eoe sub articles

EoE Sub Articles

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Create PCOS-like Mouse Model
NOTE: The mice used in this study were a mixed background (C57/B6, CD1, 129Sv) and were maintained with food and water&#160;ad libitum&#160;in a 14/10 h light/dark cycle at 24 &#176;C.
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	<li>DHT Pellet Preparation
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		<li>A...

subcutaneous dihydrotestosterone (dht) treatment: mouse model of polycystic ovary syndrome (pcos)

Source: Xue, P., et al. <a href="https://www.jove.com/video/58379/a-hyperandrogenic-mouse-model-to-study-polycystic-ovary-syndrome">A Hyperandrogenic Mouse Model to Study Polycystic Ovary Syndrome</a>.&#160;J. Vis. Exp. (2018).
This article describes a way to prepare and insert dihydrotestosterone (DHT) pellets to model Polycystic Ovary Syndrome (PCOS) in mice. The example protocol demonstrates the procedure in mice used for reproductive and metabolic studies. &#160;&#160;

Subcutaneous Dihydrotestosterone (DHT) Treatment: Mouse Model of Polycystic Ovary Syndrome (PCOS)

Source: Xue, P., et al. A Hyperandrogenic Mouse Model to Study Polycystic Ovary Syndrome.&#160;J. Vis. Exp. (2018).
This article describes a way to ...

First, to prepare the pellet, inject medical adhesive silicone down one end of a pre-cut piece of silicone tubing. Next, pack dihydrotestosterone or DHT crystal powder tightly inside the tubing. Then, use medical adhesive silicone to seal the other end of the tubing. Pellets should have equal amounts of silicone at each end with no bubbles. Place pellets in saline solution before inserting them in a female mouse.
On an anesthetized mouse, use a cotton swab with disinfectant to clean the shaved surgical area. Make a small incision through the skin near the neck. Use a trochar to make a tunnel under the skin in the direction of the mouse&#39;s nose. Insert the pellet into the tunnel. Lastly, seal the incision with surgical adhesive. The pellet will release a steady dose of DHT and is replaced every 4 weeks to ensure prolonged exposure to the drug.

subcutaneous-dihydrotestosterone-dht-treatment-mouse-model-polycystic

Research

JoVE Encyclopedia of Experiments

Biology

Rodent Models

Adult

Murine Experimental Model of Original Tumor Development and Peritoneal Metastasis via Orthotopic Inoculation with Ovarian Carcinoma Cells

Epithelial ovarian carcinoma (EOC) is associated with a poor prognosis because it shows peritoneal dissemination. To improve the prognosis, it is important to control peritoneal dissemination. However, it is still unclear how tumor cells detach from primary lesions and attach to the mesothelium. The establishment of an appropriate animal model is needed to gain an understanding of the mechanism of peritoneal dissemination in vivo. In the current study, we introduce the process from the local injection of EOC cells into the murine ovarian surface to the development of metastasis, including the peritoneum and distant organs. Female nude mice (BALB/c nu/nu) at 8 weeks of age were used. Under a microscopic field of view, EOC cells (1 x 105 cells/&#181;l of medium-extracellular matrix (ECM)-based hydrogel/unilateral ovary/mouse) were injected into murine ovaries through a retroperitoneal approach from the dorsal flank. This proposed method is a less invasive procedure for the mouse and minimizes damage to the ovary. Here, we describe the methodological steps in the development of the original and metastatic tumor formation of EOC.

Murine Experimental Model of Original Tumor Development and Peritoneal Metastasis via Orthotopic Inoculation with Ovarian Carcinoma Cells

To clarify the multistep process of peritoneal dissemination of ovarian carcinoma, the current study presents a murine experimental model of original tumor development and peritoneal metastasis via orthotopic inoculation with these tumor cells.

Epithelial ovarian carcinoma (EOC) is associated with a poor prognosis because it shows peritoneal dissemination. To improve the prognosis, it is ...

JoVE Journal

Cancer Research

Culture and Co-Culture of Mouse Ovaries and Ovarian Follicles

The mammalian ovary is composed of ovarian follicles, each follicle consisting of a single oocyte surrounded by somatic granulosa cells, enclosed together within a basement membrane. A finite pool of follicles is laid down during embryonic development, when oocytes in meiotic arrest form a close association with flattened granulosa cells, forming primordial follicles. By or shortly after birth, mammalian ovaries contain their lifetime&#8217;s supply of primordial follicles, from which point onwards there is a steady release of follicles into the growing follicular pool.
The ovary is particularly amenable to development in vitro, with follicles growing in a highly physiological manner in culture. This work describes the culture of whole neonatal ovaries containing primordial follicles, and the culture of individual ovarian follicles, a method which can support the development of follicles from an immature through to the preovulatory stage, after which their oocytes are able to undergo fertilization in vitro. The work outlined here uses culture systems to determine how the ovary is affected by exposure to external compounds. We also describe a co-culture system, which allows investigation of the interactions that occur between growing follicles and the non-growing pool of primordial follicles.

This protocol describes the primary culture/co-culture of mouse ovarian tissue, using ovaries from neonatal mice and individual ovarian follicles from prepubertal mice. The culture techniques support development in a highly physiological manner, allowing investigation of the effect of extrinsic agents on the ovary, and of interactions between ovarian follicles.

The mammalian ovary is composed of ovarian follicles, each follicle consisting of a single oocyte surrounded by somatic granulosa cells, enclosed together ...

Developmental Biology

Evaluation of Hepatic Glucose Production in a Polycystic Ovary Syndrome Mouse Model

Polycystic ovary syndrome (PCOS) is a common disease that results in disorders of glucose metabolism, such as insulin resistance and glucose intolerance. Dysregulated glucose metabolism is an important manifestation of the disease and is the key to its pathogenesis. Therefore, studies involving evaluation of glucose metabolism in PCOS are of utmost importance. Very few studies have quantified hepatic glucose production directly in PCOS models using non-radioactive glucose tracers. In this study, we discuss step-by-step instructions for the quantification of the rate of hepatic glucose production in a PCOS mouse model by measuring M+2 enrichment of [6,6-2H2]glucose, a stable isotopic glucose tracer, via gas chromatography - mass spectrometry (GCMS). This procedure involves creation of stable isotopic glucose tracer solution, use of tail vein catheter placement and infusion of the glucose tracer in both fasting and glucose-rich states in the same mouse in tandem. The enrichment of [6,6-2H2]glucose is measured using pentaacetate derivative in GCMS. This technique can be applied to a wide variety of studies involving direct measurement of the rate of hepatic glucose production.

This study describes the direct measurement of hepatic glucose production in a polycystic ovary syndrome mouse model by using a stable isotopic glucose tracer via tail vein in both fasting and glucose-rich states in tandem.

Polycystic ovary syndrome (PCOS) is a common disease that results in disorders of glucose metabolism, such as insulin resistance and glucose intolerance. ...

Subcutaneous Dihydrotestosterone (DHT) Treatment: Mouse Model of Polycystic Ovary Syndrome (PCOS)

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