Cre-LoxP Mediated Induction of Cerebral Cavernous Malformations in a Mouse Model

Samenvatting

Source: Choi, J. P., et al. Induction and micro-CT imaging of cerebral cavernous malformations in mouse model. J. Vis. Exp. (2017)

In this video, a genetically modified mouse pup carrying the Ccm2 gene flanked by loxP sites and Cre recombinase fused to an estrogen receptor is used to induce cerebral cavernous malformations (CCMs). The administration of 4-hydroxytamoxifen activates Cre in endothelial cells, leading to the deletion of Ccm2 and the formation of CCMs in the brain.

Protocol

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

1. Induction of Cerebral Cavernous Malformations or CCMs in Mouse Models

1. Cross Cdh5-CreErt2; Ccm2fl/fl mice with Ccm2fl/fl to generate litters with Cdh5-CreErt2; Ccm2fl/fl (Ccm2iECKO) pups and littermate controls (Ccm2fl/fl). Cdh5-CreErt2 and Ccm2fl/fl animals have been previously described19.
2. Dissolve 4HT in 100% ethanol and store at -80 °C in aliquots of 30 µL 4-hydroxytamoxifen(4HT)( concentration: 10 mg/mL). On the day of use, dilute aliquoted 4HT in corn oil (0.5 mg/mL).
3. Intragastrically inject 50 µL of 4HT (0.5 mg/mL) to neonatal pups at P1 to induce experimental CCM lesions using an insulin syringe.

Materialen

Lijst van materialen gebruikt in dit artikel

Naam	Bedrijf	Catalogusnummer	Opmerkingen
4-hydroxy tamoxifen	Sigma-Aldrich	H6278	To activate Cdh5-CreErt2
Corn oil	Sigma-Aldrich	C8267-500ML	To dilute 4-hydroxy tamoxifen
Insulin syringe