小鼠睾丸切除术和卵巢切除术减少性激素的产生
Summary
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。
Abstract
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。
Introduction
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。
Protocol
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 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。 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。…
Representative Results
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…
Discussion
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、?…
Offenlegungen
The authors have nothing to disclose.
Acknowledgements
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。
Materials
| 1mL Syringe | BD | 309659 | |
| 30G 1/2" Needle | BD | 305106 | |
| AutoClip System | Fine Science Tools | 12020-00 | |
| Betadine Solution | Fisher Scientific | NC0158124 | |
| Cotton-Tipped Applicators | Fisher Scientific | 10-000-692 | |
| Double -ended Micro Spatula | Fine Science Tools | 10091-12 | |
| Galilean Loupes | Fine Science Tools | 28050-30 | Optional, can provide better clarity during procedure |
| Gauze Sponges, 4"x4" | Fisher Scientific | 13-761-52 | |
| Graefe Forceps | Fine Science Tools | 11150-10 | |
| High Temp Cautery Kit | Fine Science Tools | 18010-00 | Using the fine tip attachment |
| Needle Holders | Fine Science Tools | 12001-13 | |
| PGA Absorbable Suture:4-0 / NFS-2 Reverse Cutting 19MM / 30 IN | Covetrus | 29242 | 4-0 or 5-0 Absorbable sutures are best |
| Rodent Warming pad | Kent Scientific | RT-0515 | |
| Sterile Alcohol Prep Pads | Fisher Scientific | 22-363-750 | |
| Straight Locking Micro Needle Holders | Fine Science Tools | 12060-01 | |
| Surgical Scissors | Fine Science Tools | 140-60-09 | |
| Vannas Spring Scissors – 2.5mm Cutting Edge | Fine Science Tools | 15000-08 | |
| Veet Sensitive Hair Remover Gel Cream | Amazon | N/A | |
| Wahl Professional Animal Compact Trimmer and Grooming Kit, Blue | Amazon | #9861-900 |
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Diesen Artikel zitieren
Rowe, A. A., Issioui, Y., Johnny, B., Wert, K. J. Murine Orchiectomy and Ovariectomy to Reduce Sex Hormone Production. J. Vis. Exp. (201), e64379, doi:10.3791/64379 (2023).