Department of Animal Medicine, Veterinary Faculty, Federal University of Rio Grande do Sul, Porto Alegre, RS, Brazil.
Theriogenology. 2011 Sep 1;76(4):765-71. doi: 10.1016/j.theriogenology.2011.04.010. Epub 2011 May 23.
The objective was to evaluate protein profiles of equine seminal plasma using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and to determine whether any of these proteins were related to semen freezability. Seminal plasma was collected from 10 stallions, of high and low semen freezability, housed at the State Stud of Lower Saxony, and routinely used in AI programs. Twenty-five protein spots were identified from the two-dimensional gel (12%), seven of which were present in all samples (all proteins were identified by MALDI-MS). Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) has been used to generate ion images of samples in one or more mass-to-charge (m/z) values, providing the capability of mapping specific molecules to two-dimensional coordinates of the original sample. Of the 25 proteins identified, two spots had greater relative content (P < 0.05) in seminal plasma samples collected from stallions with high semen freezability: spot 5 (80-85 kDa, isoelectric point [pI] 7.54), identified as CRISP-3; and spot 45 (18.2 kDa, pI 5.0-5.2), identified as HSP-2. Conversely, protein content was greater (P < 0.05) in seminal plasma samples from stallions with low semen freezability: spot 7 (75.4 kDa, pI 6.9-7.4), identified as lactoferrin; spot 15 (26.7 kDa, pI 5.51), identified as kallikrein; spot 25 (25 kDa, pI 7.54), identified as CRISP-3; and spot 35 (13.9 kDa, pI 3.8-4.2), identified as HSP-1. In conclusion, there were differences in the seminal plasma protein profile from stallions with high and low semen freezability. Furthermore, CRISP-3 and HSP-2 were potential seminal plasma markers of high semen freezability.
目的是使用二维聚丙烯酰胺凝胶电泳（2D-PAGE）评估马精液的蛋白质图谱，并确定这些蛋白质中是否有与精液冷冻能力相关的蛋白质。精液是从居住在下萨克森州国家种马场、且常规用于人工授精计划的 10 匹高和低精液冷冻能力的种马中收集的。从二维凝胶中鉴定出 25 个蛋白质斑点（12%），其中 7 个存在于所有样本中（所有蛋白质均通过 MALDI-MS 鉴定）。基质辅助激光解吸/电离质谱（MALDI-MS）已被用于在一个或多个质荷比（m/z）值下生成样品的离子图像，从而提供将特定分子映射到原始样品二维坐标的能力。在鉴定出的 25 种蛋白质中，有 2 个斑点在高精液冷冻能力的种马精液中具有更高的相对含量（P<0.05）：斑点 5（80-85 kDa，等电点 [pI] 7.54），鉴定为 CRISP-3；斑点 45（18.2 kDa，pI 5.0-5.2），鉴定为 HSP-2。相反，在低精液冷冻能力的种马精液中蛋白质含量更高（P<0.05）：斑点 7（75.4 kDa，pI 6.9-7.4），鉴定为乳铁蛋白；斑点 15（26.7 kDa，pI 5.51），鉴定为激肽释放酶；斑点 25（25 kDa，pI 7.54），鉴定为 CRISP-3；斑点 35（13.9 kDa，pI 3.8-4.2），鉴定为 HSP-1。总之，高和低精液冷冻能力的种马精液的蛋白质图谱存在差异。此外，CRISP-3 和 HSP-2 可能是高精液冷冻能力的潜在精液蛋白质标志物。
