胚胎發育第18天的海馬迴神經細胞與皮質神經細胞中，其最主要的pyramidal neurons都已生成並且具備向其他腦區延伸突起以建立連結的能力。而兩類群的pyramidal neurons在種類、結構及功能都存有差異，使我們希望能了解此時期兩類群神經細胞其蛋白質體的表現情形有何不同。因此我們運用了二維蛋白質電泳搭配coomassie blue染色法呈色膠圖後，先以ImageMaster 2D Platinum 7.0軟體對二維電泳膠圖進行配對分析後，再透過MALDI-TOF鑑定有差異的配對樣點。
本篇研究首先進行二維蛋白質電泳分析條件之前測，為了克服coomassie blue染色法其吸附蛋白質靈敏度不佳的缺陷，我們選擇了增加蛋白質量至250μg，並搭配非線性酸鹼值3到10 IPG strip與10% SDS-PAGE，以此條件進行二維蛋白質電泳分析；我們使用這些改進後的條件對海馬迴神經細胞與皮質神經細胞的蛋白質組成進行分析，並透過2D軟體分析與MALDI-TOF質譜鑑定，發現在兩群組膠圖上有31顆重複出現的配對樣點，未來可搭配2D-DIGE等技術對這些配對樣點進行定量分析。除此之外，我們亦發現了特定表現於海馬迴神經細胞與皮質神經細胞二維電泳膠圖中的蛋白質點，未來可透過MALDI-TOF對這些蛋白質點進行鑑定分析，以進一步了解此時期海馬迴神經細胞與皮質神經細胞其蛋白質體的表現差異。

Bartlett, W. P. and Banker, G. A. (1984). ” An electron microscopic study of the development of axons and dendrites by hippocampal neurons in culture.” J Neurosci. 4(8):1954-65.

Banker, G. A. and Cowan, W. M. (1977). “Rat hippocampal neurons in dispersed cell culture.” Brain Res. 126(3):397-42.

Beaudoin, G. M. 3rd, Lee, S. H., Singh, D., Yuan, Y., Ng Y. G., Reichardt, L. F., Arikkath, J. (2012). ” Culturing pyramidal neurons from the early postnatal mouse hippocampus and cortex.” Nat Protoc. 7(9):1741-54.

Campbell, D. S. and Holt, C. E.(2001). ” Chemotropic responses of retinal growth cones mediated by rapid local protein synthesis and degradation.” Neuron. 32(6):1013-26.

Chevallet, M., Santoni, V., Poinas, A., Rouquié, D., Fuchs, A., Kieffer, S., Rossignol, M., Lunardi, J., Garin, J., Rabilloud, T. (1998). “ New zwitterionic detergents
improve the analysis of membrane proteins by two-dimensional
electrophoresis.” Electrophoresis. 19(11):1901-9.

Curreem, S. O., Watt, R. M., Lau, S. K., Woo, P. C. (2012). “ Two-dimensional gel electrophoresis in bacterial proteomics.” Protein Cell. (5):346-63.

Fishell, G. and Hanashima, C.(2008). “Pyramidal Neurons Grow Up and Change Their Mind.” Neuron. 57, 333-338.

Görg, A., Weiss, W., Dunn, M. J.,(2004). “Current two-dimensional electrophoresis technology for proteomics.” Proteomics. 4(12):3665-85.

Heukeshoven, J. and Dernick, R. (1985).” Simplified method for silver staining of proteins in polyacrylamide gels and the mechanism of silver staining”
Electrophoresis. 6(3):103-112

Jung. H., Byung, C., Yoon, B. C. and Holt,C. E. (2012). “ Axonal mRNA localization and local protein synthesis in nervous system assembly, maintenance and repair” Nature Reviews Neuroscience. 13, 308-324

Kaech, S. and Banker, G.(2006). ” Culturing hippocampal neurons. ” Nat Protoc. 1(5):2406-15.

Klose, J.(1975). “ Protein mapping by combined isoelectric focusing and electrophoresis of mouse tissues. A novel approach to testing for induced point mutations in mammals.” Humangenetik. 26(3):231-43.

