Name: collection of post-mating semen from the female reproductive tract and measurement of semen liquefaction in mice
Uploaded: 2017-11-18T14:00:00.000+00:00
Duration: 12 min 6 s
Description: Watch this Scientific Journal Video about Collection of Post-mating Semen from the Female Reproductive Tract and Measurement of Semen Liquefaction in Mice at JoVE.com

这项工作得到了兽医学院 (立) 启动基金对 WW 的支持。作者感谢塞拉奥尔森 (立) 对这篇手稿的批判性阅读。

本研究中使用的所有动物和程序都是根据华盛顿州立大学 (立) 动物保育和使用委员会的指导方针, 并遵照立批准的动物议定书 #4702 和 #4735 处理的。
1. 小鼠
<ol><li>
使用标准 C57BL/6J 成年雄性小鼠或其他有兴趣的品系。在温度和湿度控制的环境下维护动物, 使用ad 随意获取水和食物。
	<ol>
<li>使用雄性饲养者, 从8周大到10月大不等。在实验中只使用经过验证的育种者。</li>
	</ol>
</li>
	<li>
使用具有组织选择性删除生殖道雌激素受体α (由Esr1基因编码) 的成年雌性小鼠在上皮细胞中10 (Wnt7a.在本研究中, Esr1f/f, 称为挖空或 "KO") 和控制窝 (Esr1f/f, 称为 "CTRL")。
	<ol>
<li>使用8周大的女性。女性的年龄根据兴趣实验而变化。</li>
	</ol>
</li>
	<li>确定在早上08:00h 的动情周期的阶段。只在周期的发情阶段使用雌性。有关阴道涂抹和细胞学分析的详细方法, 请参阅以前发布的过程11。</li>
</ol>2. 交配和评估交配插头
<ol><li>在发情的那天 (16:00h), 把雌性放在雄性笼子里。</li>
	<li>第二天早上 (08:00h), 把雌鼠和雄性老鼠分开。</li>
	<li>使用以前发布的方法12的改编来访问阴道或交配插头的存在或缺失作为交配的指示。</li>
	<li>将母鼠从尾部的底座中抱起, 以短暂地抬起后肢。</li>
	<li>找到交配插头的存在, 一个白色的精物在阴道开口使用牙签。如果交配插头存在, 牙签将无法插入到阴道管。</li>
</ol>3. 组织收集前的准备
<ol><li>准备10毫升的莱博维茨的 L-15 培养基, 辅以1% 胎牛血清 (称为 L15 培养基 + 1% FBS) 在15毫升管。在37° c 的培养基上孵育15分钟的水。</li>
	<li>为了保持组织和精子在下游应用的可行性, 分2毫升的 5ml L15 培养基 + 1% FBS 在2毫升离心管组织收集。</li>
	<li>温暖的阶段显微镜到37° c。将热块设置为37° c, 并在块中放置一个空的 1.5 mL 离心管。</li>
	<li>如果下游应用需要成像的精子运动, 温暖的玻璃幻灯片到37° c。</li>
</ol>4. 组织收集
<ol><li>带 5ml L15 媒体 + 1% FBS 分到组织收集区。</li>
	<li>安乐女性 (~ 08:30h) 使用二氧化碳窒息, 其次是颈椎脱位。 注意: 这是大约 8 h 在交配之后, 假设交配发生在午夜。</li>
