[Show abstract][Hide abstract] ABSTRACT: The present study describes the seminal plasma proteome of Bos indicus bulls. Fifty-six, 24-month old Australian Brahman sires were evaluated and subjected to electroejaculation. Seminal plasma proteins were separated by 2-D SDS-PAGE and identified by mass spectrometry. The percentage of progressively motile and morphologically normal sperm of the bulls were 70.4±2.3 and 64±3.2%, respectively. A total of 108 spots were identified in the 2-D maps, corresponding to 46 proteins. Binder of sperm proteins accounted for 55.8% of all spots detected in the maps and spermadhesins comprised the second most abundant constituents. Other proteins of the Bos indicus seminal plasma include clusterin, albumin, transferrin, metalloproteinase inhibitor 2, osteopontin, epididymal secretory protein E1, apolipoprotein A-1, heat shock 70kDa protein, glutathione peroxidase 3, cathelicidins, alpha-enolase, tripeptidyl-peptidase 1, zinc-alpha-2-glycoprotein, plasma serine protease inhibitor, beta 2-microglobulin, proteasome subunit beta type-4, actin, cathepsins, nucleobinding-1, protein S100-A9, hemoglobin subunit alpha, cadherin-1, angiogenin-1, fibrinogen alpha and beta chain, ephirin-A1, protein DJ-1, serpin A3-7, alpha-2-macroglobulin, annexin A1, complement factor B, polymeric immunoglobulin receptor, seminal ribonuclease, ribonuclease-4, prostaglandin-H2 d-isomerase, platelet-activating factor acetylhydrolase, and phosphoglycerate kinase 1. In conclusion, this work uniquely portrays the Bos indicus seminal fluid proteome, based on samples from a large set of animals representing the Brahman cattle of the tropical Northern Australia. Based on putative biochemical attributes, seminal proteins act during sperm maturation, protection, capacitation and fertilization.
[Show abstract][Hide abstract] ABSTRACT: Recently, comprehensive studies were conducted regarding the reproductive development, age at puberty, spermatogenesis and the protein profile in the seminal plasma of Santa Ines rams. Despite the abundant information obtained from these studies regarding these tropically adapted rams in Brazil, it is still unclear how sperm parameters relate to the expression of molecular components of the reproductive tract. In this regard, the present study was conducted to determine if sperm parameters were empirically associated with the seminal plasma proteins described in two-dimensional electrophoresis maps. Seminal plasma proteins were separated by two-dimensional electrophoresis and the respective maps electronically analyzed. Protein spots associated with higher or lower sperm motility were then identified by tandem mass spectrometry. Based on sperm motility, the ejaculates were divided into two groups: those containing up to 80% motile cells (G1; n = 11) and those with more than 80% motile sperm (G2; n = 10). On average, 236 spots per gel were detected. Eleven spots were significantly more intense in gels from animals with ejaculates characterized by higher semen motility scores (G2). The intensity of three other spots was higher in gels from the G1 group. All spots differentially expressed for G1 and G2, were present in at least 90% of the gels. From the 13 spots differentially expressed in G1 and G2, four spots were identified by tandem mass spectrometry. Spots expressed with more intensity in the ejaculates with higher sperm motility (G2) were identified as arylsulfatase A and zinc-alpha-2-glycoprotein. On the other hand, two spots associated with G1 were identified as ram seminal vesicles protein 22 kDa (RSVP-22) and bodhesin-2. Knowledge of these identities represents a crucial step toward the comprehension of how specific seminal plasma proteins are related to sperm motility.
Small Ruminant Research 01/2013; 109(2–3):94-100. · 1.12 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The present study is focused on the proteome of reproductive tract fluids from tropically-adapted Santa Ines rams. Seminal plasma, cauda epididymal (CEF) and vesicular gland fluid (VGF) proteins were analyzed by 2-D electrophoresis and mass spectrometry. Seminal plasma maps contained 302 ± 16 spots, within the 4-7 pH range. From these maps, 73 spots were identified, corresponding to 41 proteins. Ram Seminal Vesicle Proteins (RSVP) 14 and 22kDa and bodhesins 1 and 2 represented the most abundant seminal components. Other seminal proteins included clusterin, angiotensin-converting enzyme, matrix metalloproteinase-2, tissue-inhibitor of metalloproteinase-2, plasma glutamate carboxypeptidase, albumin, lactoferrin, alpha enolase, peroxiredoxin, leucine aminopeptidase, β-galactosidase, among others. Later, seminal plasma gels were run within narrow pH intervals (3.9-5.1; 4.7-5.9; 5.5-6.7), allowing the additional identification of 21 proteins not detected in 4-7 pH maps. Major proteins of CEF and VGF were albumin and transferrin, and RSVPs, respectively. Western blots confirmed that RSVPs were mainly present in VGF while bodhesins, in VGF and CEF. Based on RT-PCR, RSVP and bodhesin genes were primarily expressed in the vesicular glands. In summary, the reproductive tract fluids of Brazilian hairy rams contain several categories of proteins, with potential roles in sperm protection, capacitation, acrosome reaction and sperm-oocyte interaction.
Journal of proteomics 05/2012; 75(14):4436-56. · 5.07 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy.
[Show abstract][Hide abstract] ABSTRACT: Santa Ines is a native Brazilian breed of hair sheep, representing a genotype developed in and adapted to tropical regions. Seminal plasma of the rams was obtained, subjected to two-dimensional electrophoresis and the major spots identified by tandem mass spectrometry. There were 143 spots consistently present on seminal plasma maps. The most abundant spots represented two families, binder of sperm proteins (BSP) and spermadhesins. The proteins included ram seminal vesicles proteins (RSVP) 14kDa and 22kDa, and bodhesin 1 and 2. This is the first report of identification of seminal plasma proteins of tropically-adapted sheep. These proteins are involved in several aspects of reproductive function, including sperm capacitation, sperm survival in the oviduct and gamete interaction in mammals.