Name: murine orchiectomy and ovariectomy to reduce sex hormone production
Uploaded: 2023-11-17T14:40:00
Description: Watch this Scientific Journal Video about Murine Orchiectomy and Ovariectomy to Reduce Sex Hormone Production at JoVE.com

我们感谢德克萨斯大学西南医学中心动物资源中心在外科培训和方案审查方面的帮助。我们感谢 Wert Lab 支持团队提供的宝贵帮助。这项工作得到了美国国立卫生研究院（NIH P30EY030413）的资金支持。Biorender.com 用于创建卡通原理图。

所有动物实验均由UTSW机构动物护理和使用委员会批准（APN#2019-102840）。
1. 鼠睾丸切除术
<ol><li>准备无菌工作区域，并确保必要的手术器械已经过消毒并随时可用。遵循啮齿动物生存手术的无菌实践。</li>
	<li>记录雄性小鼠的体重，并根据机构指南施用首选的麻醉方法。在该方案中，通过精密汽化器输送 2% 异氟醚以维持麻醉。手术前，根据机构指南服用止痛药。...

<ol>
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D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+early+bilateral+ovariectomy+in+mice+as+a+model+of+left+heart+disease.">Evaluation of early bilateral ovariectomy in mice as a model of left heart disease.</a> American Journal of Physiology Heart-Circulartory Physiology. 322 (6), H1080-H1085 (2022).</li><li>Lu, H., Ma, K., Jin, L., Zhu, H., Cao, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=17&#946;-estradiol+rescues+damages+following+traumatic+brain+injury+from+molecule+to+behavior+in+mice.">17&#946;-estradiol rescues damages following traumatic brain injury from molecule to behavior in mice.</a> Journal of Cell Physiology. 233 (2), 1712-1722 (2018).</li><li>Meydan, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+testosterone+on+orchiectomy-induced+oxidative+damage+in+the+rat+hippocampus.">Effects of testosterone on orchiectomy-induced oxidative damage in the rat hippocampus.</a> Journal of Chemical Neuroanatomy. 40 (4), 281-285 (2010).</li><li>Ohlson, N., Bergh, A., Persson, M. 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手术切除睾丸和卵巢允许在受控激素剥夺下研究小鼠生理学。该技术对许多科学领域都很重要，包括神经退行性疾病、矿物质代谢、心血管和生殖健康 15,16,17,18,19,20,21。在这里，我们详细介绍了使用生存手术快速、安全、?...