	<li>将动物置于仰卧位以暴露腹部区域。</li>
	<li>在后肢附近的腹部喷洒70% 酒精。</li>
	<li>使用解剖剪刀在下腹部的皮肤上做一个小切口 (~ 1 厘米)。</li>
	<li>使用占优势的手拉尾巴 (和后肢, 如果有必要) 而非手拉皮肤相反的方向 (向头部) 暴露腹部肌肉。这种方法是用来减少头发污染的内脏器官的数量。</li>
	<li>将腹部底部的 V 型肌肉切开, 靠近阴道, 露出内脏器官。</li>
	<li>将肠道和腹部脂肪移到侧面以暴露生殖道。</li>
	<li>在阴道区域上方修剪膀胱和其他组织。</li>
	<li>切开耻骨联合以获得阴道组织。</li>
	<li>在阴道开口处提起阴道组织, 用剪刀将阴道导管从直肠组织中分离出来。</li>
	<li>用弹簧剪刀仔细修剪子宫两侧的肠系膜脂肪。 注意: 子宫很脆弱, 里面有精子。注意不要刺穿子宫, 否则精子会丢失。</li>
	<li>抬起较低的生殖道, 并切断两侧的卵巢脂肪垫。</li>
	<li>将整个生殖道转移到5ml 莱博维茨的 L-15 培养基 + 1% FBS。</li>
</ol>5. 从子宫含量采集中测定精子液化 (黏度)
<ol><li>在显微镜的加热阶段, 将组织转化为35毫米无菌组织培养皿, 填充3.5 毫升 5ml L-15 培养基 + 1% FBS, 以清洁血液和其他污染物的组织。 注意: 清洗时, 用脂肪, 而不是子宫来抓组织。此时, 精子有望在子宫内完好无损。</li>
	<li>使用实验室抹布涂抹过量介质的组织。</li>
	<li>在阴道末端放置组织, 同时将卵巢末端置于预热离心管中。在保持阴道末端的同时, 切下子宫底部的子宫角, 允许子宫角落入离心管。</li>
	<li>让精子从子宫角流出管。用无牙钳将子宫角从卵巢末端轻轻挤压到阴道末端, 排出残余的精子。</li>
	<li>丢弃生物危害袋中的组织并焚烧。</li>
	<li>简单地在 minicentrifuge (1-2 秒) 内将精子向下旋转。</li>
	<li>放置管, 平行于加热阶段-精子不会溢出。</li>
	<li>准备好计时器并设置为 "计数"。</li>
	<li>在一个180°角 (平行于管壁) 上插入一个25µL 毛细管进入精子样品。这是为了尽量减少各调查人员之间的差异, 从不同角度持有毛细血管管。</li>
	<li>当毛细管与精子接触时, 立即按下定时器。</li>
	<li>记录精子到达25µL 线所需的时间。</li>
	<li>当测量完成后, 立即把含有精子的管子放回干热块中进行进一步的功能分析, 如对精子运动的视频成像。</li>
	<li>在10% 漂白剂溶液中浸泡20分钟, 并将毛细管放在锋利的容器中, 对含有精子的毛细管进行消毒。</li>
</ol>6. 精子运动的影像学
<ol><li>打开显微镜和视频成像软件。</li>
	<li>吸管20µL 的剩余的精子的5ml 玻璃幻灯片, 而玻璃幻灯片被置于加热阶段。</li>
	<li>盖上玻璃片。</li>
	<li>将滑动片侧放在显微镜下。</li>
	<li>使用20X 物镜来查找感兴趣的区域。</li>
	<li>更改为视频成像的100X 物镜。</li>
	<li>录制的视频在12帧/秒 (fps) 为三十年代。</li>
	<li>随机记录每个动物至少3不同区域的视频。</li>
	<li>快速进行成像, 并尽量减少光线照射到幻灯片, 以保持精子的活力和活力。 注: 此成像程序应在2-3 分钟内完成。</li>
	<li>使用 ImageJ 软件执行数据分析, 如前所述8。</li>
</ol>

