A Model of Glial Scarring Analogous to the Environment of a Traumatically Injured Spinal Cord Using Kainate.
10.5535/arm.2016.40.5.757
- Author:
Jong Yoon YOO
1
;
Chang Ho HWANG
;
Hea Nam HONG
Author Information
1. Department of Rehabilitation Medicine, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea.
- Publication Type:Original Article
- Keywords:
Spinal cord injuries;
In vitro techniques;
Neuroglia;
Kainic acid
- MeSH:
Animals;
Arm;
Astrocytes;
Chondroitin Sulfate Proteoglycans;
Cicatrix*;
Embryonic Structures;
In Vitro Techniques;
Kainic Acid*;
Neurites;
Neuroglia;
Neurons;
Plastics;
Protein Kinases;
Rats;
Spinal Cord Injuries;
Spinal Cord*;
Vimentin
- From:Annals of Rehabilitation Medicine
2016;40(5):757-768
- CountryRepublic of Korea
- Language:English
-
Abstract:
OBJECTIVE: To develop an in vitro model analogous to the environment of traumatic spinal cord injury (SCI), the authors evaluated change of astrogliosis following treatments with kainate and/or scratch, and degree of neurite outgrowth after treatment with a kainate inhibitor. METHODS: Astrocytes were obtained from the rat spinal cord. Then, 99% of the cells were confirmed to be GFAP-positive astrocytes. For chemical injury, the cells were treated with kainate at different concentrations (10, 50 or 100 µM). For mechanical injury, two kinds of uniform scratches were made using a plastic pipette tip by removing strips of cells. For combined injury (S/K), scratch and kainate were provided. Cord neurons from rat embryos were plated onto culture plates immediately after the three kinds of injuries and some cultures were treated with a kainate inhibitor. RESULTS: Astro-gliosis (glial fibrillary acidic protein [GFAP], vimentin, chondroitin sulfate proteoglycan [CSPG], rho-associated protein kinase [ROCK], and ephrin type-A receptor 4 [EphA4]) was most prominent after treatment with 50 µM kainate and extensive scratch injury in terms of single arm (p<0.001) and in the S/K-induced injury model in view of single or combination (p<0.001). Neurite outgrowth in the seeded spinal cord (β-III tubulin) was the least in the S/K-induced injury model (p<0.001) and this inhibition was reversed by the kainate inhibitor (p<0.001). CONCLUSION: The current in vitro model combining scratch and kainate induced glial scarring and inhibitory molecules and restricted neurite outgrowth very strongly than either the mechanically or chemically-induced injury model; hence, it may be a useful tool for research on SCI.