Lasek, R. J., Dabrowski, C. and Nordlander, R. (1973). ”Analysis of axoplasmic RNA from invertebrate giant axons.” Nature New Biol. 244, 162–165.

López-Bendito, G. and Molnár, Z. (2003). ” Thalamocortical development: how are we going to get there?” Nature Reviews Neuroscience 4, 276-289

Ming G. L, Wong S. T, Henley J, Yuan X. B, Song H. J, Spitzer N. C, Poo M. M. (2002). “Adaptation in the chemotactic guidance of nerve growth cones.” Nature. 417(6887):411-8.

Molloy, M. P.(2000). “Two-dimensional electrophoresis of membrane proteins using immobilized pH gradients.” Anal Biochem. 280(1):1-10.

Neuhoff, V., Arold, N., Taube, D., Ehrhardt, W. (1988). ” Improved staining of proteins in polyacrylamide gels including isoelectric focusing gels with clear background at nanogram sensitivity using Coomassie Brilliant Blue G-250 and R-250.” Electrophoresis. 9(6):255-62.

O'Farrell, P. H. (1975). “ High resolution two-dimensional electrophoresis of proteins.” J Biol Chem. 250(10):4007-21.

Poland, J., Rabilloud, T., Sinha, P. (2005). “Silver Staining of 2-D Gels” The Proteomics Protocols Handbook , pp 177-184

Rabilloud, T., Luche S., Santoni V., Chevallet M. (2007). “ Detergents and chaotropes for protein solubilization before two-dimensional electrophoresis.” Methods Mol Biol. 355:111-9.

Raymond J. Lasek, Carla Dabrowski and Ruth Nordlander (1973). ” Analysis of
Axoplasmic RNA from Invertebrate Giant Axons” nature new biology 244,162-165.

Samara, A., Vougas, K., Papadopoulou, A., Anastasiadou, E., Baloyanni, N., Paronis, E., Chrousos, G. P., Tsangaris G. T.,(2011). “Proteomics reveal rat hippocampal lateral asymmetry.” Hippocampus. 21(1):108-19.

Spruston, N. (2008). “ Pyramidal neurons: dendritic structure and synaptic integration.” Nat Rev Neurosci. 9(3):206-21

Sun S., Wong E. W., Li M. W., Lee W. M., Cheng C. Y. (2009). “14-3-3 and its binding partners are regulators of protein-protein interactions during spermatogenesis.” J Endocrinol. 202(3):327-36.

Westbrook, J. A, Yan, J. X, Wait, R., Welson, S. Y., Dunn, M. J. (2001). “Zooming-in on the proteome: very narrow-range immobilised pH gradients reveal more protein species and isoforms.” Electrophoresis. 22(14):2865-71.

Wildgruber, R., Harder, A., Obermaier, C., Boguth, G., Weiss, W., Fey, S. J.,Larsen, P. M., Görg, A.,(2000). “Towards higher resolution: two-dimensional electrophoresis of Saccharomyces cerevisiae proteins using overlapping
narrow immobilized pH gradients.Electrophoresis.21(13):2610-6.

Wilkins, M. R., Pasquali, C., Appel, R. D., Ou K., Golaz, O., Sanchez, J. C., Yan, J. X., Gooley, A. A., Hughes, G., Humphery-Smith I., Williams, K. L., Hochstrasser, D. F. (1996). “From proteins to proteomes: large scale protein identification by two-dimensional electrophoresis and amino acid analysis. Biotechnology (N Y). 14(1):61-5.

鄭佳和. (2009). “結合微接觸壓印與模板侷限收基分析海馬迴神經軸突與生長錐
蛋白質” 清華大學 / 生命科學院 / 分子醫學研究所

吳若羚. (2011). ”結合 Lab-on-a-chip與二維蛋白質電泳技術進行海馬迴神經細胞及神經軸突蛋白質組成分析” 清華大學 / 生命科學院 / 分子醫學研究所