Experimental Stroke Induction
Objective: To determine whether depleting neuroprogenitor cells (NPCs) affects poststroke functional outcome and recovery of cognitive function in transgenic mice
This is a Experimental Stroke Induction protocol using mouse as the model organism. The procedure involves 5 procedural steps, 2 equipment items, 3 materials. Extracted from a 2013 paper published in Journal of Neuroscience.
Model and subjects
mouse • nestin-δ-HSV-TK-EGFP transgenic mice and wild-type controls • unknown • Not specified • Not specified
Study window
~4 week study window
Core workflow
Osmotic pump implantation and drug administration initiation • Experimental stroke induction • Barnes maze testing for spatial learning and memory
Primary readouts
- Spatial learning and memory performance in Barnes maze
- Poststroke motor function
- NPC ablation in subventricular zone
- Infarct size
Key equipment and reagents
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Protocol Steps
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Osmotic pump implantation and drug administration initiation
Osmotic pumps containing either ganciclovir (200 mg/kg/d) or saline were implanted in nestin-δ-HSV-TK-EGFP transgenic mice and wild-type controls
Note: Administration was continuous for 4 weeks prior to experimental stroke
View evidence from paper
“Ganciclovir (GCV; 200 mg/kg/d) or saline was continuously administered via osmotic pumps in mice for 4 weeks before the induction of experimental stroke”
Experimental stroke induction
Experimental stroke was induced in mice after 4 weeks of ganciclovir or saline administration
Note: Both baseline and stroke-induced type 1 and type 2 NPCs were conditionally ablated by GCV
View evidence from paper
“Both baseline and stroke-induced type 1 and type 2 NPCs were conditionally ablated. GCV eliminated NPCs in a duration-dependent fashion”
Barnes maze testing for spatial learning and memory
Transgenic stroke mice treated with GCV or saline, and wild-type stroke mice treated with GCV were tested in the Barnes maze to assess spatial learning and memory recovery
Note: Transgenic mice given GCV showed impaired spatial learning and memory compared to saline control or wild-type stroke mice given GCV
View evidence from paper
“Transgenic stroke mice given GCV displayed impaired spatial learning and memory in the Barnes maze test compared with saline control or wild-type stroke mice given GCV”
Motor function assessment
Poststroke motor function was evaluated in transgenic mice treated with GCV versus vehicle control
Note: No significant difference in poststroke motor function was observed between groups despite significant NPC ablation
View evidence from paper
“There was no significant difference in poststroke motor function between transgenic mice treated with GCV and those treated with vehicle”
PRV614 injection for retrograde tracing
Polysynaptic viral marker PRV614 was injected into the dentate gyrus (DG) to assess synaptic connectivity between DG and entorhinal cortex (EC)
Note: Retrogradely labeled neurons in the entorhinal cortex were counted to assess synaptic connectivity
View evidence from paper
“nestin-δ-HSV-TK-EGFP mice treated with GCV had fewer retrogradely labeled neurons in the entorhinal cortex (EC) when injected with the polysynaptic viral marker PRV614 in the dentate gyrus (DG)”