Western blot detection of brain phosphoproteins after performing Laser Microdissection and Pressure Catapulting (LMPC).
J Neurosci Methods.. 2011-06-15; 198(2): 204-12
DOI: 10.1016/j.jneumeth.2011.04.001
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he Central Nervous System (CNS) is constituted of complex and specific anatomical regions that cluster together and interact with each other with the ultimate objective of receiving and delivering information. This information is characterized by selective biochemical changes that happen within specific brain sub-regions. Most of these changes involve a dynamic balance between kinase and phosphatase activities. The fine-tuning of this kinase/phosphatase balance is thus critical for neuronal adaptation, transition to long-term responses and higher brain functions including specific behaviors. Data emerging from several biological systems may suggest that disruption of this dynamic cell signaling balance within specific brain sub-regions leads to behavioral impairments. Therefore, accurate and powerful techniques are required to study global changes in protein expression levels and protein activities in specific groups of cells. Laser-based systems for tissue microdissection represent a method of choice enabling more accurate proteomic profiling. The goal of this study was to develop a methodological approach using Laser Microdissection and Pressure Catapulting (LMPC) technology combined with an immunoblotting technique in order to specifically detect the expression of phosphoproteins in particular small brain areas.