Spastin Interact With PDIA6 To Regulate Neurite Outgrowth In Hippocampal Neuron

Objective The regeneration of neurons after spinal cord injury is one of the key points of injury repair.Our laboratory has verified that Spastin plays an important role in promoting the growth of hippocampal neurons.As protein folding enzymes and isomerases,the PDI family also plays an important role in many nervous systems disease.However,whether Spastin and PDI family members can interact and affect the growth of neuronal processes and regulate spinal cord injury repair remains to be explored.The main purpose of this study is whether PDIA6,a member of the PDI family,is involved in the process of spinal cord injury and its repair,and to explore whether Spastin and PDIA6 can interact with each other,and how their interaction affects the growth of hippocampal neurons.Exploring the interaction between Spastin and PDIA6 to mediate the growth of hippocampal neuronal processes will not only help us further understand the mechanism of Spastin’s role in neuronal growth,but also give us a deeper understanding of its role in neurological diseases such as spinal cord injury.It provides a theoretical basis for further research on the repair of spinal cord injury.Materials and methodsConstruct a rat spinal cord injury model,and discover the key proteins in spinal cord injury through proteomic analysis of spinal cord injury samples,tissue section immunofluorescence experiments,and qPCR experiments.Sapstin interacting protein mass spectrometry was used to analyze potential interacting proteins.To make it clear that Spastin can interact with PDIA6 through the construction of Spastin and PDIA6 prokaryotic and eukaryotic expression plasmids,GST Pull-Down experiment and CO-IP experiment.Finally,the relevant eukaryotic plasmids were overexpressed in hippocampal neurons to clarify the effects of Spastin and PDIA6 on the growth of hippocampal neurons.Results(1)Proteomics analysis of spinal cord injury samples,tissue section immunofluorescence experiment,qPCR experiment confirmed that PDIA6 is positively correlated with the severity of spinal cord injury.(2)Through GST-Sapstin interaction protein mass spectrometry analysis,it is clear that PDIA6 is a potential interaction protein of Spastin.(3)It is clear that Spastin can interact with PDIA6 through GST Pull-Down experiment and CO-IP experiment.(4)Co-expression of Spastin and PDIA6 plasmids in primary hippocampal neurons can further promote the growth of hippocampal neurons.Conclusions(1)The expression level of PDIA6 is up-regulated in rat spinal cord injury samples,and the more severe the spinal cord injury,the higher the expression level.(2)The interaction between Spastin and PDIA6 can jointly promote the growth of hippocampal neurons.