Experiments | Methods | Markers/Drugs and Time Coursea | No. of Animals |
---|---|---|---|
Sound-induced JNK-mediated mitochondrial cell death pathway | |||
Nature and time course of cell death | Immunocytochemistry | Calbindin, neurofilament, Hoechst | 32 |
TEM | 6 | ||
Cell death mechanisms | Western blots | p-c-Jun | 28 |
Immunocytochemistry | Bax, fodrine, cytochrome c, cytochrome c-Bax-Hoechst cleaved fodrin-calbindin-Hoechst | 18 | |
Protective effects of d-JNKI-1 against sound trauma | |||
Diffusion and penetration | Confocal imaging | 100 μM FITC-d-JNKI-1 | 23 |
Control | Functional assessment | 100 μM TAT-empty, 100 μM JNK-mut | 6 (3 per drugs) |
Protective effects | Dose-response | 0.01-100 μM d-JNKI-1 | 42 (6 per doses) |
SEM | 100 μM d-JNKI-1 | 6 (3 sound exposed untreated + 3 sound-exposed treated) | |
Therapeutic window | Time-response (minipump) | 30 min before and 30 min, 1 h, 4 h, 6 h, 12 h, 24 h after sound exposure. | 42 (6 per time point) |
Time-response (HA gel) | 30 min before and 30 min, 4 h, 12 h after sound exposure | 24 (6 per time point) | |
Western blots | 10 μM d-JNKI-1, 100 μM d-JNKI-1, 100 μM JNK-mut | 16 (4 per drugs and per time points) | |
| Immunocytochemistry | 100 μM of d-JNKI-1 | 6 |
TEM, transmission electron microscopy.
↵ a Markers corresponds to the sound-induced protocol, and Drugs and Time Course corresponds to the protective effects protocol.