2P-24 PDF
Development of a method for analyzing localized lipid metabolism in neurite
Neurite has distinct metabolic compartment from other cellular regions. Recently, some phosphatidylcholines (PCs) were suggested to have characteristic distribution or function in neurites. However, their intracellular local metabolism or specific function is hard to be analyzed because there was technical limitations to control or detect lipid molecules in a single analytical method. Here, we combined MALDI imaging mass spectrometry with a microfluidic device that enables isolation of neurite to develop a new method for analyzing localized lipid metabolism in neurite. Hippocampal neurons corrected from rats at embryonic day 18 were cultured in a microfluidic device on a PLL-coated ITO glass slide. To analyze localized metabolism of exogenous palmitic acid, the neurons in a target side were treated with palmitic acid-d3. Several ions detected directly from neurites were identified as [PC+H]+ ion. When palmitic acid-d3 was administrated to cell body side, PC(32:0-d6) was detected strongly in cell body and weakly in neurites. In contrast, when palmitic acid-d3 was administrated to the distal side, PC(32:0-d6) was strongly detected at tips of the neurites in distal side and not detected in the cell body side. Our method provided direct evidence regarding the distinct metabolic compartment in neurite.