DOI: 10.5061/dryad.wwpzgmsr7
title: Type I fiber decrease and ectopic fat accumulation in skeletal muscle from women with PCOS

description: Background: Polycystic ovary syndrome’s (PCOS) main feature is
 hyperandrogenism, which is linked to a higher risk of metabolic disorders
 in women. Gene expression analyses in adipose tissue and skeletal muscle
 reveal dysregulated metabolic pathways in women with PCOS, but these
 differences do not necessarily lead to changes in protein levels and
 biological function. Methods: To advance our understanding of the
 molecular alterations in PCOS, we performed global proteomic and
 phosphorylation site analysis using tandem mass spectrometry. Adipose
 tissue and skeletal muscle were collected at baseline from 10 women with
 and without PCOS, and in women with PCOS after 5 weeks of treatment with
 electrical stimulation. Results: Perilipin-1, a protein that typically
 coats the surface of lipid droplets in adipocytes, was increased whereas
 proteins involved in muscle contraction and type I muscle fiber function
 were downregulated in PCOS muscle. Proteins in the thick and thin
 filaments had many altered phosphorylation sites, indicating differences
 in protein activity and function. The upregulated proteins in muscle post
 treatment were enriched in pathways involved in extracellular matrix
 organization and wound healing, which may reflect a protective adaptation
 to repeated contractions and tissue damage due to needling. A similar,
 albeit less pronounced, upregulation in extracellular matrix organization
 pathways was also seen in adipose tissue. Conclusions: Our results suggest
 that hyperandrogenic women with PCOS have higher levels of
 extramyocellular lipids and fewer oxidative insulin-sensitive type I
 muscle fibers. These could be key factors leading insulin resistance in
 PCOS muscle while electric stimulation-induced tissue remodeling may be
 protective.
description: Women with polycystic ovary syndrome (PCOS) were diagnosed
 according to the Rotterdam criteria. Subcutaneous adipose tissue and
 skeletal muscle biopsies from women with PCOS and controls were included
 in the proteomics analysis. Immunfluorescence was used to quantify
 perilipin-1 and slow type I myosin fibers in skeletal muscle. Picrosirius
 Red staining was used to quantify fibrillar collagen in adipose and muscle
 tissue. 3-week-old wild-type (wt) female mice on
 C57Bl/6J background were purchased from Janvier Labs (C57BL/6NRj, Le
 Genest-Saint-Isle, France). Female skeletal muscle androgen receptor
 knockout mice (SkMARKO) were generated by crossing ARflox mice with
 B6;C3-Tg(ACTA1-rtTA,tetO-cre)102Monk/J (HSA-rtTA/TRE-Cre) mice. To induce
 Cre recombinase expression, at 3 weeks of age, SkMARKO mice were given a
 diet containing 200 mg/kg doxycycline (Specialty Feeds SF11-059) for the
 entire duration of the experiment. At 4 weeks of age, wt and SkMARKO
 female mice received a subcutaneous silastic implant containing 5-10 mg
 DHT (5α-Androstan-17β-ol-3-one, A8380, Sigma-Aldrich, St. Louis, USA) or
 empty implant (n=5-8/group). A subset of DHT-exposed wt mice received a
 slow-release flutamide pellet (n=5, 25 mg flutamide/pellet, 90-day
 release, Innovative Research of America, Sarasota). At 15-17 weeks of age,
 the mice were euthanized and gastrocnemius muscle tissue was dissected and
 snap-frozen. Western blot was used to quantify slow type I myosin fibers
 in skeletal muscle.
description: # Type I fiber decrease and ectopic fat accumulation in skeletal muscle
 from women with PCOS
 [https://doi.org/10.5061/dryad.wwpzgmsr7](https://doi.org/10.5061/dryad.wwpzgmsr7) This data set contains the quantification of human muscle fiber cross-sectional area and percentage of MYH7 positive type I fibers (FCSA_PCOS_vs_Ctrl.xlsx) and perilipin-1 staining (Perilipin_PCOS_vs_Ctrl.xlsx). It also contains the quantified fibrillar collagen content in adipose tissue from women with polycystic ovary syndrome (PCOS) and controls (Collagen_quantification.xlsx), and collagen content in adipose and skeletal muscle tissue in women with PCOS after 5 weeks of electrical stimulation treatment (Collagen adipose tissue treatment)(Collagen muscle tissue treatment). I contains the quantified MYH7 protein expression in skeletal muscle from wt and SkMARKO female mice exposed to DHT prepubertaly, to induce a PCOS-like phenotype (MYH7_mouse.xlsx). ## Description of the data and file structure **FCSA_PCOS_vs_Ctrl.xlsx** PCOS= polycystic ovary syndrome Ctrl= control FCSA: fiber cross-sectional area  content: Human muscle fiber cross-sectional area in seven controls and five women with PCOS. The file contains individual values of all analyzed muscle fibers from each subject. Percentage of type 1 muscle fibers in PCOS vs controls: the percentage of type 1 muscle fibers in four women with PCOS and four controls. **Perilipin_PCOS_vs_Ctrl.xlsx** PCOS= polycystic ovary syndrome units: arbitrary units of mean and standard deviation (SD) of staining intensity Content: quantification of Perilipin staining in skeletal muscle from women with PCOS and controls. **Collagen_quantification.xlsx** PCOS= polycystic ovary syndrome Collagen adipose tissue PCOS vs: Collagen content in adipose tissue from seven women with polycystic ovary syndrome (PCOS) and four controls.  units: data are presented as mean value of arbitrary units per individual. Collagen muscle tissue treatment: Collagen content in skeletal muscle tissue in four women with PCOS before (baseline) and after 5 weeks of electrical stimulation (treatment). units: data are presented as mean arbitrary unit value per individual. Collagen adipose tissue treatment: Collagen content in adipose tissue in six women with PCOS before (baseline) and after 5 weeks of electrical stimulation (treatment). units: data are presented as mean arbitrary unit value per individual. **MYH7_mouse.xlsx** Content: MYH7 protein expression in gastrocnemius muscle from wt mice exposed to DHT or DHT + flutamide units: arbitrary units related to GAPDH expression is presented in the paper Treatment: control, DHT exposed, or flutamide exposed mice Geno.= genotype. All mice are wildtypes (WT) DHT= dihydrotestosterone Myosin: arbitrary units of myosin band on the western blot membrane GAPDH: arbitrary units of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) band on the western blot membrane Myosin/GAPDH: ratio between myosin and GAPDH arbitrary unit expression ## Sharing/Access information Links to other publicly accessible locations of the data: The proteomics data have been deposited to the ProteomeXchange Consortium via the Proteomics Identifications (PRIDE) (RRID:SCR_003411) partner repository with the dataset identifier PXD025358. ## Code/Software The protein expression analysis is published at [https://github.com/GustawEriksson/PCOS](https://github.com/GustawEriksson/PCOS)