| [Objective]:This study aimed to establish the proteomic profile in the spinal cord and its associated precentral gyrus as well as gastrocnemius of rats by using(two-dimensional electrophoresis,2-DE)together with mass spectrometry(MS),then evaluating the relationship between the changes of proteomic expressions and functional restoration as well as spinal cord plasticity following spinal cord transection in the rats.[Materials and Methods]:25 adult SD rats were divided into 5 groups randomly:2-DE condition experiment group,control group,spinal cord transected 3d,14d and 21d groups.Rats in the control group were opened the vertebral canal only,left the spinal cord untouched,while rats in the operated groups were performed spinal cord transected between T10-T11 vertebral levels.Locomotor functions of hindlimbs were assessed by BBB rating scale.Sensory functions were detected by cortical somatosensory evoked potential(CSEP).For 2-DE,total proteins of precentral gyrus,spinal cords and gastrocnemius were extracted,and a Bradford procedure was used to determine the protein concentration in these samples.The proteins from each group were separated by 2-DE and analyzed using PDQuest software.Lastly,the proteins were considered to be significantly regulated(more than 1.5-fold)then followed by MS identification.[Results]:1.In the 2-DE condition experiment,the match rates between different batches of 2-DE maps and the maps from same batch were all more than 70%;2.In the functional assessment,the BBB score of the sham-operated rats were 21,after spinal cord transection,the rats showed complete paralysis(0).However,spontaneous functional recovery of the rats' hindlimbs was gradually detected from 3 to 28 days post operation(dpo).But no CSEP could be recorded throughout the experimental period.3.a,The number of protein spots in the precentral gyrus from control,3,14 and 28dpo groups was 550,412,351 and 357 respectively,and 30 interesting ones were subjected to MS analysis;b,The number of protein spots in the headend spinal cord from the four groups was 500,326,464 and 434 respectively,and 14 significantly regulated ones were detected by MS analysis;c,The number of protein spots in the caudal end spinal cord from the four groups was 580,394,245 and 193 respectively,and 33 differential expressed markedly ones were detected by MS analysis;d,The number of protein spots in the scar from control,14 and 28dpo groups was 553,456 and 263,respectively.While in the other batch,the umber of protein spots in the 14dpo headend and caudal end spinal cords,28dpo headend and caudal end spinal cords as well as 14and 28dpo scars was 557,429,433,400,477 and 397 respectively.Of these proteins,40 interesting ones were detected by MS analysis;e,The number of protein spots in the gastrocnemius from control,3,14 and 28dpo groups was 442,395,407 and 384 respectively,and 18 significantly regulated ones were detected by MS analysis.[Conclusion]:1.We have established the 2-DE technique of precentral gyrus tissue successfully, and the proteins from the spinal cord and its associated precentral gyrus as well as gastrocnemius of rats were also separated and detected successfully using this technique.Together with the functional assessment of hindlimbs,we further revealed that the possible endogenous molecule mechanism of spinal cord plasticity and the relationship between proteomic changes in each associated position and SCI;2.Using the proteomic techniques of 2-DE together with MS,we further explain the relationship between the changes of protein expression and the restoration of injured spinal cord,and proposed that the differential proteins may involved in the mechanisms of functional recovery and spinal cord plasticity following spinal cord transection.
|
PDF Full Text Request