睾丸和卵巢是负责产生性激素的主要器官。导致睾酮和雌激素产生的荷尔蒙通讯级联是一个特征明确的过程，始于下丘脑，释放促性腺激素释放激素 （GnRH）1。GnRH的释放导致垂体释放黄体生成素（LH）和卵泡刺激素（FSH）。当这些激素进入血液时，它们会影响身体的其他组织。LH 的主要目标是睾丸（男性）和卵巢（女性）2。作为对LH的反应，睾丸产生并释放睾酮3。同样，卵巢产生雌激素4。虽然这些激素的预期效果是使细胞和身体为受精做好准备并确保生殖系统正常运作，但许多其他身体系统也会受到影响。
性激素与多种生理功能有关。例如，雌激素通过防止破骨细胞吸收骨骼来帮助维持骨稳态。因此，卵巢切除小鼠模型可用于研究骨质疏松症等骨骼疾病的生理学5,6,7。睾酮和雌激素也是许多心血管和神经退行性疾病的研究靶点。最近，睾丸激素分泌升高加上高脂肪饮食与血管氧化应激有关8。在大脑中，卵巢切除术后 LH 的变化导致了空间记忆的改变9。卵巢切除术后雌激素的减少也已成为研究海马细胞死亡的模型系统，因为这会诱导细胞凋亡，导致记忆缺陷10。睾酮在肾移植后的小鼠模型和人类肾脏生长中也显示出作用11。
激素剥夺小鼠模型的创建允许研究性激素及其激素级联对各种疾病或组织的影响。这可以通过手术切除睾丸（睾丸切除术）或卵巢（卵巢切除术）来实现。该过程可以在任何品系的小鼠中进行，当它们与断奶年龄（二十一天）或任何成年年龄一样小。卵巢切除术在雌性小鼠中进行，而睾丸切除术在雄性小鼠中进行。通过去除这些器官，雌激素和睾丸激素及其许多衍生物（如黄体酮）的水平可以大大降低12,13。通过适当的技术，在小鼠中进行睾丸切除术或卵巢切除术的过程可以是快速且微创的。以安全有效的方式快速切除这些器官可以实现快速的手术处理，同时在正确执行时具有 100% 的存活率，从而将小鼠数量保持在最低限度。在这里，我们详细介绍了快速切除睾丸和卵巢的方案，并展示了适当的术后监测，使研究人员能够快速安全地进行这种手术。我们还包括性器官和周围组织的视觉示例，以便在执行此手术时为外科医生提供解剖标志。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1mL Syringe</td>
			<td>BD</td>
			<td>309659</td>
			<td></td>
		</tr>
		<tr>
			<td>30G 1/2&#34; Needle</td>
			<td>BD</td>
			<td>305106</td>
			<td></td>
		</tr>
		<tr>
			<td>AutoClip System</td>
			<td>Fine Science Tools</td>
			<td>12020-00</td>
			<td></td>
		</tr>
		<tr>
			<td>Betadine Solution</td>
			<td>Fisher Scientific</td>
			<td>NC0158124</td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton-Tipped Applicators</td>
			<td>Fisher Scientific</td>
			<td>10-000-692</td>
			<td></td>
		</tr>
		<tr>
			<td>Double -ended Micro Spatula</td>
			<td>Fine Science Tools</td>
			<td>10091-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Galilean Loupes</td>
			<td>Fine Science Tools</td>
			<td>28050-30</td>
			<td>Optional, can provide better clarity during procedure</td>
		</tr>
		<tr>
			<td>Gauze Sponges, 4&#34;x4&#34;</td>
			<td>Fisher Scientific</td>
			<td>13-761-52</td>
			<td></td>
		</tr>
		<tr>
			<td>Graefe Forceps</td>
			<td>Fine Science Tools</td>
			<td>11150-10</td>
			<td></td>
		</tr>
		<tr>
			<td>High Temp Cautery Kit</td>
			<td>Fine Science Tools</td>
			<td>18010-00</td>
			<td>Using the fine tip attachment</td>
		</tr>
		<tr>
			<td>Needle Holders</td>
			<td>Fine Science Tools</td>
			<td>12001-13</td>
			<td></td>
		</tr>
		<tr>
			<td>PGA Absorbable Suture:4-0 / NFS-2 Reverse Cutting 19MM / 30 IN</td>
			<td>Covetrus</td>
			<td>29242</td>
			<td>4-0 or 5-0 Absorbable sutures are best</td>
		</tr>
		<tr>
			<td>Rodent Warming pad</td>
			<td>Kent Scientific</td>
			<td>RT-0515</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile Alcohol Prep Pads</td>
			<td>Fisher Scientific</td>
			<td>22-363-750</td>
			<td></td>
		</tr>
		<tr>
			<td>Straight Locking Micro Needle Holders</td>
			<td>Fine Science Tools</td>
			<td>12060-01</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical Scissors</td>
			<td>Fine Science Tools</td>
			<td>140-60-09</td>
			<td></td>
		</tr>
		<tr>
			<td>Vannas Spring Scissors - 2.5mm Cutting Edge</td>
			<td>Fine Science Tools</td>
			<td>15000-08</td>
			<td></td>
		</tr>
		<tr>
			<td>Veet Sensitive Hair Remover Gel Cream</td>
			<td>Amazon</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>Wahl Professional Animal Compact Trimmer and Grooming Kit, Blue&#160;</td>
			<td>Amazon</td>
			<td>#9861-900</td>
			<td></td>
		</tr>
	</tbody>
</table>

murine orchiectomy and ovariectomy to reduce sex hormone production

性激素信号转导在多个器官系统以及各种疾病（包括神经退行性疾病）的进展中起着关键作用。在小鼠模型系统中操纵性激素水平可以研究它们对器官/组织和疾病进展的影响。睾丸切除术 - 睾丸切除术 - 卵巢切除术 - 卵巢切除术 - 提供了一种消耗内源性激素的方法，以便可以通过药物或其他递送方法提供精确的激素水平。在这里，我们为小鼠模型系统中的睾丸切除术和卵巢切除术提供了快速和微创的方法，以减少性激素。该方案详细介绍了手术准备和通过阴囊切除睾丸，以及通过左右背侧的两个切口切除卵巢。