<ol>
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A., Lundwall, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Semenogelin,+the+predominant+protein+in+human+semen.+Primary+structure+and+identification+of+closely+related+proteins+in+the+male+accessory+sex+glands+and+on+the+spermatozoa.">Semenogelin, the predominant protein in human semen. Primary structure and identification of closely related proteins in the male accessory sex glands and on the spermatozoa.</a> J Biol Chem. 264 (3), 1894-1900 (1989).</li><li>Veveris-Lowe, T. L., Kruger, S. J., Walsh, T., Gardiner, R. A., Clements, J. 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作者没有什么可透露的。

从小鼠子宫收集精子可以提供优于精子分析的优势体外, 因为前者可以说明精子与女性生殖道分泌物之间的生理相互作用.在这项研究中提出的技术使研究人员能够确定精子的质量, 以及在理想的生理条件下的活力和活力。此外, 还有各种各样的下游分析, 可以使用从子宫收集的精子, 例如, 精子可以计数, 以确定实际精子数进入子宫和精子也可以成像的运动化验。理想的时间点收集精子?...

雌性生殖道由上部和下层组成。上生殖道包括输卵管、子宫和宫颈。下生殖道包括 ectocervix 和阴道。在人类, 被射出的精子沉积在阴道前壁, 毗邻 ectocervix1。然而, 在小鼠体内, 精子在交配2后几分钟内就会被扫入子宫。
精子含有精凝胶, 在射精的几秒钟内凝固, 导致精子被诱捕并固定在3。液化释放的固定精子通过改变精子从凝胶状到水样的一致性。这一过程对精子的运动和哺乳动物的繁殖至关重要。关于精子液化的知识主要是基于体外研究, 其中精子在培养皿中液化。在人类中, 前列腺特异抗原或 PSA (也称为激肽释放酶相关的肽3或 KLK3) 的酵素活性是通过水解 semenogelins (在精子中存在凝胶形成的蛋白质) 来液化的主要贡献者4 ,5,6。降解 semenogelins 释放精子从精物, 增加精子的流动性。自由精子游向输卵管, 使卵子受精。液化 (或精子粘度) 试验是标准的初步筛查精子分析的生殖诊所, 以评估男性生育能力7。
最近发表的一篇文章表明 , 从雌性小鼠生殖道 post - mating 中收集到的的分析可以用来说明液化的不足 , 从而导致生育缺陷8。有缺陷的液化, 如精子高, 导致 11.8-32.3% 的男性不育症患者的9, 表明正常的液化是必不可少的哺乳动物繁殖。然而, 对液化缺陷的研究和处理完全集中在男性7。女性生殖道在精子液化中发挥作用的可能性尚未得到探讨。因此, 收集和测定液化时间的方法将为研究人员提供一种新的诊断工具, 以确定液化是否可能是模型机体中不育的原因之一。

<table><tbody><tr><td>Toothpick</td><td>Diamond</td><td>750-Flat</td><td>Autoclaved sterile, use blunted toothpick. Do not use point-ended</td></tr><tr><td>Dissecting scissors</td><td>Fisher</td><td>08-940</td><td></td></tr><tr><td>Spring scissors</td><td>Roboz</td><td>RS-5600</td><td></td></tr><tr><td>Fine forceps</td><td>Roboz</td><td>RS-5045</td><td></td></tr><tr><td>25-&amp;mu;L glass capillary tube</td><td>VWR</td><td>53432-761</td><td></td></tr><tr><td>Leibovitz&amp;rsquo;s L-15 media</td><td>Life Technologies</td><td>11415064</td><td>Supplement with 1% FBS</td></tr><tr><td>Fetal bovine serum (FBS)</td><td>Gemini Bio-Products</td><td>100-106</td><td></td></tr><tr><td>1.5-mL microcentrifuge tube</td><td>Axygen</td><td>MCT-150-C</td><td></td></tr><tr><td>2-mL microcentrifuge tube</td><td>Axygen</td><td>MCT-200-A</td><td></td></tr><tr><td>Digital dry block heater</td><td>VWR</td><td>13259-052</td><td>For warming the microcentrifuge tube prior to semen collection</td></tr><tr><td>Minicentrifuge</td><td>Corning</td><td>LSE 6765</td><td></td></tr><tr><td>35-mm culture dish</td><td>VWR</td><td>10861-656</td><td></td></tr><tr><td>15-mL polypropylene centrifuge tube</td><td>VWR</td><td>10025-686</td><td></td></tr><tr><td>Waterbath</td><td>Precision</td><td>TSGP05</td><td></td></tr><tr><td>Heated stage stereomicroscope</td><td>Leica Microsystems</td><td>MZ10F</td><td>To keep the tissue warm prior to semen collection</td></tr><tr><td>Brightfield microscope</td><td>Leica Microsystems</td><td>DMi8</td><td>Optional. For video imaging of the sperm motility</td></tr><tr><td>Camera</td><td>Leica Microsystems</td><td>DMC2900</td><td>Optional. For video imaging of the sperm motility</td></tr><tr><td>Glass slides</td><td>Fisher</td><td>12-552-3</td><td>Optional. For video imaging of the sperm motility</td></tr><tr><td>Coverslip</td><td>VWR</td><td>48393-059</td><td>Optional. For video imaging of the sperm motility</td></tr><tr><td>Bleach</td><td></td><td></td><td>Dilute in dH&lt;sub&gt;2&lt;/sub&gt;O to 10% concentration</td></tr></tbody></table>

