Table 1 Overview of in vivo Studies Investigating the Role of Zinc during Cerebral Ischemia

Rat/Global ischemia; 4-VO^a (20 min occlusion)

–

TSQ^b & acid fuschin staining between 2 and 24 h post-ischemia/At 2 h: TSQ-stained somata in CA4, diminished TSQ-signal in mossy fibers; at 18 h: TSQ-stained cells corresponded to acidophilic cells.

(9)

Rat/Focal ischemia; tMCAO^c (60 min occlusion)

ZnPP^d (1–10 µg); topical application

Brain edema (wet/dry weight method) assessed at 24 h post-ischemia/Modest but significant reduction in brain edema with ZnPP treatment.

(24)

Rat/Global ischemia; 4-VO (20 min occlusion)

Pre-tx^e & intra-tx^f hypothermia (29°C) started 1h prior and maintained for ischemia

TSQ staining 2 h to 7 days post-ischemia/lschemia at 37°C: TSQ-stained CA3 neuronal bodies present at 2 to 24 h; ischemia at 29°C TSQ-stained cell bodies absent

(10)

Rat/Focal ischemia; pMCAO^g or tMCAO (2 h occlusion).

ZnPP (50 mg/kg) i.p., pre-tx: 30 min (pMCAO & tMCAO); post-tx^h: 2 or 4 h (tMCAO).

Infarct volume & brain edema assessed 24 h post-ischemia/ZnPP pre-tx in pMCAO: no effect; ZnPP post-tx in tMCAO: no effect; ZnPP pre-tx in tMCAO: dramatic neuroprotective effect reducing infarct volume & brain edema

(26)

Rat/Global ischemia; 2-VOⁱ (15 min occlusion)

Ca-EDTA 300 mM (5 µL; i.c.v.), pre-tx: 30 min.

TSQ & acid fuschin staining 72 h post-ischemia/Zn^l accumulation leads to degeneration; Ca-EDTA reduced Zn accumulation & degeneration of CA1 neurons.

(11)

Rat/Focal ischemia; tMCAO (2 h occlusion)

ZnCI₂^k (10 mg/kg; i.p.), PP^l (48.5 mg/kg; i.p.). ZnPP (50 mg/kg; i.p.), pre-tx: 30 min.

Infarct volume & brain edema assessed 24 h post-ischemia/ZnCI₂ reduced infarct volume but had no effect on brain edema; ZnPP and PP: both significantly reduced infarct volume and brain edema

(27)

Gerbil/Global ischemia; BCCAO^m (3 min occlusion)

ZnCI₂ (20 mg/kg; s.c), pre-tx: 1 h, or 48 and 24 h.

TUNELⁿ and H-E^o at 3 and 4 days, respectively/1 h ZnCI₂ pre-tx: no effect; 48 and 24 h ZnCI₂ pre-tx: modest but significant protection of CA1 region

(28)

Gerbil/Global ischemia; BCCAO (5 min occlusion)

–

Examined ZnT-1 mRNA expression between 12 h to 1 week post ischemia/ZnT-1 mRNA expression was induced in CA1 neurons exhibiting Zn accumulation. Without subsequent ZnT-1 protein expression, these cells started to degenerate at 72 h.

(12)

Rat/Focal ischemia; pMCAO

Neo-Timm stain to show synaptic Zn levels from 7 min to 7 days post-ischemia/Decrease in Zn staining at 7 min and continued throughout 7 days; at 1 h, Zn-positive cell bodies seen

(31)

Rat/Global ischemia; 2-VO (15 min occlusion)

Ca-EDTA 300 mM (3 µL; i.c.v.); pre-tx: 30 min.

TFL-Zn^p and TUNEL staining 24, 48, and 72 h post-ischemia/Ca-EDTA markedly reduced Zn accumulation and degeneration of CA1 neurons at all time points.

(14)

Rat/Global ischemia; 2-VO (12 min occlusion)

Sodium Pyruvate (500 mg/kg; i.p.); pre-tx: 30 min; post-tx: 0⁸, 0.5, 1, 2, 3 h.

TFL-Zn & TUNEL staining 3, 15, and 30 days post-ischemia/Sodium pyruvate almost completely blocked neuronal injury when given within 1 h of ischemia

(16)

Rat/Focal ischemia; tMCAO (30 min occlusion — mild ischemia) or (60 min occlusion — severe ischemia)

Ca-EDTA 500 mM (5 µL; i.c.v.).; pre-tx: 15 min or post-tx: continuous infusion for 1 week (10–100 nmole/h) 1 µ/h; i.c.v.

