雌二醇水平在黄体激活6,13,14（ 图2，动情后期，左侧面板），有一个小高潮。在这个阶段，阴道分泌物涂片检查中存在的细胞类型是零散的，角化上皮细胞和较深的染色白细胞（FIGURË1F， 图2，动情后期，右图）。最后，发生进入发情间期的小鼠和循环黄体酮水平峰值6，对应于人类晚黄体期12。回归的黄体导致随后急剧下降，孕激素水平15,16（ 图2，间情期，左侧面板）。白细胞中占主导地位涂片在动情。角化上皮细胞的频率降低和成核的上皮细胞开始被检测到之前，过渡到动情前期（ 图1G，图2，求偶期，右面板）。 总之，这个简单的，日常的协议可用于生育状况没有改变，如果采取以下的预防措施，估计每天荷尔蒙的波动，并建立实验小鼠的动情期。抽样应在不超过每日一次使用的非侵入性的协议递减ribed相比，阴道，愿望和搅拌，反复渗透。这可能会导致阴道刺激导致的炎症反应17白细胞和其它类型的细胞中产生的，是本涂片可能混淆细胞学评估。此外，即使在殖民地安置女性，这是正常的扩展发情间期和发情阶段，在不同的鼠标，以及诱导休情期18和识别的解释是非常有用的激素影响生殖，性别，疾病的研究。随着年龄的增长和住房群内差异（个人或集团-房屋）的女性7,19-21还引入了变异周期长。女性女性，只有殖民地坐落在停止循环，并进入虽然循环可以重新设置曝光网箱预处理引起男性尿液循环24,25长期动情18,22,23的状态。因此，要建立个别升周期长度的给定的鼠标，建议每日执行这里描述的非侵入性的评估，小心使用，直到观察到两个完整的周期。

<table border="1"><tbody><tr><td> 名称 </td><td> 公司 </td><td> 目录＃ </td></tr><tr><td>无菌200微升吸头</td><td> Diamed </td><td> E340901 </td></tr><tr><td>乳胶灯泡（1毫升） </td><td>费舍尔</td><td> 03-488-21 </td></tr><tr><td>显微镜幻灯片</td><td>费舍尔</td><td> 12-550-15 </td></tr><tr><td>结晶紫染色（25克） </td><td>费舍尔</td><td> C581-25 </td></tr><tr><td>轻型组织雨刷的</td><td> VWR </td><td> 82003-820 </td></tr><tr><td>甘油</td><td>费舍尔</td><td> BP229-1 </td></tr><tr><td>显微镜盖玻片（22x30） </td><td>费舍尔</td><td> 12-544A </td></tr></tbody></table>

performing vaginal lavage, crystal violet staining, and vaginal cytological evaluation for mouse estrous cycle staging identification

在啮齿类动物的生殖状态评估是一种快速的方法不仅可用于生殖功能障碍的研究，但也需要生产新的小鼠模型的疾病和调查的激素调节组织变性（或再生）病理挑战。小鼠生殖或发情周期分为4个阶段：发情前期，发情期，发情后期和动情。定义卵巢激素17-β-雌二醇和孕酮的循环水平的波动，促性腺激素促黄体生成和促卵泡激素，黄体生成激素催乳激素信号通过这些生殖阶段过渡。小鼠阴道内的细胞类型学的变化反映了这些潜在的内分泌活动。每日评估有核上皮细胞，角化鳞状上皮细胞，和白细胞阴道涂片中存在的相对比例可以用于识别小鼠发情阶段。程度的侵袭，但是，受聘于收集这些样本可以改变生育状况和引起的炎症反应，可以混淆涂片细胞学检查评估。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小鼠的动情周期的阶段没有改变她的生殖周期。我们详细介绍了如何区分通过收集和分析阴道分泌物涂片检查的主要细胞类型学在发情周期的四个阶段，我们可以解释这些变化是如何与内分泌状态。

Watch this Scientific Journal Video about Performing Vaginal Lavage, Crystal Violet Staining, and Vaginal Cytological Evaluation for Mouse Estrous Cycle Staging Identification at JoVE.com

Performing Vaginal Lavage, Crystal Violet Staining, and Vaginal Cytological Evaluation for Mouse Estrous Cycle Staging Identification

在这里，我们介绍了如何识别阶段的小鼠的生殖发情前期，发情期，发情后期，或动情简单的，非侵入性的收集和阴道涂片细胞学评估。我们还介绍了如何阴道细胞学检查反映循环过渡，通过对小鼠的生殖周期的激素水平相关。

进行阴道灌洗，结晶紫染色法和阴道细胞学评价小鼠发情周期分期鉴定

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 ...

