Visualization of local phosphatidylcholine synthesis within hippocampal neurons using a compartmentalized culture system and imaging mass spectrometry

Highlights: • A compartmentalized neuron culture system was combined with imaging mass spectrometry. • Phosphatidylcholine (PC) species within rat hippocampal neurons were visualized. • Differential synthetic activity of a PC species within neurons was observed. • Neurites synthesized a PC species using local extracellular free fatty acids in situ. Neurons extend neurites with an increased synthesis of phosphatidylcholine (PC) that is not only a membrane component but also a functional regulator with specific fatty acid composition. To analyze the local synthesis of the PC molecular species within neurons, we combined a compartmentalized culture system with matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS). We observed that a newly synthesized PC, which contains exogenously administered palmitic acid-d₃, is accumulated at the cell bodies and the tips of the distal neurites. The local accumulation within distal neurites is formed by distinct metabolic activity from cell bodies, suggesting that the local extracellular composition of free fatty acid can be a key to regulate specific functions of each PC molecular species. We expect our simple method to be a starting point for more sophisticated in vitro analytical methods for unveiling detailed lipid metabolisms within neurons.