collection of post-mating semen from the female reproductive tract and measurement of semen liquefaction in mice

在小鼠体内, 被射出的精子在子宫内沉积。射精后, 精子的一致性从凝胶状变为水样, 这一过程称为液化。在这项研究中, 我们展示了如何收集 post-ejaculated 精子从女性生殖道在一个小鼠模型。首先, 发情阶段的成年雌性老鼠在一夜之间被安置在一个雄性笼子里。第二天早上, 交配被证实的交配插头在阴道开口。交配插头的雌性小鼠被安乐死, 每个生殖道都被收集成一个整体 (阴道、子宫、输卵管、卵巢), 确保一个封闭的系统来控制精子。生殖道被放置在1.5 毫升离心管, 阴道被切断, 以释放的精子进入管。为了保证最大的精子量进行分析, 用无牙钳将子宫角从卵巢端挤压到阴道末端, 排出残余的精子。整个生殖道被丢弃。该精子管被短暂剥离。25μ l 毛细管吸管被放置在一个180°角 (平行于管壁) 管。记录了在25μ l 线上填充毛细管所用的时间。从一个成熟的雄性饲养者的精子通常需要大约60-180 秒来填满25μ l 毛细管。这种精子收集技术也可用于其他下游应用, 如精象和运动分析。

在与肥沃的 ctrl 雄性交配后, 从 ctrl 和高雌鱼身上收集了大约8小时的精子。液化 (或精子粘度) 是量化的时间采取填补25μ l 毛细管与精子。从 CTRL 子宫收集的精子采取了2.06 ±0.12 分钟 (mean±东南亚 M, n = 5) 填充毛细管 (图 1)。然而, 从高子宫采集的精子的试验时间超过60分钟 (60.0 ±0.0, mean±东南亚米, n = 5)。这些数据表明, 与高子宫相比, 在 CTRL 的时候, 精子的液化程...

Watch this Scientific Journal Video about Collection of Post-mating Semen from the Female Reproductive Tract and Measurement of Semen Liquefaction in Mice at JoVE.com

Collection of Post-mating Semen from the Female Reproductive Tract and Measurement of Semen Liquefaction in Mice

雌性生殖道交配后精子的收集及对小鼠精子液化的测定

精子的液化需要从精凝胶中解放出来。本研究提供了从雌性生殖道 post-coitus 中收集精子, 以及测定精子液化时间的程序。

In mice, ejaculated semen is deposited in the uterus. After ejaculation, the semen changes consistency from gel-like to watery, a process called ...

collection-post-mating-semen-from-female-reproductive-tract

Research

JoVE Journal

Developmental Biology

16.4K 次观看.  Washington State University. 精子的液化需要从精凝胶中解放出来。本研究提供了从雌性生殖道 post-coitus 中收集精子, 以及测定精子液化时间的程序。

视频: Collection of Post-mating Semen from the Female Reproductive Tract and Measurement of Semen Liquefaction in Mice

Protocols for Vaginal Inoculation and Sample Collection in the Experimental Mouse Model of Candida vaginitis

