| Methodological
approach During the last decade, a number of studies have provided compelling insight into the molecular mechanisms responsible for the induction and stability of synaptic changes and the acquisition and storage of new memories in the mammalian brain despise the complexity and challenging nature of these biological questions. These achievements have been possible thanks to the convergence of very different disciplines and methodologies. I would like to implement this multidisciplinary approach to my research. My laboratory utilizes a bottom-up approach based on the generation of genetically modified mice and their parallel analysis at the biochemical, molecular, electrophysiological and behavioral levels. Specifically, we are using the following methodologies: |
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• Mouse genetics We are currently working with diverse strains of genetically modified mice, including transgenic lines and knockouts. In particular, we use the CaMKII/tTA system of double transgenics developed by Mark Mayford and Eric Kandel. This system allows the expression of genes to be spatially restricted to specific forebrain structures, due to the specificity of the CaMKII promoter, and temporarily dependent on the presence of the drug doxycycline in the mouse diet, which turns off tTA-driven expression. Reversible and restricted transgene expression are critical requirements to reliably assess molecular roles in complex brain functions, such as learning and memory. Additional information about this system and available mouse strains can be found at the Jax® Mice database. |
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• Molecular and cell biology techniques We use conventional techniques to analyze gene expression, such as western-blot, immunohistochemistry, in situ hybridization, quantitative RT-PCR, etc. We are also interested in investigating genome-wide changes in gene expression and transcriptional regulation in vivo. To this end, we are applying recent methodological advances in the field, including last generation high-density oligonucleotide expression arrays from Affymetrix (Mouse Expression 430 2.0 array) to identify transcriptional alterations in mutant mice and chromatin immunoprecipitation (ChIP) assays. |
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• Electrophysiological studies Our analysis of acute hippocampal slices includes measures of excitability, intrinsic membrane properties and synaptic transmission and plasticity. Dr. Mikel Lopez de Armentia, a former member of the team, prepared this interesting introductory video to electrophysiological recordings with the collaboration of Stuart Ingham, INA's graphic designer (sorry, at the moment, soundtrack is only available in Spanish).  |
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• Behavioral studies We have established a battery of behavioral tasks to evaluate basic mouse behavior and, more precisely, to assess explicit forms of memory. This battery includes fear conditioning, spatial learning at the Morris water maze, object recognition, etc... You can see here a video showing how a mouse learns to navigate in the Morris water maze, a common task to evaluate learning and memory in rodents. |
