Ultrasound as a repulsive cue for neuronal development: real-time morphological observations in-vitro

Abstract

BACKGROUND, MOTIVATION AND OBJECTIVE: A core event in neuronal development is the outgrowth of neurite branches from a neuron’s cell body. It is known that, by presenting physical or chemical cues to the extracellular environment, neuronal development can potentially be modulated. In this work, we seek to investigate the use of ultrasound in modifying neuronal development behavior. We hypothesize that ultrasound, when applied at low intensity and pulsed settings, can possibly serve as a physical cue that operates like mechanical stimulation. STATEMENT OF CONTRIBUTION/METHODS: Our study was carried out on a customized platform that allows real-time imaging of neuronal morphology. Pulsed ultrasound (1 MHz frequency, 20 or 100 cycle pulse lengths, 500 or 2500 Hz PRF) was applied through a waveguide at a 45 degree angle with respect to the microscope’s field of view, which is aligned to the center of a 100mm polystyrene cell dish. Three different peak acoustic pressures (measured in-situ) were used: 0.13, 0.40, 0.84 MPa (correspond to SPTA intensities of up to 1.168 W/cm2 for our given pulsing parameters). In this work, N2a mouse neuroblastoma was used as the …