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Table 1 Impact acceleration condition and survival time for each experimental group

From: Targeted disruption of dual leucine zipper kinase and leucine zipper kinase promotes neuronal survival in a model of diffuse traumatic brain injury

Experiments/groupsExperimental history, survival times post-IAProcedures
1.Characterization of initial injury in the optic nerve - generalSham (n = 3), 4 h post-IA (n = 3), 24 h post-IA (n = 3)
IA:40 g × 1 m or 60 g × 0.85 m
IHC for APP (TAI) and IgG (BBB disruption)
2. Characterization of initial injury in the optic nerve - CLARITYCTB injection immediately post-IA, survival 2 d post-IA (n = 3)
IA: 40 g × 1 m -– 60 g × 0.85 m
CLARITY
3. Characterization of traumatic axonopathy - general neuropathology in the optic nerve and tract7 d post-IA (n = 3)
IA:40 g × 1 m or 60 g × 0.85 m
Gallyas silver
IHC for neuroinflammation (IBA1)
4. Characterization of traumatic axonopathy - death of RGCs in retinaSham (n = 6), 2 wk. post-IA (n = 7), 4 wk. post-IA (n = 5)
IA:40 g × 1 m
IHC for γ Synuclein
Cell counts on retinal wholemounts
5. Characterization of traumatic axonopathy - axonal degeneration in optic nerveSham (n = 6), 2 wk. post-IA (n = 7), 4 wk. post-IA (n = 5)
IA:40 g × 1 m
Embedding of optic nerve tissues in epoxy resin, semithin sectioning, toluidine blue staining
6. Induction of DLK-JNK pathway in RGCs after injury24 h (n = 3), 3 d post-IA (n = 3)
IA:40 g × 1 m
IHC for DLK, p-JUN in retinal sections
7. Time course of DLK-JNK induction in RGCs after injurySham (n = 4), 24 h post-IA (n = 10), 3 d post-IA (n = 5), 7 d post-IA (n = 4), 14 d post-IA (n = 3)
IA:40 g × 1 m
IHC for γ Synuclein and p-JUN
Cell counts on retinal wholemounts
8. Time course of activation of distinct member of DLK-JNK pathway in RGCs after injurySham (n = 3), 24 h post-IA (n = 3), 3 d post-IA (n = 3), 7 d post-IA (n = 3)
IA:40 g × 1 m
Standard Western blotting
9. Interventional studies - genetic deletion of Dlk on Dlkfl/fl mice and effects on DLK-JNK pathway activation and RGC survivalAAV2-Cre-GFP into one eye
AAV2-GFP into fellow eye
(n = 4 each, injections 2 wk. pre-IA, euthanasia 3 days post-IA)
IA:60 g × 0.85 m
IHC for p-JUN
Cell counts on retinal wholemounts
AAV2-Cre-GFP into one eye
AAV2-GFP into fellow eye
(n = 7 each, injections 2 wk. pre-IA, euthanasia 4 wk. post-IA)
IA:60 g × 0.85 m
IHC for RBPMS
Cell counts on retinal wholemounts
10. Interventional studies - genetic deletion of Dlk and Lzk on Dlkfl/flLzkfl/fl and effects on DLK-JNK pathway activation and RGC survivalAAV2-Cre-GFP into one eye
AAV2-GFP into fellow eye
(n = 4 each, injections 2 wk. pre-IA, euthanasia 3 day post-IA)
IA:60 g × 0.85 m
IHC for p-JUN
Cell counts on retinal wholemounts
AAV2-Cre-GFP into one eye
AAV2-GFP into fellow eye
(n = 7 each, injections 2 wk. pre-IA, euthanasia 4 wk. post-IA)
IA:60 g × 0.85 m
IHC for RBPMS
Cell counts on retinal wholemounts
11. Interventional studies - pharmacologic kinase inhibition with sunitinib and effects on DLK-JNK pathway activation and RGC survivalDrug (n = 12)
Vehicle (n = 14),
both groups 3 wk. post-IA
IA:40 g × 1 m
IHC for γ Synuclein
Cell counts on retinal wholemounts