performing-vaginal-lavage-crystal-violet-staining-vaginal-cytological

Research

JoVE Journal

Biology

112.5K 次观看.  Neural Regeneration Laboratory and Ottawa Institute of Systems Biology. 在这里，我们介绍了如何识别阶段的小鼠的生殖发情前期，发情期，发情后期，或动情简单的，非侵入性的收集和阴道涂片细胞学评估。我们还介绍了如何阴道细胞学检查反映循环过渡，通过对小鼠的生殖周期的激素水平相关。

视频: Performing Vaginal Lavage, Crystal Violet Staining, and Vaginal Cytological Evaluation for Mouse Estrous Cycle Staging Identification

Protocol for Studying Extinction of Conditioned Fear in Naturally Cycling Female Rats

Extinction of conditioned fear has been extensively studied in male rodents. Recently, there have been an increasing number of studies indicating that neural mechanisms for certain behavioral tasks and response behaviors are different in females and males. Using females in research studies can represent a challenge because of the variation of gonadal hormones during their estrous cycle. This protocol describes well-established procedures that are useful in investigating the role of estrogen in fear extinction memory consolidation in female rats. Phase of the estrous cycle and exogenous estrogen administration prior to extinction training can influence extinction recall 24 hr later. The vaginal swabbing technique for estrous phase identification described here aids the examination and manipulation of naturally cycling gonadal hormones. The use of this basic rodent model may further delineate the mechanisms by which estrogen can modulate fear extinction memory in females.

Gonadal hormones such as estrogen modulate memory formation in a number of experimental paradigms including fear extinction memory. This protocol describes a set of methods for investigating the influence of gonadal hormones specifically during extinction in naturally cycling females, including estrous cycle monitoring and exogenous estrogen administration.

协议在骑自行车自然雌性大鼠学习条件性恐惧灭绝

Extinction of conditioned fear has been extensively studied in male rodents. Recently, there have been an increasing number of studies indicating that ...

科研

行为学

A Hyperandrogenic Mouse Model to Study Polycystic Ovary Syndrome

Hyperandrogenemia plays a critical role in reproductive and metabolic function in females and is the hallmark of polycystic ovary syndrome. Developing a lean PCOS-like mouse model that mimics women with PCOS is clinically meaningful. In this protocol, we describe such a model. By inserting a 4 mm length of DHT (dihydrotestosterone) crystal powder pellet (total length of pellet is 8 mm), and replacing it monthly, we are able to produce a PCOS-like mouse model with serum DHT levels 2 fold higher than mice not implanted with DHT (no-DHT). We observed reproductive and metabolic dysfunction without changing body weight and body composition. While exhibiting a high degree of infertility, a small subset of these PCOS-like female mice can get pregnant and their offspring show delayed puberty and increased testosterone as adults. This PCOS-like lean mouse model is a useful tool to study the pathophysiology of PCOS and the offspring from these PCOS-like dams.

We describe the development of a lean PCOS-like mouse model with&#160;dihydrotestosterone pellet to study the pathophysiology of PCOS and the offspring from these PCOS-like dams.

Hyperandrogenic 小鼠模型研究多囊卵巢综合征

Hyperandrogenemia plays a critical role in reproductive and metabolic function in females and is the hallmark of polycystic ovary syndrome. Developing a ...

发育生物学

Microdissection and Dissociation of the Murine Oviduct: Individual Segment Identification and Single Cell Isolation

Mouse model systems are unmatched for the analysis of disease processes because of their genetic manipulability and the low cost of experimental treatments. However, because of their small body size, some structures, such as the oviduct with a diameter of 200-400 &#956;m, have proven to be relatively difficult to study except by immunohistochemistry. Recently, immunohistochemical studies have uncovered more complex differences in oviduct segments than were previously recognized; thus, the oviduct is divided into four functional segments with different ratios of seven distinct epithelial cell types. The different embryological origins and ratios of the epithelial cell types likely make the four functional regions differentially susceptible to disease. For example, precursor lesions to serous intraepithelial carcinomas arise from the infundibulum in mouse models and from the corresponding fimbrial region in the human fallopian tube. The protocol described here details a method for microdissection to subdivide the oviduct in such a way to yield a sufficient amount and purity of RNA necessary for downstream analysis such as reverse transcription-quantitative PCR (RT-qPCR) and RNA sequencing (RNAseq). Also described is a mostly non-enzymatic tissue dissociation method appropriate for flow cytometry or single cell RNAseq analysis of fully differentiated oviductal cells. The methods described will facilitate further research utilizing the murine oviduct in the field of reproduction, fertility, cancer, and immunology.

A method for microdissection of the mouse oviduct that allows collection of the individual segments while maintaining RNA integrity is presented. In addition, non-enzymatic oviductal cell dissociation procedure is described. The methods are appropriate for subsequent gene and protein analysis of the functionally different oviductal segments and dissociated oviductal cells.

