Epothilone D Improves Excitatory Synaptic Structure In Cortical Neurons Of BTBR Mouse Model Of Autism In Vitro

Objective:In this study,we evaluate the effect of microtubule stabilizer Epothilone D on excitatory synaptic structure in cortical neurons of autism spectrum disorder(ASD)model of BTBR mouse in vitro,and explore the underlying mechanism.In order to further validate the role of microtubule stability in the pathogenesis of ASD,and provide an experimental and theoretical basis for the further study of Epothilone D on ASD in vivo.Methods:1.The primary cortical neurons of BTBR mouse were isolated and cultured,and the purity of cultured neurons was detected by immunofluorescence technology.2.Cortical neurons of relatively mature were randomly divided into an experimental group and a vehicle control group.In the experimental group,neuronal cultures were treated with 10 n M Epothilone D for 24 h,and the control group given the same volume of solvent DMSO(0.1% v/v DMSO in culture media).After cold stimulation at 4℃ for 90 min,the morphological changes of microtubules were observed by microtubule immunofluorescence staining.3.Western blotting was used to detect the expression of Acetyl-Tubulin,α-Tubulin, MAP2,STOP,VGLUT1,PSD95,Glu N2 B and m Glu R5 proteins.4.Dil staining solution was used to label dendritic spines,and the effect of Epothilone D on the density of dendritic spines was investigated.5.Western blotting and dendritic spine data were analyzed with unpaired t-test (two-tailed)to determine the significant differences between the two groups.For all findings,the statistically significant P values were shown as *P<0.05,**P< 0.01.Results:1.Compared with the solvent control group,10 n M Epothilone D increased the cold stability of microtubules in cortical neurons of BTBR mouse.2.Compared with the solvent control group,10 n M Epothilone D increased the level of the expressions of excitatory synapse-related and glutamate receptor-related proteins in cortical neurons of BTBR mouse.3.Compared with the solvent control group,10 n M Epothilone D increased the density of dendritic spines in cortical neurons of BTBR mouse.Conclusion:1.The microtubule stabilizer Epothilone D improves the excitatory synaptic structure of cortical neurons in BTBR mouse in vitro,which may be related to the increase of microtubule stability.2.The data further provide an experimental and theoretical basis for the study of Epothilone D on the improvement of excitatory synaptic structure and behavior of ASD in vivo.