WCRB2014 POSTER PRESENTATIONS (1) (335 abstracts)
Proteomic analysis of human testicular interstitial fluid reflects disordered spermatogenesis in Klinefelter’s syndrome
Robert I McLachlan 1 , Andrew N Stephens 1 , Adam Rainczuk 1 , Caroline Foo 1 , Mark R Condina 2 , Tomomoto Ishikawa 3 , Wolfgang Weidner 4 & Peter G Stanton 1
1MIMIR-PHI Institute, Clayton, Victoria, Australia; 2Bruker Pty. Ltd, Melbourne, Victoria, Australia; 3Ishikawa Hospital, Himeji, Japan; 4Justus Liebig University, Giessen, Germany.
Introduction: Primary spermatogenic failure is the commonest cause of male infertility. Changes in Sertoli and germ cell function may be reflected in the testicular interstitial fluid (TIF) proteome and its analysis may provide insights into the pathophysiology. Microdissection testicular sperm extraction (microTESE) provides sperm for ICSI in ~50% of non-obstructive azoospermia (NOA) cases. As the first step in constructing a diagnostic approach for broader application in NOA, we chose to examine the TIF proteome of Klinefelter’s men with severely impaired spermatogenesis.
Materials and methods: We pooled TIF from Klinefelter’s syndrome (KS) men with a predominantly Sertoli cell-only histological phenotype, but in whom spermatids were found at microTESE, and compared their TIF proteomic profile with that of control (obstructed) cases (n=3/group). Low-abundant proteins were isolated from each TIF pool using BioRad Proteominer beads, and labelled using ICPL chemistry to quantify protein expression differences by mass spectrometry (MS). Samples were combined, digested with trypsin, and peptides pre-fractionated by nano-reversed phase HPLC, prior to identification using MALDI tandem MS and ESI-LCMS/MS.
Results and discussion: 1248 proteins were characterized; 59 were down regulated greater than two-fold (including known Sertoli cell- or germ cell-specific proteins) and 70 up regulated greater than two-fold (these tended to reflect altered intracellular signalling or regulation of somatic cell origin). We conclude that Sertoli and germ cells proteins can readily be recognised in TIF, and that differences in their expression provides a promising basis upon which to identify sub-phenotypes of spermatogenic failure and a means to predict the recovery of sperm at TESE.
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ISSN 2052-1472 (online)
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