Department of Gametes and Embryo Biology, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Tuwima 10, 10-714 Olsztyn, Poland.
Fish Physiol Biochem. 2011 Sep;37(3):447-52. doi: 10.1007/s10695-010-9445-0. Epub 2010 Nov 2.
Calcium ions play an important role in the activation of fish sperm movement. The mechanism of their binding in semen is still unknown. The goal of this study was the development of a method for identifying calcium-binding proteins in fish seminal plasma. Two methods of calcium-binding proteins detection were tested with the use of Quin2 and Stains-all dyes. The first method was useful for the identification of calcium-binding proteins of fish seminal plasma. It consisted of proteins separation using SDS-PAGE, transfer on PVDF membrane, incubation with CaCl₂, staining with Quin2 and illumination with UV light to reveal calcium-binding protein bands. Using Quin2 allowed the detection of calcium-binding proteins with low and high molecular weight. Electrophoretic species-specific profiles of calcium-binding proteins were identified in the seminal plasma of carp, whitefish, roach, brook trout, brown trout and rainbow trout. Staining of calcium-binding proteins with Quin2 is a quick and safe method, allowing the identification of calcium-binding proteins in fish semen.
钙离子在鱼类精子运动的激活中起着重要作用。它们在精液中的结合机制尚不清楚。本研究的目的是开发一种鉴定鱼类精液中钙结合蛋白的方法。使用 Quin2 和 Stains-all 染料测试了两种钙结合蛋白检测方法。第一种方法可用于鉴定鱼类精液中的钙结合蛋白。它包括使用 SDS-PAGE 分离蛋白质、转印至 PVDF 膜、用 CaCl₂孵育、用 Quin2 染色和用紫外光照射以显示钙结合蛋白带。使用 Quin2 可以检测到具有低和高分子量的钙结合蛋白。在鲤鱼、白鱼、罗非鱼、溪红点鲑、虹鳟和棕鳟的精液中鉴定出了电泳种特异性钙结合蛋白图谱。用 Quin2 对钙结合蛋白进行染色是一种快速且安全的方法，可用于鉴定鱼类精液中的钙结合蛋白。
