Directing Neuronal Outgrowth and Network Formation of Rat Cortical Neurons by Cyclic Substrate Stretch

Neuronal mechanobiology plays a vital function in brain development and homeostasis with an essential role in neuronal maturation, pathfinding, and differentiation but is also crucial for understanding brain pathology. In this study, we constructed an in vitro system to assess neuronal responses to cyclic strain as a mechanical signal. The selected strain amplitudes mimicked physiological as well as pathological conditions. By subjecting embryonic neuronal cells to cyclic uniaxial strain we could steer the direction of neuronal outgrowth perpendicular to strain direction for all applied amplitudes. A long-term analysis proved maintained growth direction. Moreover, stretched neurons showed an enhanced length, growth, and formation of nascent side branches with most elevated growth rates subsequent to physiological straining. Application of cyclic strain to already formed neurites identified retraction bulbs with destabilized microtubule structures as spontaneous responses. Importantly, neurons were able to adapt to the mechanical signals without induction of cell death and showed a triggered growth behavior when compared to unstretched neurons. The data suggest that cyclic strain plays a critical role in neuronal development.