小鼠输卵管的显微解剖和解离:个体片段鉴定和单细胞分离

Mouse model systems are unmatched for the analysis of disease processes because of their genetic manipulability and the low cost of experimental ...

A Murine Model of Group B Streptococcus Vaginal Colonization

Streptococcus agalactiae (group B Streptococcus, GBS), is a Gram-positive, asymptomatic colonizer of the human gastrointestinal tract and vaginal tract of 10 - 30% of adults. In immune-compromised individuals, including neonates, pregnant women, and the elderly, GBS may switch to an invasive pathogen causing sepsis, arthritis, pneumonia, and meningitis. Because GBS is a leading bacterial pathogen of neonates, current prophylaxis is comprised of late gestation screening for GBS vaginal colonization and subsequent peripartum antibiotic treatment of GBS-positive mothers. Heavy GBS vaginal burden is a risk factor for both neonatal disease and colonization. Unfortunately, little is known about the host and bacterial factors that promote or permit GBS vaginal colonization. This protocol describes a technique for establishing persistent GBS vaginal colonization using a single &#946;-estradiol pre-treatment and daily sampling to determine bacterial load. It further details methods to administer additional therapies or reagents of interest and to collect vaginal lavage fluid and reproductive tract tissues. This mouse model will further the understanding of the GBS-host interaction within the vaginal environment, which will lead to potential therapeutic targets to control maternal vaginal colonization during pregnancy and to prevent transmission to the vulnerable newborn. It will also be of interest to increase our understanding of general bacterial-host interactions in the female vaginal tract.

A Murine Model of Group B Streptococcus Vaginal Colonization

The purpose of this protocol is to imitate human group B Streptococcus (GBS) vaginal colonization in a murine model. This method may be used to investigate host immune responses and bacterial factors contributing to GBS vaginal persistence, as well as to test therapeutic strategies.

B组的小鼠模型<em&gt;链球菌</em&gt;阴道定植

Streptococcus agalactiae (group B Streptococcus, GBS), is a Gram-positive, asymptomatic colonizer of the human gastrointestinal tract and vaginal tract of ...

免疫与感染

Determination of Reproductive Competence by Confirming Pubertal Onset and Performing a Fertility Assay in Mice and Rats

Assessment of reproductive competence is critical for understanding the impact of a treatment or genetic manipulation on the reproductive axis, also termed the hypothalamic-pituitary-gonadal axis. The reproductive axis is a key integrator of environmental and internal input adapting fertility to favorable conditions for reproduction. Prior to embarking upon a fertility study in mice and rats, sexual maturity is evaluated to exclude the possibility that the observed reproductive phenotypes are caused by delayed or absent pubertal onset. This protocol describes a non-invasive approach to assess pubertal onset in males through the determination of preputial separation, and in females through vaginal opening and first estrus. After the confirmation of the completion of puberty and the achievement of sexual maturity, a fertility study can be initiated. The procedure describes the optimal breeding conditions for mice and rats, how to set up a fertility study, and what parameters to evaluate and determine if the treatment or gene deletion has an impact on fertility.

Many treatments and genetic mutations impact the timing of sexual maturity and fertility. This protocol describes a non-invasive method to evaluate pubertal onset in mice and rats prior to setting up a fertility study in sexually mature animals.

在小鼠和大鼠中确认青春期发作和执行生育力测定生殖能力

Assessment of reproductive competence is critical for understanding the impact of a treatment or genetic manipulation on the reproductive axis, also ...

生物学

Ex Vivo Method for Assessing the Mouse Reproductive Tract Spontaneous Motility and a MATLAB-based Uterus Motion Tracking Algorithm for Data Analysis

Dysmenorrhea, or painful cramping, is the most common symptom associated with menses in females and its severity can hinder women&#39;s everyday lives. Here, we present an easy and inexpensive method that would be instrumental for testing new drugs decreasing uterine contractility. This method utilizes the unique ability of the entire mouse reproductive tract to exhibit spontaneous motility when maintained ex vivo in a Petri dish containing oxygenated Krebs buffer. This spontaneous motility resembles the wave-like myometrial activity of the human uterus, referred to as endometrial waves. To demonstrate the effectiveness of the method, we employed a well-known uterine relaxant drug, epinephrine. We demonstrate that the spontaneous motility of the entire mouse reproductive tract can be quickly and reversibly inhibited by 1 &#181;M epinephrine in this Petri dish model. Documenting the changes of uterine motility can be easily done using an ordinary smart phone or a sophisticated digital camera. We developed a MATLAB-based algorithm allowing motion tracking to quantify spontaneous uterine motility changes by measuring the rate of uterine horn movements. A major advantage of this ex vivo approach is that the reproductive tract remains intact throughout the entire experiment, preserving all intrinsic intrauterine cellular interactions. The major limitation of this approach is that up to 10-20% of uteri may exhibit no spontaneous motility. Thus far, this is the first quantitative ex vivo method for assessing spontaneous uterine motility in a Petri dish model.