Vulvovaginal candidiasis (VVC), caused by Candida species, is a fungal infection of the lower female genital tract that affects approximately 75% of otherwise healthy women during their reproductive years18,32-34. Predisposing factors include antibiotic usage, uncontrolled diabetes and disturbance in reproductive hormone levels due to pregnancy, oral contraceptives or hormone replacement therapies33,34. Recurrent VVC (RVVC), defined as three or more episodes per year, affects a separate 5 to 8% of women with no predisposing factors33.
An experimental mouse model of VVC has been established and used to study the pathogenesis and mucosal host response to Candida3,4,11,16,17,19,21,25,37. This model has also been employed to test potential antifungal therapies in vivo13,24. The model requires that the animals be maintained in a state of pseudoestrus for optimal Candida colonization/infection6,14,23. Under such conditions, inoculated animals will have detectable vaginal fungal burden for weeks to months. Past studies show an extremely high parallel between the animal model and human infection relative to immunological and physiological properties3,16,21. Differences, however, include a lack of Candida as normal vaginal flora and a neutral vaginal pH in the mice.
Here, we demonstrate a series of key methods in the mouse vaginitis model that include vaginal inoculation, rapid collection of vaginal specimens, assessment of vaginal fungal burden, and tissue preparations for cellular extraction/isolation. This is followed by representative results for constituents of vaginal lavage fluid, fungal burden, and draining lymph node leukocyte yields. With the use of anesthetics, lavage samples can be collected at multiple time points on the same mice for longitudinal evaluation of infection/colonization. Furthermore, this model requires no immunosuppressive agents to initiate infection, allowing immunological studies under defined host conditions. Finally, the model and each technique introduced here could potentially give rise to use of the methodologies to examine other infectious diseases of the lower female genital tract (bacterial, parasitic, viral) and respective local or systemic host defenses.

Protocols for Vaginal Inoculation and Sample Collection in the Experimental Mouse Model of Candida vaginitis

Key techniques to be used in the evaluation of Candida vaginitis in an experimental animal model are described. The methods will allow rapid collection of vaginal specimens and lymphocytes from draining lumbar lymph nodes. These techniques could give rise to mouse models of other diseases in the female lower genital tract.

实验小鼠模型阴道接种和样品采集的协议念珠菌阴道炎

Vulvovaginal candidiasis (VVC), caused by Candida species, is a fungal infection of the lower female genital tract that affects approximately 75% of ...

科研

免疫与感染

Assessment of Sexual Behavior of Male Mice

Sexual behavior is highly species-specific. Although rodents have slightly different sexual behaviors, mice and rats have a similar sexual behavioral pattern. The purpose of this article is to describe the hormone-induced estrus ovariectomized female model and the experimental procedure for the assessment of sexual behavior of male mice. The most important sexual behavioral elements are demonstrated in the video and illustrations. The critical steps, advantages, and limitations of the sexual behavior test are explained as well. Finally, the behavior parameters are presented, and mounting, intromission, and ejaculation processes in mating are distinguished. Behavioral parameters are assessed in terms of the occurred duration and counts during the test period.

This article describes how to perform sexual behavior tests in male mice.

雄性小鼠性行为评估

Sexual behavior is highly species-specific. Although rodents have slightly different sexual behaviors, mice and rats have a similar sexual behavioral ...

行为学

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 ...

Biaxial Basal Tone and Passive Testing of the Murine Reproductive System Using a Pressure Myograph

