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Molecular basis of neuronal adaptation with emphasis on the following adaptive processes; tetanic potentiation, glucocorticoid-induced, age-induced as well as opiate-induced adaptation.
RESEARCH TECHNIQUES
Molecular biology; single-cell genetics; cDNA cloning; in situ hybridization; in situ transcription; mRNA amplification; expression profiling
RESEARCH SUMMARY
The research efforts of my laboratory are directed towards understanding the molecular basis of neuronal functioning. Our experimental approach is reductionist in nature and involves analysis of gene expression in individual cells dispersed in culture, in the live slice preparation or from fixed pathological tissue specimens. We have developed various procedures that have enabled the analysis of cellular functioning using single cells as the experimental model. These procedures include those that permit an analysis of the mRNA complement, the protein complement and an assessment of mRNA movement and translation within single cells. This level of analysis is important since an individual cells biochemical compostion may be diluted by that of surrounding cells. We are currently generating molecular and bioprocess fingerprints of various cell types and disease states. When this is complete, we hope that it will be possible to alter the cellular response to various challenges by altering the levels of these biological processes in a predictable manner. As part of these studies, we are examining the role of subcellular localization of mRNAs in regulating cellular function. We have shown that multiple mRNAs are localized in neuronal dendrites and have provided a formal proof of local mRNA translation in dendrites. Further, we have recently shown that the intracellular sites of localization and translation of these mRNAs can be altered by synaptic stimulation highlighting for the first time that in vivo translation of a mRNA can occur at different rates in distinct regions of a single cell (translation is primarily exponential in dendrites and linear in the cell soma). These insights into the cell biology of neuronal function highlight the complexities that remain to be understood.
Molecular basis of neuronal adaptation with emphasis on the following adaptive processes; tetanic potentiation, glucocorticoid-induced, age-induced as well as opiate-induced adaptation.
RESEARCH TECHNIQUES
Molecular biology; single-cell genetics; cDNA cloning; in situ hybridization; in situ transcription; mRNA amplification; expression profiling
RESEARCH SUMMARY
The research efforts of my laboratory are directed towards understanding the molecular basis of neuronal functioning. Our experimental approach is reductionist in nature and involves analysis of gene expression in individual cells dispersed in culture, in the live slice preparation or from fixed pathological tissue specimens. We have developed various procedures that have enabled the analysis of cellular functioning using single cells as the experimental model. These procedures include those that permit an analysis of the mRNA complement, the protein complement and an assessment of mRNA movement and translation within single cells. This level of analysis is important since an individual cells biochemical compostion may be diluted by that of surrounding cells. We are currently generating molecular and bioprocess fingerprints of various cell types and disease states. When this is complete, we hope that it will be possible to alter the cellular response to various challenges by altering the levels of these biological processes in a predictable manner. As part of these studies, we are examining the role of subcellular localization of mRNAs in regulating cellular function. We have shown that multiple mRNAs are localized in neuronal dendrites and have provided a formal proof of local mRNA translation in dendrites. Further, we have recently shown that the intracellular sites of localization and translation of these mRNAs can be altered by synaptic stimulation highlighting for the first time that in vivo translation of a mRNA can occur at different rates in distinct regions of a single cell (translation is primarily exponential in dendrites and linear in the cell soma). These insights into the cell biology of neuronal function highlight the complexities that remain to be understood.
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