这里介绍的程序是在C57BL / 6J背景中的一至三个月大的小鼠中进行的。雄性小鼠在手术时体重为16-28克，雌性小鼠体重为14-24克。该程序已经过优化，适用于从断奶到成年的许多年龄的小鼠。
手术睾丸切除术涉及腹侧阴囊中的单个皮肤切口，如图 1A 所示。两个睾丸一次切除一个，并通过输精管和精索血管切断，导致睾丸和附睾的切除（图1B...

Watch this Scientific Journal Video about Murine Orchiectomy and Ovariectomy to Reduce Sex Hormone Production at JoVE.com

Murine Orchiectomy and Ovariectomy to Reduce Sex Hormone Production

这份手稿描述了一种快速进行存活啮齿动物睾丸切除术和卵巢切除术的一致方法。

小鼠睾丸切除术和卵巢切除术减少性激素的产生

Sex hormone signaling plays a critical role in multiple organ systems as well as in the progression of various diseases, including neurodegenerative ...

该手术的总体目标是证明以快速、安全和有效的方式手术切除小鼠睾丸和卵巢。小鼠睾丸切除术是通过在阴囊中切腹中线切口进行的，睾丸被隔离并切除，然后闭合皮肤切口。小鼠卵巢切除术是通过在腹部进行背外侧切口，在那里卵巢被隔离和切除，然后闭合皮肤切口，可以选择在另一侧重复此手术以进行双侧卵巢切除术。要开始小鼠睾丸切除术，首先要记录雄性小鼠的体重，并准备按照您的机构指南使用适当的止痛药和麻醉剂。一旦鼠标处于麻醉状态，用润滑眼凝胶覆盖眼睛，以保护它们在手术过程中不会变干。在继续之前，通过执行脚趾捏反应测试，确保鼠标处于适当的麻醉平面。接下来，剃掉鼠标的腹股沟和阴囊区域。使用棉签涂抹器，在手术部位和周围区域涂抹一层薄薄的脱毛膏。30 秒后，用干净的纱布海绵去除脱毛膏。将鼠标转移到手术区域并确保有热源可用。使用柔软的胶带将鼠标的脚粘在手术垫上。如果睾丸已经上升到腹部，对腹部轻轻按压以帮助它们下降。用Betadine溶液擦洗皮肤，从手术部位的中心开始，然后向外轻松向剃须区域的边缘工作。在Betadine磨砂之后，以类似的方式进行酒精磨砂。重复此过程三次以确保适当的皮肤消毒。用手术单盖住小鼠。抓住皮肤，做一个长约 1 至 1 1/2 厘米的中线切口。抓住皮肤的切割边界，使用钝头器械将皮肤与下面的组织分开。花点时间在切口的外侧、上缘和下缘上做这个。接下来，通过将皮肤切口滑动到直接顶部来隔离一个睾丸。用一把镊子抓住睾丸周围薄而透明的提肌。正确组织的识别可以通过其透明度和独立于其内部的底层组织移动的能力来确认。接下来，通过在组织的顶端切开一个切口来切开提睾肌。将锁定夹放在切割的提骨肌的后缘上，以保持组织上的张力。用一把镊子抓住睾丸并将其从其乳鞘中取出。睾丸取出后，将组织横向滚动，并将连接睾丸肌肉与睾丸和附睾的睾丸和附睾的 cremaster 插入点定位在睾丸后侧。夹住并撕裂此组织以切断此连接。慢慢拉出腹股沟脂肪垫，使其外部化。在这样做时，请注意出血的迹象，并确保在此过程中不要伤害任何血管。接下来，使用一对锁定止血器夹住精索和血管。在睾丸保持轻微张力的同时，烧灼止血钳夹远端的精索和血管。缓慢释放止血剂，确保组织残端止血良好，不会继续出血。对 cremaster 肌肉施加轻微的向外压力将使该组织残端缩回体腔。将提睾肌的两侧并拢。