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
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.
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
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.
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