Jacobs Sandy, Van de Plas Babs, Van der Gucht Estel, Clerens Stefan, Cnops Lieselotte, Van den Bergh Gert, Arckens Lutgarde
Laboratory of Neuroplasticity and Neuroproteomics, Katholieke Universiteit Leuven, Leuven, Belgium.
Electrophoresis. 2008 Apr;29(7):1518-24. doi: 10.1002/elps.200700487.
To screen for new region-specific protein markers we compared the proteome maps of the primary visual and somatosensory areas V1 and S1 in mouse brain using 2-D difference gel electrophoresis (2-D DIGE). Twenty-three protein spots showed a statistically significant difference in expression level between V1 and S1, with 52% appearing more abundantly in V1. Twenty-six proteins were mass spectrometrically identified in 22 spots. To assess the validity of this list of potential areal markers generated by 2-D DIGE, the effective area-specific distribution profile of creatine kinase brain subtype (CKB), a protein with a clearly higher expression level in S1, was monitored with in situ hybridization. The mRNA expression profile of CKB displayed a clear area-specific distribution, which allowed demarcation of S1 and its topographical borders with neighboring neocortical areas. This proteomic study demonstrates the innovative application of 2-D DIGE and MS to select new regional markers for neuroscience research.
为筛选新的区域特异性蛋白质标志物，我们使用二维差异凝胶电泳（2-D DIGE）比较了小鼠脑初级视觉区和体感区V1和S1的蛋白质组图谱。23个蛋白质点在V1和S1之间的表达水平存在统计学显著差异，其中52%在V1中表达更为丰富。在22个点中通过质谱鉴定出了26种蛋白质。为评估通过二维DIGE生成的这份潜在区域标志物列表的有效性，我们用原位杂交监测了肌酸激酶脑亚型（CKB）的有效区域特异性分布情况，CKB是一种在S1中表达水平明显更高的蛋白质。CKB的mRNA表达谱显示出明显的区域特异性分布，这使得能够划分出S1及其与相邻新皮质区域的拓扑边界。这项蛋白质组学研究证明了二维DIGE和质谱在为神经科学研究选择新的区域标志物方面的创新性应用。
