Observation Of The Neurites Growth After Activating Rho Kinase In Cultured Hippocampal Neurons Of Rats With Atomic Force Microscopy

Objective:To invesitgate the process of neurites collapsion by activating the Rho kinase pathway,at the same time observe the changes of the cell membrane structure to enrich the morphology data in neurites formation.Materials and methods:Cultured rat hippocampal neurons were treated with the accelerator of Rho kinase-LPA.The process of neurites extension and tendency of cell membrane changes were analyzed by atomic force microscopy.Results:Neurobasal with B27 has shown excellent long-term viability and nearly pure of postnatal rat hippocampal neurons.When culturing during 6-11 days,hippocampal neurons can be adapted for some other experiments like construction and function.With the development of neurons,the neurites were growing continously and branches getting more and more,the length,diameter and number of the neurons were getting the growth peak when they were cultured 48 hours,and getting declined later.Distinct neurites,enriched cell membrance structures incl-uding membrance proteins and hole-like structures were observed with atomic force microscopy.At the early stage of the neurons development,large quantity of screes-like membrance proteins were deeped in the membrance make the surface rough and uneven.With the development of neurons,the cell membrance were getting more and more enenness and many different shaped hole-like structures were showing up.Length of neurites in cultured neurons treated with LPA were shortened, and the number,length and diameter were shown obvious changes,but similar tendency of the changes were observed both in LPA trested neurons and control cells with the high mature peak at 48 hours.The changes of cell membrance primarily proformanced at relativly long period of uneven surface with quantity of enriched membrance proteins,and also with hole-like structures at the later stage of development,and the number,shape and size of the LPA treate neurons were all different from the control cells.Conclusions:The development of the neurons is a gradual process,neurites continuously grow and extend and branches get more and more and the membrance structures are gradually getting perfect.Activating Rho kinase could induce neurites collapse,and the results were the decreased number of neurites and shortened length.Activating Rho kinase could also induce the structure of the membranc-e change,and the results were the increase of the transport structure of the substance and membrance protein.