title: Profiles of Steroid Hormones in Canine X-Linked Muscular Dystrophy via Stable Isotope Dilution LC-MS/MS.
abstract: Golden retriever muscular dystrophy (GRMD) provides the best animal model for characterizing the disease progress of the human disorder, Duchenne muscular dystrophy (DMD). The purpose of this study was to determine steroid hormone concentration profiles in healthy golden retriever dogs (control group - CtGR) versus GRMD-gene carrier (CaGR) and affected female dogs (AfCR). Therefore, a sensitive and specific analytical method was developed and validated to determine the estradiol, progesterone, cortisol, and testosterone levels in the canine serum by isotope dilution liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). To more accurately understand the dynamic nature of the serum steroid profile, the fluctuating levels of these four steroid hormones over the estrous cycle were compared across the three experimental groups using a multivariate statistical analysis. The concentration profiles of estradiol, cortisol, progesterone, and testosterone revealed a characteristic pattern for each studied group at each specific estrous phase. Additionally, several important changes in the serum concentrations of cortisol and estradiol in the CaGR and AfCR groups seem to be correlated with the status and progression of the muscular dystrophy. A comprehensive and quantitative monitoring of steroid profiles throughout the estrous cycle of normal and GRMD dogs were achieved. Significant differences in these profiles were observed between GRMD and healthy animals, most notably for estradiol. These findings contribute to a better understanding of both dog reproduction and the muscular dystrophy pathology. Our data open new venues for hormonal behavior studies in dystrophinopathies and that may affect the quality of life of DMD patients.
authors.affiliation: ThoMSon Mass Spectrometry Laboratory-Institute of Chemistry, University of Campinas (UNICAMP), Campinas, São Paulo, Brazil; AB SCIEX of Brazil, São Paulo, São Paulo, Brazil.
authors.forename: Helio A
authors.initials: HA
authors.lastname: Martins-Júnior
authors.affiliation: ThoMSon Mass Spectrometry Laboratory-Institute of Chemistry, University of Campinas (UNICAMP), Campinas, São Paulo, Brazil; DAPSA, Faculty of Veterinary Medicine, São Paulo State University-UNESP, Araçatuba, São Paulo, Brazil.
authors.forename: Rosineide C
authors.initials: RC
authors.lastname: Simas
authors.affiliation: Faculty of Animal Sciences and Food Engineering-FZEA, University of São Paulo-USP, Pirassununga, São Paulo, Brazil.
authors.forename: Marina P
authors.initials: MP
authors.lastname: Brolio
authors.affiliation: ThoMSon Mass Spectrometry Laboratory-Institute of Chemistry, University of Campinas (UNICAMP), Campinas, São Paulo, Brazil; Department of Chemistry, Purdue University, West Lafayette, Indiana, United States of America.
authors.forename: Christina R
authors.initials: CR
authors.lastname: Ferreira
authors.affiliation: Faculty of Animal Sciences and Food Engineering-FZEA, University of São Paulo-USP, Pirassununga, São Paulo, Brazil.
authors.forename: Felipe
authors.initials: F
authors.lastname: Perecin
authors.affiliation: DAPSA, Faculty of Veterinary Medicine, São Paulo State University-UNESP, Araçatuba, São Paulo, Brazil.
authors.forename: Guilherme de P
authors.initials: Gde P
authors.lastname: Nogueira
authors.affiliation: Faculty of Veterinary Medicine and Animal Science-FMVZ, University of São Paulo-USP, São Paulo, São Paulo, Brazil.
authors.forename: Maria A
authors.initials: MA
authors.lastname: Miglino
authors.affiliation: Faculty of Animal Sciences and Food Engineering-FZEA, University of São Paulo-USP, Pirassununga, São Paulo, Brazil.
authors.forename: Daniele S
authors.initials: DS
authors.lastname: Martins
authors.affiliation: ThoMSon Mass Spectrometry Laboratory-Institute of Chemistry, University of Campinas (UNICAMP), Campinas, São Paulo, Brazil.
authors.forename: Marcos N
authors.initials: MN
authors.lastname: Eberlin
authors.affiliation: Faculty of Animal Sciences and Food Engineering-FZEA, University of São Paulo-USP, Pirassununga, São Paulo, Brazil.
authors.forename: Carlos E
authors.initials: CE
authors.lastname: Ambrósio
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid; D016002:Discriminant Analysis; D004283:Dog Diseases / Q000097:blood*; D004285:Dogs; D005260:Female; D040181:Genetic Diseases, X-Linked / Q000097:blood*; D006728:Hormones / Q000097:blood*; D007553:Isotope Labeling / Q000379:methods*; D009137:Muscular Dystrophy, Animal / Q000097:blood*; D025341:Principal Component Analysis; D015203:Reproducibility of Results; D013256:Steroids / Q000097:blood*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D006728:Hormones; D013256:Steroids