使用 4-0 或 5-0 可吸收缝合线，放置缝合线以连接 cremaster 肌肉的两端。缝合完成后，将缝合材料的末端修剪成 1/2 厘米长。通过将皮肤切口滑动到另一侧的顶部，对另一个睾丸重复此过程，并按照用于切除第一个睾丸的相同步骤进行操作。一旦两个睾丸都被切除，并且已经缝合了cremaster的两侧，将皮肤向上剥离，并准备在该区域使用伤口夹。确保缝合线末端不会卡在伤口夹中。鼠标现在可以继续进行术后监测了。为了准备小鼠卵巢切除术，称量雌性小鼠的体重，并按照您的机构指南给予止痛药和麻醉。涂抹润滑眼凝胶并检查以确保鼠标没有脚趾捏合反应。剃掉小鼠背部的背外侧区域以准备手术部位。使用棉签涂抹器，在剃须区域涂抹一层脱毛膏。30 秒后，用干净的纱布海绵去除乳霜。将鼠标转移到手术垫上，并确保有热源可用。用Betadine溶液擦洗皮肤，从中心开始，然后向外向剃须区域的边缘涂抹。然后用类似方式涂抹 70% 酒精溶液擦洗皮肤。重复此过程总共三次，以确保适当的皮肤消毒。按照书面方案中的详细说明计划手术切口。将手术单放在小鼠的手术部位上。用镊子抓住皮肤，用一厘米的切口切开皮肤。使用钝头器械或普通止血剂将皮肤与下层组织分开。花点时间在切口的外侧、上缘和下缘上做这个。抓住腹壁，切开 1/2 厘米的切口。使用一把镊子将卵巢定位在切口内，然后拉动以使其外部化。一旦确定了组织，使用一对止血剂夹住并压碎远端子宫角。使用第二对止血剂或一把镊子夹住第一个夹子的远端。保持底部止血不动，将顶部夹向上拉并远离身体，以对远端子宫角进行挤压和撕裂。慢慢释放止血剂并观察以确保手术部位没有过多出血。将腹壁的两侧放在一起，并使用带有 4-0 可吸收缝合材料的单条缝合线缝合切口。缝合线固定后，将缝合线材料的末端修剪成 1/2 厘米长。将皮肤的两侧重新组合在一起，并准备放置伤口夹。在剥离皮肤的同时，在切口部位放置一个伤口夹。鼠标现在可以继续进行术后监测了。根据您的机构指南记录手术日期和时间以及镇痛药给药。一旦小鼠接受了手术睾丸切除术或卵巢切除术，将它们转移到干净、加热的笼子里。一旦小鼠恢复胸骨卧位，将它们放在干净的笼子里，并提供湿润的食物和水。在恢复期间，经常检查小鼠的手术部位，寻找任何感染或出血的迹象。为防止损坏伤口夹，请在手术后一周内从卵巢切除小鼠的笼子中移除任何啮齿动物冰屋栖息地。14 天后，从鼠标皮肤上取下伤口夹。演示的方案详细说明了如何在小鼠中进行睾丸切除术和卵巢切除术以生成性激素耗尽模型。这包括通过阴囊中的中线切口切除睾丸，然后将睾丸外化并切断血管中的精索，从而切除睾丸、附睾和腹股沟脂肪垫。手术卵巢切除术可以通过背侧切口进入背部并在卵巢、输卵管和子宫角远端外侧进行。随后切断远端子宫角，导致卵巢和输卵管切除。仔细的手术技术和适当的术后护理将使手术部位愈合良好。这些外科手术可以通过在睾丸切除术后一周分析小鼠血清样本中的激素水平来进一步验证。卵巢切除术可以通过观察卵巢切除后的子宫萎缩来确认。该协议能够快速有效地从小鼠身上切除卵巢和睾丸，以产生性激素耗尽模型。这可以有很多用途来研究这些激素的消耗如何影响各种器官和生理过程。

Research

JoVE Journal

Medicine

The Measurement and Treatment of Suppression in Amblyopia

测量和制止在弱视治疗

Amblyopia, a developmental disorder of the visual cortex, is one of the leading causes of visual dysfunction in the working age population. Current ...

