Molecular basis of neuronal adaptation with emphasis on the following adaptive processes; tetanic potentiation, glucocorticoid-induced, age-induced as well as opiate-induced adaptation.
Molecular biology; single-cell genetics; cDNA cloning; in situ hybridization; in situ transcription; mRNA amplification; expression profiling
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.
Bell TJ, Miyashiro KY, Sul, J-Y, McCullough R, Buckley PT, Jochems J, Meaney DF, Haydon P, Cantor C, Parsons TD, Eberwine, J: Cytoplasmic BK(Ca) channel intron-containing mRNAs contribute to the intrinsic excitability of hippocampal neurons. Proc Natl Acad Sci USA 105: 1901-1906, 2008.
Wu C-K, Zeng, F, Eberwine, J: mRNA transport to and translation in neuronal dendrites [Review] Anal Bioanal Chem 387: 59-62, 2007.
Barrett LE, Sul JY, Takano H, Van Bockstaele EJ, Haydon PG, Eberwine JH: Region-directed phototransfection reveals the functional significance of a dendritically synthesized transcription factor. Nature Methods 3: 455-460, 2006.
Zielinski J, Kilk K, Peritz T, Kannanayakal T, Miyashiro KY, Eiriksdottir E, Jochems J, Langel U, Eberwine J: In vivo identification of ribonucleoprotein-RNA interactions. Proc National Acad Sci USA 103: 1557-1562, 2006.
Zeng F, Peritz T, Kannanayakal TJ, Kilk K, Eiriksdottir E, Langel U, Eberwine J: A protocol for PAIR: PNA-assisted identification of RNA binding proteins in living cells. Nature Protocols 1: 920-7, 2006.
Peritz T, Zeng F, Kannanayakal TJ, Kilk K, Eiriksdottir E, Langel U, Eberwine J: Immunoprecipitation of mRNA-protein complexes. Nature Protocols 1: 577-580, 2006.
Barrett LE, Van Bockstaele EJ, Sul JY, Takano H, Haydon PG, Eberwine JH: Elk-1 associates with the mitochondrial permeability transition pore complex in neurons. Natl Acad Sci USA 103: 5155-5160, 2006.
Glanzer J, Miyashiro KY, Sul J-Y, Barrett L, Belt B, Haydon P, Eberwine J: RNA splicing capability of live neuronal dendrites. Proc Natl Acad Sci USA 102: 16859-16864, 2005.
Marciano PG, Brettschneider J, Manduchi E, Davis JE, Eastman S, Raghupathi R, Saatman KE, Speed TP, Stoeckert CJ Jr, Eberwine JH, McIntosh TK: Neuron-specific mRNA complexity responses during hippocampal apoptosis after traumatic brain injury. J Neurosci 24: 2866-2876, 2004.
Rizzu P, Hinkle DA, Zhukareva V, Bonifati V, Severijnen LA, Martinez D, Ravid R, Kamphorst W, Eberwine JH, Lee VM, Trojanowski JQ, Heutink P: DJ-1 colocalizes with tau inclusions: A link between Parkinsonism and dementia. Ann Neurol 55: 113-118, 2004.
back to top
Last updated: 06/15/2011
The Trustees of the University of Pennsylvania