Objective:Using neuromyelitis optica immunoglobulin G(NMO-IgG)to induced ex vivo mice spinal cord slice model.Methods:Vibratome-cut transverse spinal cord slices from 7-day-old C57BL/6Jmouse pups were cultured on tran...Objective:Using neuromyelitis optica immunoglobulin G(NMO-IgG)to induced ex vivo mice spinal cord slice model.Methods:Vibratome-cut transverse spinal cord slices from 7-day-old C57BL/6Jmouse pups were cultured on transwell porous supports for 7days,then randomly divided into the control group and NMO model group.Slices of the control group were further cultured with human serum complement,while slices from NMO model group were exposed to complement and NMO-IgG.After 24-hour incubation,slices of both groups were measured for aquaporin-4(AQP4),glial fibrillary acidic protein(GFAP),myelin basic protein(MBP)and neurofilament light chain(NFL)by immunofluorescence.Results:Slices exposed to NMO-IgG showed astrocyte swelling,and a significant loss of AQP4and GFAP staining.Ratios of the loss of AQP4and GFAP staining were 77.74%±6.75%and 75.62%±5.76%respectively in the model group,and NMO-like injury score was 3.11±0.60.But there were no obvious losses of AQP4and GFAP staining in the control group,and NMO-like lesion score was 0.00.There were significant differences between the two groups with regards to the above indexes(P<0.01).Ratios of the loss of MBP and NFL staining in the model group were 37.60%±4.88%and46.29%±4.98%respectively,while the corresponding figures in the control group were 9.10%±1.63%and 5.80%±0.81%,and the differences between the two groups were statistically significant(P<0.01).Conclusion:These results suggested that NMO-IgG-induced ex vivo spinal cord slice model possesses typical features of NMO,and this model might be useful for relevant fundamental studies.展开更多
基金supported by the National Natural Science Foundation of China (No.81460194 No.81260188)
文摘Objective:Using neuromyelitis optica immunoglobulin G(NMO-IgG)to induced ex vivo mice spinal cord slice model.Methods:Vibratome-cut transverse spinal cord slices from 7-day-old C57BL/6Jmouse pups were cultured on transwell porous supports for 7days,then randomly divided into the control group and NMO model group.Slices of the control group were further cultured with human serum complement,while slices from NMO model group were exposed to complement and NMO-IgG.After 24-hour incubation,slices of both groups were measured for aquaporin-4(AQP4),glial fibrillary acidic protein(GFAP),myelin basic protein(MBP)and neurofilament light chain(NFL)by immunofluorescence.Results:Slices exposed to NMO-IgG showed astrocyte swelling,and a significant loss of AQP4and GFAP staining.Ratios of the loss of AQP4and GFAP staining were 77.74%±6.75%and 75.62%±5.76%respectively in the model group,and NMO-like injury score was 3.11±0.60.But there were no obvious losses of AQP4and GFAP staining in the control group,and NMO-like lesion score was 0.00.There were significant differences between the two groups with regards to the above indexes(P<0.01).Ratios of the loss of MBP and NFL staining in the model group were 37.60%±4.88%and46.29%±4.98%respectively,while the corresponding figures in the control group were 9.10%±1.63%and 5.80%±0.81%,and the differences between the two groups were statistically significant(P<0.01).Conclusion:These results suggested that NMO-IgG-induced ex vivo spinal cord slice model possesses typical features of NMO,and this model might be useful for relevant fundamental studies.