Uterine contractions are important for the well-being of females. However, pathologically increased contractility may result in dysmenorrhea, especially in younger females. Here, we describe a simple ex vivo preparation allowing quick assessment of the efficacy of smooth muscle relaxants that may be used for treating dysmenorrhea.

用于评估小鼠生殖道自发性的方法和用于数据分析的基于 MATLAB 的 Uterus 运动跟踪算法

Dysmenorrhea, or painful cramping, is the most common symptom associated with menses in females and its severity can hinder women&#39;s everyday lives. ...

Rodent Estrous Cycle Monitoring Utilizing Vaginal Lavage: No Such Thing As a Normal Cycle

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,&#160;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.

This study details the crucial factors to consider in experimental designs involving female rats. In a larger sense, these data serve to decrease stigma and assist in the development of more inclusive diagnostic and intervention tools.

利用阴道灌洗的啮齿动物发情周期监测:没有正常周期这样的事情

The current methodology establishes a reproducible, standardized, and cost-effective approach to monitoring the estrous cycle of female Sprague Dawley ...

宫颈推拿诱导小鼠假孕用于辅助生殖

Inducing Pseudopregnancy in Female Mice Without the Need for Vasectomized Males Prior to Non-Surgical Embryo Transfer or Artificial Insemination

For successfully maintaining pregnancy with embryo transfer or artificial insemination,&#160;female recipient mice must be induced into a pseudopregnant state. Female mice are traditionally paired overnight with vasectomized males, and the following morning, the presence of a copulation plug is assessed. To increase the efficiency of producing pseudopregnant females, a cervical manipulation technique has been standardized to be used in combination with non-surgical embryo transfer or artificial insemination techniques in mice. The blunt end of a small plastic rod is inserted vaginally to contact the cervix and is vibrated for 30 s by contact with a trimmer. The procedure is quick and does not require anesthesia or analgesia. This technique increases the reliability and predictability of producing pseudopregnant females and entirely eliminates the requirement for vasectomized males. For CD1 mice, the efficiency of pseudopregnancy induction using cervical manipulation was 83% for females in estrus (N = 76) but only 38% of females in estrus were plugged by vasectomized males (N = 24). Artificial insemination in CD1 mice was performed by estrus synchronization with hormones, cervical manipulation, and the uterine transfer of sperm. Artificial insemination recipients receiving cervical manipulation (N = 76) had a pregnancy rate of 72% and an average litter size of 8.3 pups. This method can also be used to produce pseudopregnant females for non-surgical embryo transfer. Therefore, inducing pseudopregnancy by cervical manipulation is a convenient and efficient alternative to mating with a vasectomized male when performing non-surgical assisted reproduction techniques. Using cervical manipulation provides 3Rs (replacement, reduction, and refinement) benefits for assisted reproduction techniques by reducing the number of animals required and eliminating the necessity for surgically altered males.

作者聚焦: 在非手术胚胎移植或人工授精之前，无需对雄性进行输精管切除术的雄性小鼠进行假性妊娠  

The cervical manipulation method presented can induce pseudopregnancy in mice without the need for breeding females with vasectomized males. The induction of pseudopregnancy is required for the success of non-surgical embryo transfer and non-surgical artificial insemination, both of which are also presented.

在非手术胚胎移植或人工授精之前诱导雌性小鼠假妊娠，而无需输精管切除雄性

For successfully maintaining pregnancy with embryo transfer or artificial insemination,&#160;female recipient mice must be induced into a pseudopregnant ...

进行阴道灌洗，结晶紫染色法和阴道细胞学评价小鼠发情周期分期鉴定

摘要

探索更多视频

此视频中的章节

协议在骑自行车自然雌性大鼠学习条件性恐惧灭绝

Hyperandrogenic 小鼠模型研究多囊卵巢综合征

小鼠输卵管的显微解剖和解离:个体片段鉴定和单细胞分离

B组的小鼠模型

在小鼠和大鼠中确认青春期发作和执行生育力测定生殖能力

用于评估小鼠生殖道自发性的方法和用于数据分析的基于 MATLAB 的 Uterus 运动跟踪算法

利用阴道灌洗的啮齿动物发情周期监测:没有正常周期这样的事情

在非手术胚胎移植或人工授精之前诱导雌性小鼠假妊娠，而无需输精管切除雄性

在非手术胚胎移植或人工授精之前诱导雌性小鼠假妊娠，而无需输精管切除雄性

在非手术胚胎移植或人工授精之前诱导雌性小鼠假妊娠，而无需输精管切除雄性