Studies On The Biophysical Effects Induced By Axotomy For Single Hippocampal Neuron

Single-cell axotomy of hippocampal neurons is a valuable tool for gaining insights into cell injury and axon regeneration.Based on this mode,a discussion of publications dealing with the molecular mechanisms involved in the intrinsic neurite regeneration program have been elucidated.In order to develop a new therapeutics for damaged nerves,much additional studies are required.Numerous studies about neuronal regeneration have focused on biochemical aspects.However,in neuroscience,the physical modifications?such as force?of neuronal injury and injury responses remained to be elucidated.The structural and mechanical properties of living neurons are also essential components that govern many neuronal growth and regeneration processes.Knowledge of the neurite lesion effecting on both mechanical and electrophysiology properties of cultured hippocampal neurons is instrumental to understand the cellular and molecular mechanisms leading to this regenerative growth.This paper presents an overview of the effects of axotomy on hippocampal neurons by employing atomic force microscopy?AFM?-based biomechanical and patch-clamp technique.Firstly,the morphology of cultured hippocampal neurons before and after axotomy were observed.Then,Normal and axotomized hippocampal neurons were measured using advanced AFM force measurement technique and the biomechanical properties of neurons after axotomy were analyzed using a theoretical model.Finally,the voltage-gated Ca²⁺channel currents?VGCCs?were measured by patch-clamp techniques.We observed the cell bodies of neurons after axotomy were smaller than the cell before axotomy,and neurites were degeneration,and the ultrastructure of cell surface changed.The biomechanical results showed that the Young's modulus-E of neurons after axotomy were increased,and the maximum adhesion force of neurons decreased compared with the non-axotomized neurons,respectively.The whole cell patch-clamping assay indicated the VGCCs were diminished significantly.Furthermore,the neurons were cultured after axotomy,new regenerated neurites were visible.Taken together,there was significant recovery of biomechanical and electrophysiology properties after axotomy as time-lapse.Based on the above results,we concluded that axotomy could induce the changes of mechanical and electrophysiological properties of neurons,changing the physiological state of neurons.In addition,the changes of mechanical and electrophysiological properties could feedback the transmission of neuronal signals,which can promote the regeneration of cultural hippocampal neurons.