In-situ detection of neurotransmitter release from PC12 cells using Surface Enhanced Raman Spectroscopy

Abstract

Rat pheochromocytoma PC12 cells have frequently been used as a dopaminergic neuron model due to their various functions, including the synthesis, storage, and secretion of catecholamines. Furthermore, PC12 cells release a measurable amount of dopamine (DA) in response to some chemicals. PC12 cells are thus considered to be one of the most common invitro models for studying neurotransmitter release. Here, we applied Surface-enhanced Raman Spectroscopy (SERS) to determine with high sensitivity the in-situ short-time effects of cisplatin (cisdiamine- dichloroplatinum), bisphenol-A, and cyclophosphamide on the extracellular DA level released from PC12 cells. In addition, using the SERS technique, changes in the biochemical composition of the PC12 cell lysates were investigated to determine the intracellular DA level. Gold nano-patterned substrates were fabricated based on electrochemical deposition of Au nanorods onto ITO substrates; these substrates were then used as SERS-active surfaces. The Raman spectroscopy results demonstrated that the changes in the Raman spectra depending on the treatment agent were in agreement with the HPLC results on the extracellular DA level. Therefore, the SERS technique can overcome the limitations of other detection techniques, and can be used with cellular nanoarrays to study the effect of a wide range of chemicals.