鼠前列腺微解剖和手术去势
Summary
该原稿描述了在实验室小鼠前列腺显微解剖和手术去势的协议。我们还通过描绘这些协议产生代表性的结果。最后，我们讨论的优点和这些协议的利用率。
Abstract
Mouse models are used extensively to study prostate cancer and other diseases. The mouse is an excellent model with which to study the prostate and has been used as a surrogate for discoveries in human prostate development and disease. Prostate micro-dissection allows consistent study of lobe-specific prostate anatomy, histology, and cellular characteristics in the absence of contamination of other tissues. Testosterone affects prostate development and disease. Androgen deprivation therapy is a common treatment for prostate cancer patients, but many prostate tumors become castration-resistant. Surgical castration of mouse models allows for the study of castration resistance and other facets of hormonal biology on the prostate. This procedure can be coupled with testosterone reintroduction, or hormonal regeneration of the prostate, a powerful method to study stem cell lineages in the prostate. Together, prostate micro-dissection and surgical castration opens up a multitude of opportunities for robust and consistent research of prostate development and disease. This manuscript describes the protocols for prostate micro-dissection and surgical castration in the laboratory mouse.
Introduction
前列腺是癌症在美国男性中最常见的部位。近22人，每年将被诊断患有前列腺癌，并且约27,000人会屈服于他们的疾病1。男人有美国1前列腺癌诊断的1/7的寿命风险。良性前列腺增生（BPH），前列腺的年龄有关的非癌性增大，也是一个普遍的状况，影响男性超过80％，超过80 2，因此，前列腺是相当多的研究的重点。
小鼠模型已被广泛用来研究前列腺3的疾病。总体而言，小鼠前列腺是人类腺4的优秀代表，但也有相似性和小鼠和人前列腺解剖学和生理学5之间的差异。在这两个物种中，前列腺位于膀胱的基础上，环绕尿道。人类前列腺是一个LO定，分为四个区域：中部，过渡，外设和前纤维肌性基质。与此相反，在小鼠前列腺包括在尿道周围的不同位置三次配对裂片：前壁，腹和背外侧。在细胞水平上，主要的区别在于，基底细胞至管腔细胞比在人为约1：1，但它是1：4小鼠6。鼠基质也是相对于人类的小鼠不同 – 人类有更广泛的平滑肌，而肌肉层是在小鼠前列腺薄得多。正确的识别，解剖和小鼠前列腺的处理是小鼠前列腺的研究是必不可少的。
激素水平显着影响在人类和小鼠中的前列腺的发育和内环境稳定。而雌激素似乎在前列腺发育7发挥作用，在前列腺的最重要的激素是睾酮。睾酮是腺发展的重要d维持。以下物理或化学去势，在成人前列腺apoptose的管腔细胞的约90％，和腺收缩。如果睾丸激素去势条件下重新引入个人，前列腺能自己再生满负荷生产。睾酮也似乎驱动前列腺癌，这是为什么雄激素剥夺疗法是一种常用的治疗策略。然而，许多前列腺癌成为去势抵抗性。此外，男性接受雄激素剥夺治疗前列腺癌的治疗经历的阉割和再生条件的周期。在小鼠中，手术去势，通过重新引入睾酮前列腺激素再生沿，是一个重要的工具，使用它来研究去势抗性和睾酮循环对前列腺的作用。
在本文中，我们将讨论并演示通过其中LOCAT适当的技术e和微解剖小鼠前列腺，以及通过外科手术阉割鼠标。
Protocol
Representative Results
Discussion
前列腺显微解剖允许特定瓣实验和小鼠前列腺的分析（ 图1）。在遗传工程小鼠模型，表型可能在未在其他见过叶中看到。此外，对于组织学分析，该协议确保了纯的前列腺组织的最大量可以被切片，染色，而不存在于所述部分中的其他泌尿生殖道的组织。最后，对于单细胞实验，该协议允许几乎纯质的前列腺细胞的分离。上皮细胞富集试剂盒可以进一步净化从基质细胞前列腺细胞。…
Declarações
The authors have nothing to disclose.
Acknowledgements
一些在本手稿的数字是慷慨巴特O.威廉姆斯的实验室提供。作者由美国国家癌症研究所授予U54CA143803，CA163124，CA093900，和CA143055支持。
Materials
| Surgical tools | Roboz | Various | |
| Formaldehyde | Sigma | F8775 | |
| OCT medium | VWR | 25608-930 | Must be placed on dry ice to freeze, and stored at -80C |
| PBS | Life Technologies | 10010 | |
| Isoflurane | Johns Hopkins | Also available from vendors online | |
| Cautery Pen | Medline | ESCT002 | |
| Silk suture | AD Surgical | M-S330R19 | |
| Surgical Wound Clips | Roboz | RS-9265 |
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