Analysis of cerebellar function by using inducible mGluRl transgenic mice.

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We planed to the role of mGluR1 in generation of neuronal network, synaptic, plasticity, and learning and memory by analysis of the mice, which conditionally express the mGluR1 in the specific brain region. In order to obtain these mice, we generated the mGluR1 transgenic mice in which expression of mGluR1 is regulated by tissue specific promoter and tetracycline or doxycylcine, using transcription factor tTA gene and tTA-responsible enhancer, TRE. We have obtained the following results.1. Generation and analysis of NSE-tTA Tg/lacZ-TRE-mGluR1 Tg/mGluR1 (-/-) : This mice express the mGluR1 in cerebellar Purkinje cells but Tg expression did not rescue the motor discoordination observed in mGluR1 (-/-) mice.2. Generation and analysis of CamKII-tTA Tg/lacZ-TRE-mGluR1a double Tg : These mice die at the ages of 3 to 4 weeks. They can survive and grow normally by application of paste food. The double Tg express mGluR1 mRNA in olfactory bulb, hippocampus CA1 area and striatum, suggesting the ectopic expression of mGluR1 in these regions is responsible for the lethality of the Tg mice.3. Generation of the mice that express tTA in endogenous mGluR1 locus : Since ectopic expression of mGluR1 might lead mice to death, we introduced the tTA gene into mGluR1 locus by gene targeting. We have obtained mGluR1 (+/tTA) mice. Further, we generated 11 independent lines of TRE-mGluR1 Tg.4. Generation of L7 (+/tTA) knock-in mice : We generated the mice that express tTA in cerebellar Purkinje cells. We also made DNA constructs for Tgs expressing TRE-PKCI or TRE-dominant negative MEK, which would inhibit PKC pathway or MAP kinase pathway, respectively.