科研

医学

Isolating and Using Sections of Bovine Mesenteric Artery and Vein as a Bioassay to Test for Vasoactivity in the Small Intestine

隔离和使用的牛肠系膜动脉和静脉部分，作为生物测定在小肠测试的Vasoactivity

Mammalian gastrointestinal systems are constantly exposed to compounds (desirable and undesirable) that can have an effect on blood flow to and from that ...

An Ex vivo Model to Study Hormone Action in the Human Breast

An Ex vivo Model to Study Hormone Action in the Human Breast

一个<em&gt;体外</em&gt;模型在乳腺癌研究激素的作用

The study of hormone action in the human breast has been hampered by lack of adequate model systems. Upon in vitro culture, primary mammary epithelial ...

Povidone Iodine Rectal Preparation at Time of Prostate Needle Biopsy is a Simple and Reproducible Means to Reduce Risk of Procedural Infection

聚维酮碘直肠准备在前列腺穿刺活检的时间是一个简单的和可重复的手段，以减少程序感染的风险

Single institution and population-based studies highlight that infectious complications following transrectal ultrasound guided prostate needle biopsy ...

Transplantation of Adipose Tissue-Derived Stem Cell Sheet to Reduce Leakage After Partial Colectomy in A Rat Model

脂肪组织干细胞移植对大鼠部分结肠切除术后渗漏的减少

Anastomotic leakage is a disastrous complication after colorectal surgery. Although current methods for leakage prevention have different levels of ...

A Clinical Trial Assessing the Safety, Efficacy, and Delivery of Olive-Oil-Based Three-Chamber Bags for Parenteral Nutrition

一项临床试验，评估基于橄榄油的三腔袋的亲子营养的安全性、有效性和交付

Limited evidence exists to precisely estimate efficacy and safety differences between parenteral nutrition (PN) prepared using olive-oil-based ...

Mechanisms Underlying Gut Hormone Secretion Using the Isolated Perfused Rat Small Intestine

利用隔离的完美大鼠小肠进行肠道激素分泌的机制

The gut is the largest endocrine organ of the body, producing more than 15 different peptide hormones that regulate appetite and food intake, digestion, ...

Cooling or Warming the Esophagus to Reduce Esophageal Injury During Left Atrial Ablation in the Treatment of Atrial Fibrillation

在心房颤动治疗中，为食道冷却或加热，以减少左心房消融期间的食管损伤

Ablation of the left atrium using either radiofrequency (RF) or cryothermal energy is an effective treatment for atrial fibrillation (AF) and is the most ...

Network Pharmacology Prediction and Experimental Validation of Trichosanthes-Fritillaria thunbergii Action Mechanism Against Lung Adenocarcinoma

Network Pharmacology Prediction and Experimental Validation of Trichosanthes-Fritillaria thunbergii Action Mechanism Against Lung Adenocarcinoma

毛滴虫对肺腺癌作用机制的网络药理学预测及实验验证

We aimed to study the mechanism of Trichosanthes-Fritillaria thunbergii in treating lung adenocarcinoma (LUAD) based on network pharmacology and ...

Production of Modified Autologous Conditioned Serum and Ex Vivo Assessment of Its Healing Potential in Murine Corneal Epithelium

Production of Modified Autologous Conditioned Serum and Ex Vivo Assessment of Its Healing Potential in Murine Corneal Epithelium

改良自体条件血清的生产及其在小鼠角膜上皮中愈合潜力的离 体 评估

Human blood-derived topical therapies have been a boon to clinicians in recent decades. Autologous serum (AS) and platelet-rich plasma (PRP) are enriched ...