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

We thank the University of Texas Southwestern Medical Center Animal Resource Center for their help in surgical training and protocol review. We thank the Wert Lab support team for their invaluable assistance. This work has been supported by funds from the National Institute of Health (NIH P30EY030413). Biorender.com was used for the creation of cartoon schematics.

All animal experiments were approved by the Institutional Animal Care and Use Committee at UTSW (APN#2019-102840).
1. Murine orchiectomy
<ol>
	<li>Prepare an aseptic working field and ensure that necessary surgical instruments have been sterilized and are readily available. Follow aseptic practices for rodent survival surgery.</li>
	<li>Record the weight of the male mouse and administer the preferred method of anesthesia according to the institutional guidelines. In this pro...

<ol>
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Surgical removal of testes and ovaries allows for studying murine physiology under controlled hormone deprivation. This technique is important for many fields of science, including neurodegeneration, mineral metabolism, cardiovascular, and reproductive health15,16,17,18,19,20,21. Here, we de...

The testes and ovaries are the primary organs responsible for&#160;sex hormone production. The cascade of hormonal communication leading to the production of testosterone and estrogen is a well-characterized process that begins in the hypothalamus with the release of gonadotropin-releasing hormone (GnRH)1. The release of GnRH causes the release of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) from the pituitary gland. As these hormones enter the bloodstream, they then affect other tissues in the body. The primary target of LH is the testes (in males) and the ovaries (in females)2. In response to LH, the testes produce and release testosterone3. Similarly, the ovaries produce estrogen4. While the intended effects of these hormones are to prepare the cells and body for fertilization and ensure a functioning reproductive system, many other bodily systems can be affected.
Sex hormones have been linked to several physiological functions. For instance, estrogen helps maintain bone homeostasis by preventing the resorption of bone by osteoclasts. For this reason, ovariectomized mouse models can be used to study the physiology of bone diseases such as osteoporosis5,6,7. Testosterone and estrogen are also research targets for many cardiovascular and neurodegenerative diseases. Recently, elevated testosterone production coupled with high-fat diet has been linked to vascular oxidative stress8. In the brain, changes in LH after ovariectomy have caused alterations in spatial memory9. Reduction of estrogens following ovariectomy has also become a model system for studying cell death in the hippocampus, as this can induce apoptosis, resulting in memory deficits10. Testosterone has also shown a role in the growth of kidneys both in mouse models and humans following kidney transplantation11.
The creation of a hormone-deprived murine model allows for the study of sex hormones and their hormone cascades on various diseases or tissues. This can be accomplished by surgical removal of the testes (orchiectomy) or ovaries (ovariectomy). This procedure can be performed in mice of any strain when they are as young as weaning age (twenty-one days) or any adult age. Ovariectomy is performed in female mice, while orchiectomy is performed in male mice. By removing these organs, the levels of estrogen and testosterone, and many of their derivatives, such as progesterone, can be greatly reduced12,13. The process of performing orchiectomies or ovariectomies in mice can be rapid and minimally invasive with the proper technique. Rapid excision of these organs in a safe and efficient manner can allow for quick surgical processing while keeping mouse numbers minimal by having a 100% survival rate when performed correctly. Here, we detail a protocol for the rapid excision of the testes and ovaries and demonstrate the proper post-surgical monitoring to enable researchers to perform this surgery quickly and safely. We also include visual examples of the sex organs and surrounding tissues to provide the surgeon with anatomical landmarks when performing this procedure.

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murine orchiectomy and ovariectomy to reduce sex hormone production

Sex hormone signaling plays a critical role in multiple organ systems as well as in the progression of various diseases, including neurodegenerative disease. The manipulation of sex hormone levels in the murine model system allows for the study of their impact on organs/tissues and within disease progression. Orchiectomy - the surgical removal of the testes - and ovariectomy - the surgical removal of the ovaries - provide a method to deplete the endogenous sex hormones so that the precise hormone levels can be provided through drug or other delivery methods. Here, we provide rapid and minimally invasive methods for both orchiectomy and ovariectomy in the murine model system for the reduction of sex hormones. This protocol details the surgical preparation and excision of the testes through the scrotal sac, and excision of the ovaries via two incisions in the right and left lateral dorsum.

The procedure presented here is performed in one- to three-month-old mice in the C57BL/6J background. Male mice weighed 16-28 g, and female mice weighed 14-24 g at the time of the procedure. This procedure has been optimized to be applicable for mice of many ages, from weaning through adulthood.
Surgical orchiectomy involves a single skin incision in the ventral scrotal sac, as depicted in Figure 1A. Both testes are removed one at a time and are severed through th...

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

This manuscript describes a consistent way to quickly perform survival rodent orchiectomies and ovariectomies.

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

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