Introduction
Materials and methods
Experimental animals
Electrode implantation and seizure induction
Rat tissue preparation for RNA isolation and western blot analysis
Cell cultures
Treatment of cell cultures
Preparation of cellular extracts
Western blot analysis
RNA isolation and real-time quantitative PCR analysis (qPCR)
Human material
A II(n = 6) | A III(n = 12) | GBM(n = 55) | Control cortex/autopsy (n = 8) | Control cortex/surgical (n = 5) | |
---|---|---|---|---|---|
Gender (m/f) | 4/2 | 7/5 | 33/22 | 5/3 | 3/2 |
Age (years)a
| 34 (22–43) | 44.7 (33–51) | 56.5 (26–76) | 50 (30–72) | 54 (43–75) |
Location
| - | ||||
Frontal | 5 | 3 | 18 | 4 | 2 |
Temporal | 1 | 3 | 15 | 1 | - |
Parietal | - | - | 4 | 5 | 2 |
Occipital | - | 1 | 3 | 1 | - |
Thalamus | - | - | 1 | - | - |
Parietooccipital | - | 2 | 3 | - | - |
Temporoccipital | - | 2 | 1 | - | 1 |
Temporoparietal | - | - | 1 | - | - |
Frontotemporal | - | 1 | 3 | - | - |
Frontoparietal | - | - | 1 | - | - |
Epilepsy
| 6 | 7 | 29 | - | - |
Duration epilepsy (months)
a
| 6 (4–11) | 4.8 (1–9) | 6.5 (1–12) | - | - |
Tissue preparation for immunocytochemistry
Antibodies
Immunohistochemistry
Evaluation of immunostaining
IR score | A II(n = 6) | A III(n = 12) | GBM (n = 55 ) | Control cortex/autopsy (n = 8) | Control cortex/surgical (n = 5) |
---|---|---|---|---|---|
Kir4.1 | 3.6 ± 0.85* | 7.18 ± 0.41 | 5.5 ± 0.30 | 7.13 ± 0.36 | 7.03 ± 0.26 |
IL-1β | 2.46 ± 0.67* | 3.9 ± 0.9* | 3.75 ± 0.45* | 0 | 0.02 ± 0.02 |
HLA-DR | 5.4 ± 0.77* | 5.7 ± 0.80* | 5.83 ± 0.35* | 0.04 ± 0.02 | 0.03 ± 0.02 |
IR score | With epilepsy (n = 42) | Without epilepsy (n = 31) | With levetiracetam (n = 14 ) | Without levetiracetam (n = 28) |
---|---|---|---|---|
Kir4.1 | 4.9 ± 0.36* | 6.5 ± 0.37 | 6.8 ± 0.62** | 4.5 ± 0.37 |
IL-1β | 5.5 ± 0.49* | 1.3 ± 0.17 | 3.9 ± 1.14** | 6.3 ± 0.46 |
HLA-DR | 5.8 ± 0.31* | 3.6 ± 0.36 | 4.28 ± 0.15 | 5.2 ± 0.40 |