earticle

영어

Plasma membrane contains ion channel, protein and peptide receptor, phospholipid and various organelles that play a role metabolism, ion transport, antigen-antibody reaction, hormone response and physiology of reaction in eukaryotic cells. Especially, viability and motility directly related with plasma membrane proteins in sperm. Therefore, this study was to investigate change of proteins by plasma membrane integrity (PMI) in bull sperm. One hundred fifty ejaculates obtained from twenty three bull were diluted in semen extender. Sperm PMI was measured by SYBR14 and propidium iodide (PI) double staining method and analyzed using flow cytometry. Sperm was classified as more than 60% PMI (High PMI) and less than 40% PMI (Low PMI). Proteins were extracted from sperm of each groups using mammalian protein extraction buffer. The proteins was separated using immobilized pH gradient strip by isoelectric point and electrophoresed using SDS-PAGE. Protein intensity of visualized protein spots were detected using image analyzing program, and protein mass was analyzed. Molecular and biological function of proteins was investigated in universal protein resource based on protein mass and coverage rate. In results, 21 protein spots were repeatedly found in the High and Low PMI groups, and molecular functional analysis demonstrated that the proteins were primarily involved in ATP binding and cytokine activity. Especially, MAP kinase mkk-4, cyclin-dependent kinase 1, and heat shock 70 kDa protein 1-like were upregulated, whereas interferon alph-H, interleukin-4, succinyl-Coa ligase subunit beta, and adenylate kinase isoenzyme 1 were down-regulated in the High PMI. Additionally, biological functions demonstrated that proteins involved in cell proliferation and division were up-regulated in the High PMI group. These results may provide understanding of plasma membrane proteins in bull sperm, and will be used in male reproduction for developing specific bio-markers in the future.