Profiles of Steroid Hormones in Canine X-Linked Muscular Dystrophy via Stable Isotope Dilution LC-MS/MS

• Published in: PloS one Vol. 10; no. 5; p. e0126585
• Main Authors: Martins-Júnior, Helio A, Simas, Rosineide C, Brolio, Marina P, Ferreira, Christina R, Perecin, Felipe, Nogueira, Guilherme de P, Miglino, Maria A, Martins, Daniele S, Eberlin, Marcos N, Ambrósio, Carlos E
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 26.05.2015; Public Library of Science (PLoS)
• Subjects: 17β-Estradiol; Animals; Chromatography; Chromatography, Liquid; Cortisol; Development and progression; Diagnosis; Dilution; Discriminant Analysis; Dog diseases; Dog Diseases - blood; Dogs; Duchenne's muscular dystrophy; Dystrophy; Estrus cycle; Female; Genetic Diseases, X-Linked - blood; Health aspects; Hormones; Hormones - blood; Identification and classification; Isotope Labeling - methods; Liquid chromatography; Mass spectrometry; Mass spectroscopy; Multivariate statistical analysis; Muscular dystrophy; Muscular Dystrophy, Animal - blood; Phase transitions; Principal Component Analysis; Progesterone; Quality of life; Reproducibility of Results; Rodents; Sex hormones; Statistical analysis; Steroid hormones; Steroids - blood; Studies; Tandem Mass Spectrometry - methods; Testosterone
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0126585

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.