Submitted to: Poultry Science Association Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: April 15, 2009
Publication Date: July 20, 2009
Citation: Long, J.A. 2009. Proteomic Assessment of Poultry Spermatozoa. Poultry Science 88 (Suppl. 1) Abstract 57 p.18 (CD-Rom). Technical Abstract: Fully characterizing the protein composition of spermatozoa is the first step in utilizing proteomics to delineate the function of sperm proteins. To date, sperm proteome maps have been partially developed for the human, mouse, rat, bull and several invertebrates. Here we report the first proteomic analysis of turkey and rooster spermatozoa, using MALDI-TOF and LC-MS/MS. Semen was centrifuged through a discontinuous Accudenz gradient to remove seminal plasma. Protein was extracted from isolated sperm cells and the soluble fraction separated by 2-D SDS-PAGE (pI 5-8). Excised spots were digested with trypsin and prepared for MALDI-TOF analysis. Proteins were identified from Peptide Mass Fingerprints using the MASCOT search engine (Matrix Science). Samples yielding non-significant Mowse probability scores were subjected to LC-MS/MS analysis. When necessary, homology searches were performed for unnamed protein products via BLAST searching. A total of 94 and 36 proteins were identified from turkey and rooster spermatozoa, respectively. All proteins identifications were limited to Gallus gallus and/or Meleagris gallopavo. For turkey sperm, 9 hypothetical proteins (6 matching to chicken chromosome open reading frames via BLAST search) were identified, while another16 were predicted proteins of unknown function. For rooster sperm, 5 hypothetical proteins (2 matching to chicken chromosome open reading frames) were identified, and an additional predicted protein of unknown function was positively identified. Identified proteins were associated with the acrosome (pro-acrosin), mitochondria (enolase I, voltage-dependent anion channel 2, creatine kinase), and flagellum (capping protein, dynein, tektins 1-5). Several chaperone (heat shock protein 70) and calcium-binding (EF-hand protein) proteins also were identified. Three proteins not previously found in sperm were identified: dihydropyrimidinase, mitofilin and mitochondrial tri-functional protein. While the latter 2 mitochondrial proteins most likely exist in sperm from other species, the discovery of dihydropyrimidinase as a predominant soluble protein in poultry sperm poses interesting functional implications.