TRPC1 Deletion Causes Dyskinesia And Striatal Proteomic Alterations In Mice

Objective:Transient receptor potential channel 1?TRPC1?is a non-selective channel of Ca²⁺that located on the cell membrane,which can regulates Ca²⁺influx and inhibits neuronal apoptosis induced by endoplasmic reticulum stress.TRPC1 is widely expressed in multiple brain regions and plays an important role in the survival and development of neurons,especially in the tissues that are widely used to study motor dysfunction related to neural,such as basal ganglia,striatum,globus pallidus and substantia nigra.However,the effect of the deletion on the motor ability of the mice and the mechanism was still unknown..The purpose of this study was to explore the effects of TRPC1 deletion on the motion activity and the brain proteomics of mice.Methods:In this work,we selected 11 TRPC1 gene knockout(TRPC1^-/-)mice and the corresponding wild-type?WT?mice as experimental mice.First of all,the open field test,the pole test,the swim test,the rotarod test were employed to evaluate on the motor ability of the TRPC1^-/-mice compared to the WT mice.Afterwards,we used two-dimensional fluorescence difference gel electrophoresis?2D-DIGE?coupled with MALDL-TOF-MS to explore the differentially expressed proteins and the passway of TRPC1^-/-mice compared to that of WT mice.Then,we studied the neuronal loss and apoptosis in striatal by Anti-NeuN immunofluorescent staining,TUNEL immunofluorescent staining and 8-OHdG immunofluorescent staining.Finally,Western-blot analysis was used to verify the expression of some characteristic differential proteins.Results:The results of behavioral test exhibited that the motor ability of TRPC1^-/-mice decreased significantly.The results of immunofluorescence exhibited significantly damage and apoptosis in the nerve cells of striatum of TRPC1^-/-mice.The results of proteomics revealed a total of 51 differentially expressed proteins?26increased and 25 decreased?in stratum of TRPC1^-/-mice compared to WT mice.Bioinformatics analysis exhibited that these dysregulated proteins including:oxidative stress-related proteins,apoptosis-related proteins,endoplasmic reticulum?ER?stress-related proteins,and synaptic proteins,and multiple abnormal expressed proteins were closely related to the development and progression of dysplasia.Among them,oxidative stress-related proteins Protein DJ-1?PARK7?and NDUV2,apoptosis-related proteins Dynamin-1?DYN1?and 14-3-3 protein zeta?14-3-3Z?,endomeric stress related protein GRP78,were verified by Western blot which were consistent with 2D-DIGE.Conclusion:In conclusion,the deletion of TRPC1 gene can lead to dyskinesia and the change of striatum proteome.Among them,the expression change of the molecules such as PARK7,NDUV2,DYN1,14-3-3 zeta and GRP78 may play an important role in studying the effect of the motor behavior and the apoptosis of nerve cells in striatum tissue caused by deletion of TRPC1 gene.