The female reproductive organs, specifically the vagina and cervix, are composed of various cellular components and a unique extracellular matrix (ECM). Smooth muscle cells exhibit a contractile function within the vaginal and cervical walls. Depending on the biochemical environment and the mechanical distension of the organ walls, the smooth muscle cells alter the contractile conditions. The contribution of the smooth muscle cells under baseline physiological conditions is classified as a basal tone. More specifically, a basal tone is the baseline partial constriction of smooth muscle cells in the absence of hormonal and neural stimulation. Furthermore, the ECM provides structural support for the organ walls and functions as a reservoir for biochemical cues. These biochemical cues are vital to various organ functions, such as inciting growth and maintaining homeostasis. The ECM of each organ is composed primarily of collagen fibers (mostly collagen types I, III, and V), elastic fibers, and glycosaminoglycans/proteoglycans. The composition and organization of the ECM dictate the mechanical properties of each organ. A change in ECM composition may lead to the development of reproductive pathologies, such as pelvic organ prolapse or premature cervical remodeling. Furthermore, changes in ECM microstructure and stiffness may alter smooth muscle cell activity and phenotype, thus resulting in the loss of the contractile force.
In this work, the reported protocols are used to assess the basal tone and passive mechanical properties of the nonpregnant murine vagina and cervix at 4-6 months of age in estrus. The organs were mounted in a commercially available pressure myograph and both pressure-diameter and force-length tests were performed. Sample data and data analysis techniques for the mechanical characterization of the reproductive organs are included. Such information may be useful for constructing mathematical models and rationally designing therapeutic interventions for women&#8217;s health pathologies.

This protocol utilized a commercially available pressure myograph system to perform pressure myograph testing on the murine vagina and cervix. Utilizing media with and without calcium, the contributions of the smooth muscle cells (SMC) basal tone and passive extracellular matrix (ECM) were isolated for the organs under estimated physiological conditions.

使用压力表谱对毛质生殖系统的双轴基底音和被动测试

The female reproductive organs, specifically the vagina and cervix, are composed of various cellular components and a unique extracellular matrix (ECM). ...

生物工程

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

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-&beta;-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.

Here, we describe how to identify the stage of the murine reproductive (proestrus, estrus, metestrus, or diestrus) by simple, non-invasive collection and cytological assessment of vaginal smear samples. We further describe how vaginal cytology reflects circulating hormonal levels underlying transition through the murine reproductive cycle.

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 ...

生物学

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 ...

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. ...

<meta charset="utf8"></head><body>新型子宫内膜异位症模拟小鼠模型的开发

Establishment of an Experimental Mouse Model of Endometrioma to Study its Related Infertility

Endometrioma (OMA), a subtype of endometriosis characterized by the formation of endometriotic cysts in the ovaries, affects 17-44% of individuals diagnosed with endometriosis. Women with OMA often experience compromised fertility, yet the exact mechanisms underlying OMA-associated infertility remain unclear. Notably, existing animal models simulate superficial peritoneal endometriosis (SUP) and deep infiltrating endometriosis (DIE), leaving a notable gap in research focused on OMA. In response to the gap of knowledge, this paper introduces a pioneering OMA-simulating mouse model and provides a comprehensive description of the techniques and procedures employed in the model. With a high success rate of 83% and ovarian lesion specificity, this model holds significant promise for advancing our understanding of OMA, particularly in the context of infertility. It offers a valuable platform for conducting targeted research into OMA-associated fertility challenges, potentially paving the way for improved diagnostic and therapeutic strategies in the field of reproductive medicine.

<meta charset="utf8"></head><body>建立子宫内膜异位症实验小鼠模型以研究其相关的不孕症

This study introduces a novel mouse model specifically designed to simulate endometrioma, a clinically significant subtype of endometriosis. By utilizing C57BL/6J mice, the model aims to elucidate the pathophysiological mechanisms underlying endometrioma-related infertility, offering a refined tool to bridge the current knowledge gap in reproductive medicine.

雌性生殖道交配后精子的收集及对小鼠精子液化的测定

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此视频中的章节

实验小鼠模型阴道接种和样品采集的协议念珠菌阴道炎

雄性小鼠性行为评估

B组的小鼠模型

使用压力表谱对毛质生殖系统的双轴基底音和被动测试

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

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

建立子宫内膜异位症实验小鼠模型以研究其相关的不孕症

建立子宫内膜异位症实验小鼠模型以研究其相关的不孕症

建立子宫内膜异位症实验小鼠模型以研究其相关的不孕症