Identification of diabetes- and obesity-associated proteomic changes in human spermatozoa by difference gel electrophoresis

Kriegel, Thomas M; Heidenreich, Falk; Kettner, Karina; Pursche, Theresia; Hoflack, Bernard; Grunewald, Sonja; Poenicke, Kerstin; Glander, Hans-Juergen; Paasch, Uwe

doi:10.1016/j.rbmo.2009.07.001

« Previous Next »Reproductive BioMedicine Online
Volume 19, Issue 5 , Pages 660-670, November 2009

Identification of diabetes- and obesity-associated proteomic changes in human spermatozoa by difference gel electrophoresis

Thomas M Kriegel
- Dresden University of Technology, Carl Gustav Carus Medical Faculty, Institute of Physiological Chemistry, 01307 Dresden, Germany
- TMK and FH contributed equally to this work.
Falk Heidenreich
- Dresden University of Technology, Carl Gustav Carus Medical Faculty, Institute of Physiological Chemistry, 01307 Dresden, Germany
- TMK and FH contributed equally to this work.
Karina Kettner
- Dresden University of Technology, Carl Gustav Carus Medical Faculty, Institute of Physiological Chemistry, 01307 Dresden, Germany
Theresia Pursche
- Dresden University of Technology, Biotechnology Center, Proteomics Group, 01307 Dresden, Germany
Bernard Hoflack
- Dresden University of Technology, Biotechnology Center, Proteomics Group, 01307 Dresden, Germany
Sonja Grunewald
- University of Leipzig, Department of Dermatology, Training Center of the European Academy of Andrology, 04103 Leipzig, Germany
Kerstin Poenicke
- University of Leipzig, Department of Dermatology, Training Center of the European Academy of Andrology, 04103 Leipzig, Germany
Hans-Juergen Glander
- University of Leipzig, Department of Dermatology, Training Center of the European Academy of Andrology, 04103 Leipzig, Germany
Uwe Paasch
- University of Leipzig, Department of Dermatology, Training Center of the European Academy of Andrology, 04103 Leipzig, Germany
- Correspondence:

Received 14 January 2009; received in revised form 12 February 2009; accepted 6 July 2009. published online 02 August 2010.

Declaration: The authors report no financial or commercial conflicts of interest.

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.

Keywords: DIGE, human spermatozoa, obesity, proteomics, type 1 diabetes

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Professor Paasch completed his studies in medicine in 1995 at the University of Leipzig, Germany. Since 2001, he has been an assistant Professor in Andrology at the Department of Andrology at the University of Leipzig, which became a certified Training Centre of the European Academy of Andrology in 1996. Professor Paasch was among the first to describe annexin binding to human spermatozoa and to develop molecular selection strategies. Recently, he and his team investigated apoptotic signal transduction, lipid changes at the membrane, and related sperm dysfunction.Professor Uwe Paasch