Study of a pacing neuroblastoma cell line via a novel macroelectrode technique

by 1971- Silva, Ricardo Jose

iii A novel in vitro electrophysiological technique was developed, based on the use of a large summing population electrode (macroelectrode approach). The macroelectrode approach was utilized to study the evolution of the electrical response in the SK-N- BE(2c) human neuroblastoma cell line. Development of a large oscillatory electrical signal consistent with the establishment of a functional pacing network in culture was observed. Neural pacing is a phenomenon of fundamental importance in neurosciences. Pacing is an integrating factor in the developing brain, learning, memory and motor coordination. The nature and origin of this signal was studied by means of the macroelectrode approach as well as patch clamping, liquid chromatography, and immunofluorescence. By comparing the signal obtained with the macroelectrode approach to traditional methods the strength and weaknesses of the technique were explored. It was determined that pacing networks naturally develop in this cell line and are the result of synaptic communication mediated by dopamine as a neurotransmitter and dopamine D2 as a receptor. The macroelectrode technique was shown to provide unique information about the population behavior of cells growing in culture. The electrical properties of the SK-N-BE(2c) cell line as seen with the macroelectrode approach could serve as a cost effective solution for long term, real time study of pacing neural networks in vitro and the basis for the development of cell based biosensors and high throughput drug screening systems.