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Identification of diabetes- and obesity-associated proteomic changes in human spermatozoa by difference gel electrophoresis
Authors:Thomas M Kriegel  Falk Heidenreich  Karina Kettner  Theresia Pursche  Bernard Hoflack  Sonja Grunewald  Kerstin Poenicke  Hans-Juergen Glander  Uwe Paasch
Affiliation:1. Dresden University of Technology, Carl Gustav Carus Medical Faculty, Institute of Physiological Chemistry, 01307 Dresden, Germany;2. Dresden University of Technology, Biotechnology Center, Proteomics Group, 01307 Dresden, Germany;3. University of Leipzig, Department of Dermatology, Training Center of the European Academy of Andrology, 04103 Leipzig, Germany;4. TMK and FH contributed equally to this work.;1. The Department of Obstetrics and Gynecology, Meir Medical Center, Kfar-Saba, Israel;2. Sakler School of Medicine, School of Public Health, Tel Aviv University, Tel Aviv, Israel;3. Assuta Medical Center, Tel Aviv, Israel;1. Departamento de Reproducción Animal, SGIT-INIA, Madrid, Spain;2. Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Brasilia DF, Brazil;3. Departamento de Producción Animal, IMIDA, Murcia, Spain;1. Andrology and IVF Laboratory, Department of Surgery (Urology), University of Utah, Salt Lake City, Utah;2. Department of Obstetrics and Gynecology, University of Utah, Salt Lake City, Utah;3. Department of Human Genetics, University of Utah, Salt Lake City, Utah;1. Department of Surgery, Division of Urology, Human Reproduction Section, São Paulo Federal University, São Paulo, Brazil;2. Fleury Group, São Paulo, Brazil;3. São Paulo Hospital, São Paulo, Brazil;1. Department of Neuroendocrinology, National Institute for Research in Reproductive Health (ICMR), Jehangir Merwanji Street, Parel, Mumbai 400012, India;2. National Centre for Laboratory Animal Sciences (NCLAS), National Institute of Nutrition, Jamai-Osmania PO Hyderabad, 500 007, India;1. IANENTRO Fertility Center, Thessaloniki, Greece;2. Centre for Reproductive Medicine, Department of Obstetrics and Gynecology, Ioannina University School of Medicine, Ioannina, Greece;3. IVF Centers Prof. Zech, Bregenz, Austria;4. Centre Hospitalier Inter Regional Cavell (CHIREC), Braine l‘Alleud – Bruxelles, Belgium;5. 4th Departments of Obstetrics and Gynecology, Aristotle University of Thessaloniki, Greece
Abstract:Difference gel electrophoresis (DIGE) of fluorescently labelled human sperm proteins was used to identify diabetes- and obesity-associated changes of the sperm proteome. Semen samples from type 1 diabetics, non-diabetic obese individuals and a reference group of clinically healthy fertile donors were evaluated in a comparative study. The adaptation of a general protein extraction procedure to the solubilization of proteins from isolated progressively motile human spermatozoa resulted in the detection of approximately 2700 fluorescent protein spots in the DIGE images. Comparison of the patients’ sperm proteomes with those of the reference group allowed the identification of 20 spots containing proteins that were present in the sperm lysates at significantly increased or decreased concentrations. In detail, eight of these spots were apparently related to type 1 diabetes while 12 spots were apparently related to obesity. Tryptic digestion of the spot proteins and mass spectrometric analysis of the corresponding peptides identified seven sperm proteins apparently associated with type 1 diabetes and nine sperm proteins apparently associated with obesity, three of which existing in multiple molecular forms. The established proteomic approach is expected to function as a non-invasive experimental tool in the diagnosis of male infertility and in monitoring any fertility-restoring therapy.
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