General Information

Posted 30 October 2005

Transgene: To generate BitetO, a 1.5 kb HindIII fragment corresponding to Xenopus GSK-3 cDNA was excised from a pcDNA3-GSK3 plasmid and subcloned into the pCRII-cloning vector. A 1.5 kb fragment was then excised by NsiI-NotI digestion and subcloned into the PstI-NotI sites of a plasmid containing a bidirectional tetO sequence flanked by CMV minimal promotors with lacZ reporter sequences. Lastly, the 8.0 kb AseI BitetO fragment was microinjected into single-cell CBAxC57BL/6 embryos.

Resulting transgenic mouse lines were generically designated TetO. In the tTA mouse lines, the tTA transgene is under the control of the calcium/calmodulin kinase II promoter. (Mayford, 1996) When the TetO mice are crossed with tTA mice, the resulting double transgenic progeny (designated Tet/GSK-3ß) constitutively express both transgenes.

Mutation: GSK3β

Promoter: tet-responsive promoter for BitetO construct.Calcium/calmodulin kinase IIa promoter for tTA.

Mouse Strain: CBAxC57BL/6

Phenotype

Neuropathological Analysis:

Highest level of transgenic GSK-3β expression is in the hippocampus, then cortex, while little expression could be detected in the striatum. Overexpression of GSK-3β results in neurodegeneration. These tg mice mimic different biochemical and cellular aspects of AD, such as β-catenin destabilization and pretangle-like somatodendritic localization of hyperphosphorylated tau. WB demonstrated increased levels of tau phosphorylation in hippocampus.

Behavior and age of phenotype:

Tet/GSKβ mice were viable and fertile and appeared normal without pharmacological intervention to suppress transgene expression.

Availability

Contact: Jesús Avila
Centro de Biología Molecular "Severo Ochoa," Facultad de Ciencias
Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid, Spain
Phone: +34-91-497-8440
Fax: +34-91-497-4799
Email: Jesús Avila

Patent:
European Patent Application No 01936609.5. Model for neurodegenerative disease. JJ Lucas, F Hernandez, J Avila

References

Primary:

Lucas J J, Hernandez F, Gomez-Ramos P, Moran M A, Hen R, Avila J. Decreased nuclear beta-catenin, tau hyperphosphorylation and neurodegeneration in GSK-3beta conditional transgenic mice. EMBO J. 2001 Jan 15;20(1-2):27-39. Abstract.

Associated:

Mayford M, Bach ME, Huang YY, Wang L, Hawkins RD and Kandel ER. Control of memory formation through regulated expression of a CaMKII transgene. Science, 274, 1678-1683, 1996. Abstract

Engel T, Lucas JJ, Gomez-Ramos P, Moran MA, Avila J, Hernandez F. Co-expression of FTDP-17 tau and GSK-3beta in transgenic mice induce tau polymerization and neurodegeneration. Neurobiol Aging. 2005 Jul 26.