Generating A Cortical Stab Injury in a Mouse Model to Induce Reactive Astrogliosis

Abstract

Source: Allahyari, R. V. et al. Triggering Reactive Gliosis In Vivo by a Forebrain Stab Injury. J. Vis. Exp. (2015)

This video demonstrates the cortical stab injury procedure in mice, involving skull exposure and brain lesion creation. It induces reactive astrogliosis, a neuroprotective response of astrocytes, to study CNS trauma, inflammation, and repair mechanisms.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Prepping Mouse for Surgery

1. Remove the mouse from the home cage and weigh (g).
2. Place the mouse into an isoflurane induction chamber and set oxygen to 2 L/min and isoflurane vaporizer to 5 to induce a surgical plane of anesthesia, about 3-5 min. Monitor for slowed breathing and immobilization. Check that the mouse is fully sedated using the toe pinch reflex.
3. When the mouse is fully sedated, place it in a stereotaxic frame, and secure the nose in the....

Results

Figure 1: Schematic of the mouse skull depicting the area of the craniotomy. Blue lines depict the initial markings identifying the boundaries of the area to be drilled. The top and left marks are measured at 1 mm below or lateral to the coronal or sagittal sutu.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Stereotax	Harvard Apparatus	726049
High speed micro drill	Harvard Apparatus	724950
stainless steel scalpel blade, #11	MedVet	JOR581S
5/45 angled forceps	Fine Science Tools	11251-35
Gelfoam sponge 12cmx7mm	Fisher	NC9841478
Antibodies and other reagents	Manufacturer	Catalog Number	Dilution (brightfield) Dilution (fluorescence)
Rb anti-GFAP	DAKO	Z033429-2	1:20k 1:1k
Shp anti-BrdU	Abcam	ab1893	1:2k 1:500
Biotinylated goat anti-rabbit	Vector Laboratories	BA-1000	07:40
Biotinylated rabbit anti-sheep	Vector Laboratories	BA-6000	07:40
Alexafluor 488 goat anti-rabbit	Life Technologies	A-11008	07:40
Alexafluor 568 donkey anti-sheep	Life Technologies	A-21099	17:40
DAPI Nucleic Acid Stain	Life Technologies	D3571	17:40
Cresyl Violet Acetate	Sigma Aldrich	C5042-10G	1%