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Abstract

Introduction

Protocol

Representative Results

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Materials

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Developmental Biology

A Hyperandrogenic Mouse Model to Study Polycystic Ovary Syndrome

Published: October 2nd, 2018

DOI:

10.3791/58379

1Department of Pediatrics, Johns Hopkins University School of Medicine, 2Department of Health, Beijing Military General Hospital, 3Southern Medical University, 4Department of Molecular and Cellular Physiology, Johns Hopkins University School of Medicine, 5Department of Gynecology and Obstetrics, Johns Hopkins University School of Medicine

We describe the development of a lean PCOS-like mouse model with dihydrotestosterone pellet to study the pathophysiology of PCOS and the offspring from these PCOS-like dams.

Hyperandrogenemia plays a critical role in reproductive and metabolic function in females and is the hallmark of polycystic ovary syndrome. Developing a lean PCOS-like mouse model that mimics women with PCOS is clinically meaningful. In this protocol, we describe such a model. By inserting a 4 mm length of DHT (dihydrotestosterone) crystal powder pellet (total length of pellet is 8 mm), and replacing it monthly, we are able to produce a PCOS-like mouse model with serum DHT levels 2 fold higher than mice not implanted with DHT (no-DHT). We observed reproductive and metabolic dysfunction without changing body weight and body composition. While exhibiting a high degree of infertility, a small subset of these PCOS-like female mice can get pregnant and their offspring show delayed puberty and increased testosterone as adults. This PCOS-like lean mouse model is a useful tool to study the pathophysiology of PCOS and the offspring from these PCOS-like dams.

Hyperandrogenism is the hallmark of polycystic ovary syndrome (PCOS) according to NIH criteria and of the Androgen Excess and PCOS (AE-PCOS) Society. Women with PCOS have difficulty getting pregnant and have increased risk of pregnancy complications1. Even if they get pregnant, their female offspring have an adverse health outcomes2,3. Animal models have been developed using various strategies4,5,6,7,8,

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Here, we present detailed protocols for DHT pellet preparation and insertion, and for reproductive and metabolic testing. The mice used in this study were a mixed background (C57/B6, CD1, 129Sv) and were maintained with food and water ad libitum in a 14/10 h light/dark cycle at 24 °C in the Broadway Research Building animal facility at the Johns Hopkins University School of Medicine. All procedures were approved by the Johns Hopkins University Animal Care and Use Committee.

1. Crea.......

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Serum DHT levels and Glucose tolerance test

DHT levels were measured from collected serum by both ELISA and by LC-MS according to protocol 1.24–1.25, and 2.9, 3.0. The DHT absolute values are different between mass spectrometry and ELISA, however, the relative fold (around 2-fold) of DHT vs no-DHT insertion is similar from both assays and across experiments15,23,.......

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Hyperandrogenism is a key feature of PCOS. The serum DHT levels (two fold higher in DHT mice than in no-DHT mice) used in this protocol are lower than those reported by other investigators in previous studies and are calibrated to proportionally mimic women with PCOS5,19,20,21. Unlike other models, this 2-fold DHT model does not alter the body weight and whole body composition compared with no-.......

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This work was supported by the National Institutes of Health (Grants R00-HD068130 to S.W.) and the Baltimore Diabetes Research Center: Pilots and Feasibility Grant (to S.W.).

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Name Company Catalog Number Comments
Crystalline 5α-DHT powder   Sigma-Aldrich A8380-1G
Dow Corning Silastic tubing Fisher Scientific 11-189-15D 0.04in/1mm inner diameter x0.085in/2.15mm outer diameter
Medical adhesive silicone  Factor II, InC.  A-100
Goggles, lab coats, gloves and masks.
 10 µL pipette tips without filter USA Scientific 11113700
Microscope slide for smear Fisher Scientific 12-550-003
Diff Quik for staining cells Fisher Scientific NC9979740
  Lancet Fisher Scientific NC9416572
3 mL Syring  Becton, Dickinson and Company (BD), 30985
 attached needle: 20G BD 305176
 Ruler: any length than 10cm with milimeter scale. 
Xylazine  Vet one AnnSeA LA, MWI, Boise NDC13985-704-10 100mg/ml
Ketamine Hydrochloride Hospira, Inc NDC 0409-2051-05 100mg/ml
 Surgical staple  AutoClip® System, Fine Science Tool 12020-00
 Insulin syringe BD 329461 1/2 CC, low dose U-100 insulin syringe
 Trochar  Innovative Research of America MP-182
Microscope Carl Zeiss Primo Star 415500-0010-001 Germany
Ear punch Fisher Scientific 13-812-201
Testosterone rat/mouse ELISA kit IBL B79174
DHT ELISA kit Alpha Diagnostic International 1940
One touch ultra glucometer Life Scan, Inc.
One touch ultra test stripes Life Scan, Inc.
Eppendorf tube Fisher Scientific 05-402-18
Razor blade Fisher Scientific 12-640
Clidox Fisher Scientific NC0089321
surgical underpad Fisher Scientific 50587953 Supplier Diversity Partner
Manufacturer:  Andwin Scientific 56616018
Betadine Antiseptic Solution Walgreens
3M Vetbond (n-butyl cyanoacrylate) 3M Science. Applied to Life
Animal tattoo ink paste Ketchum manufacturing Inc. Brockville, Ontario, Canada
Scale Ohaus Corporation  HH120D Pine Brook, NJ
Electronic digital caliper NEIKO Tools USA 01407A available from Amazon

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