A Modified Technique for Inducing Polycystic Ovary Syndrome in Mice

Podsumowanie

This protocol introduces a technique for inducing polycystic ovary syndrome (PCOS) models in mice through controlled letrozole release using mini-pumps. Under adequate anesthesia, the mini-pump was implanted subcutaneously, and PCOS was successfully induced in the mice after a certain period of the mini-pump release.

Streszczenie

Polycystic ovary syndrome (PCOS) is one of the leading causes of infertility in women. Animal models are widely used to study the etiologic mechanisms of PCOS and for related drug development. Letrozole-induced mouse models replicate the metabolic and reproductive phenotypes of patients with PCOS. The traditional method of letrozole treatment in PCOS mice requires daily dosing over a certain period, which can be labor-intensive and cause significant stress to the mice. This study describes a simple and effective method for inducing PCOS in mice by implanting a controlled letrozole-releasing mini-pump. A mini-pump capable of stable, continuous release of a quantitative amount of letrozole was fabricated and implanted subcutaneously in mice under anesthesia. This study demonstrated that the mouse model successfully mimicked PCOS features after letrozole mini-pump implantation. The materials and equipment used in this study are readily available to most laboratories, requiring no special customization. Collectively, this article provides a unique, easy-to-perform method for inducing PCOS in mice.

Wprowadzenie

Polycystic ovary syndrome (PCOS) is one of the most common conditions among women of reproductive age¹. It affects up to 18% of women globally and is the leading cause of female infertility worldwide^2,3. PCOS is characterized by a series of interrelated reproductive abnormalities, including disturbed gonadotropin secretion, chronic anovulation, increased androgen production, and polycystic ovarian morphology⁴. In addition to gynecological disorders, PCOS also increases the risk of cardiovascular diseases^5,6

Protokół

All animal experimental protocols in this study were approved by Animal Ethics Committee of Fudan University. Female C57BL/6J mice, aged 4 weeks, were used here. The details of the animals, reagents, and equipment used in this study are listed in the Table of Materials.

1. Preparation of the mini-pump

1. Take the letrozole powder (50 mg) stored in the original packaging bottle, centrifuge the letrozole powder in the bottle at 300 x g for 5 s (at room temperature), then open the cap.
2. Slowly add 625 µL of DMSO to the letrozole powder with a pipette and mix well to ob....

Dyskusje

This report demonstrates a simple protocol for inducing PCOS in mice using easily accessible materials. The mouse PCOS model is essential for exploring PCOS mechanisms and screening drugs²⁶. Of the available methods for inducing PCOS models in mice, letrozole induction is one of the most commonly used. The use of letrozole can develop and maintain a hyperandrogenic condition by inhibiting the conversion of testosterone to estradiol or by increasing testosterone synthesis²⁷........

Materiały

Name	Company	Catalog Number	Comments
Syringe	Bofeng Biotech	BD300841	1ML (sterile)
Centrifugation tube	Biological Hope	1850-K	15ML (sterile)
Pipette	Eppendorf	3123000268-A	100μL-1000μL (sterile)
Pipette	TopPette	7010101008	10μL-100μL (sterile)
Letrozole powder	Sigma	L6545-50MG	Primary acting drugs (sterile)
PEG(Poly(ethylene glycol))	Solarbio	P8250	Used for dissolution (sterile)
Sterile Physiological Saline Solution	Biosharp	BL158A	Mini-pump storage
Osmotic Pumps	ALZET	1004	Letrozole storage and sustained release (sterile)
Dimethyl sulfoxide	Biosharp	BS087	Used for dissolution (sterile)
Small Animal Anesthesia Machine	RWD	R500IP	Used for anesthesia
Isoflurane	RWD	20071302	Used for anesthesia
Hemostats	Biosharp	BS-HF-S-125	Surgical instrument
Scissors	Biosharp	BS-SOR-S-100P	Surgical instrument
Needle holder	ShangHaiJZ	J32010	Surgical instrument
Forceps	RWD	F12028	Surgical instrument
Needle and the 4-0 absorbable suture	JINGHUAN	CR413	Surgical instrument
Povidone-iodine swabs	SingleLady	GB26368-2010	Skin disinfection
Depilatory cream	ZIKER BIOTECHNOLOGY	ZK-L2701	Depilation agent for laboratory animals
Nitrile Gloves	Biosharp	BC040-L	Used for aseptic operation
Sterile gauze	ZHENDE	BA69087	Used for wiping liquids
C57BL/6J Mice	Shanghai Model Organisms Center	N/A	Age: 4 weeks
Pet Eye Lubricant	BAITE	SHF021153	Used for mouse eye lubricant
Meloxicam Injection	MEIDAJIA	R21064-21	Used for mouse analgesia (sterile)