TSQ, H-E, acid fuschin, CMl-IHC^r staining 3 h to 14 days post-ischemia/Ca-EDTA pre-tx reduced intracellular Zn accumulation, infarct volume, and degeneration at 3 days but protective effects lost at 14 days or if insult intensity increased (30 min occlusion to 60 min occlusion) and if Ca-EDTA was continuously infused for 7 days.

(17)

Mouse/Global ischemia; BCCAO (20 min occlusion).

Hypothermia (33°C) initiated at occlusion; continued 35 min

TSQ & H-E staining 72 h post-ischemia/Hypothermia markedly reduced Zn accumulation and degeneration of CA1, CA2, and CA4 neurons.

(18)

Mouse/Focal ischemia photothrombosis.

–

Infarct volumes & ZnSe^{AMG s} assessed 30 min to 24 h post-ischemia/Infarct core devoid of Zn by 0.5 h until 24 h; 20% increase in Zn in peri-infarct region up to 6 h; Lesions markedly larger at later (12 and 24 h) than at earlier (0.5–6 h) times.

(33)

Rat/Focal ischemia; Embolic MCAO.

ZnCI₂ (80 µmol/kg; i.p.); Bicuculline (48.5 µmol/kg; i.p.) or the above in combination; pre-tx: 30 min

Infarct volume & brain edema assessed 48 h post-ischemia/ZnCI₂ alone or ZnCI₂ and Bicuculline: dramatically increased infarct volume, increased brain edema and worsened neurological deficits; Bicuculline alone: no effect.

(19)

Gerbil/Focal ischemia; right pMCAO.

–

Micro-dialysis collected 0–3 h post-ischemia; analyzed with GF-AAS^t/Zn levels dropped 75% of baseline and never recovered in ipsilateral hemisphere and only minimally changed in contralateral hemisphere

(35)

Gerbil/Global ischemia BCCAO (5 min occlusion) and Rat/Global ischemia 4-VO (10 min occlusion)

Ca-EDTA 300 mM (5 µL; i.c.v.); pre-tx: 30 min or post-tx: 3, 6, 48, 60, or 72 h.

GluR2 mRNA and protein expression, Caspase-3-activity, TSQ, TUNEL 48 h to 5 days post-ischemia/Ca-EDTA pre-tx: reduced Zn levels, GluR2 down-regulation, and early stages of apoptosis in CA1 cells; Ca-EDTA Post-tx between 48 and 60 h: rescued CA1 cells by blocking the later stages of apoptosis (DNA fragmentation).

(22)

Rabbit/Global ischemia; inflatable neck tourniquet & systemic hypoperfusion (30 min occlusion)

–

Micro-dialysis collected 4 to 6 h post-ischemia; analyzed with pZn meter/Ischemia induced immediate rise in extracellular Zn levels from baseline (19 nM) in hippocampus. Reperfusion induced greater rise in extracellular Zn levels (∼100 nM). Glutamate release was earlier and shorter in duration than Zn release

(38)

Rat/Focal ischemia tMCAO (60 min occlusion)

Ca-EDTA 100 mM (5 µL; i.c.v.); pre-tx: 30 min

Infarct volume assessed 3,6, & 24 h post-ischemia/Ca-EDTA pre-tx: deleterious effect on early infarct development: larger infarct volumes at 3 and 6 h but similar to control at 24 h.

(30)

Rat/Global ischemia 4-VO (30 min occlusion)

–

TSQ staining 0 to 24 h post-ischemia/Zn accumulation in CA1 cells at 24 h but not before. Micro-dialysis collected 0 to 3h post-ischemia; analyzed with flameless AAS^U. Extracellular Zn levels in CA1 area increased to ∼600 nM within 15 min of occlusion, decreased during reperfusion, and returned to basal levels (∼300 nM).

(36)

Rat/Focal ischemia tMCAO (60 min occlusion)

–

Micro-dialysis collected 0 to 3 h post-ischemia; analyzed with flameless AAS/Extracellular Zn levels increased to ∼300 nM within 15 min of occlusion, decreased during reperfusion, and returned to basal levels (∼150 nM)

(37)