Neuronal activity in motor neuron and V2a interneuron cultures

Somavarapu, Rachel
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The consequences of a spinal cord injury can include paralysis, and loss of motor function, sensory function, and proprioception. Furthermore, spinal cord injuries are often associated with potentially fatal complications, such as pneumonia or sepsis. Spinal cord injuries can take a negative toll on a patient’s life by causing discomfort or by hindering them from living a normal life. Current treatments, such as rehabilitative therapy, electrical spinal cord stimulation, and spinal cord surgery are not effective in treating this condition. However, studying the behavior of cultured neurons in-vitro can help determine what combination of cells are most beneficial for spinal cord transplantation or targeting regeneration of endogenous neural populations. The goal of this project is to study two types of neurons that are found in the spinal cord, motor neurons and V2a interneurons, and examine how these cells behave when they are cultured separately versus cocultured together. Selectable cell lines containing puromycin N-acetyl-transferase (PAC) under the control of either the Hb9 or Chx10 gene regulatory element were induced and selected to form neurons over a 6 day period. The neurons were plated in multielectrode array (MEA) plates or 24 well plates. Collected data regarding the neural firing rate, burst frequency, number of spikes observed, network inter-spike interval coefficient of variation, and synchrony were analyzed to determine differences in activity between mono- and co-cultures.