Research Interests:

We use molecular and cell biological methods to assess cell-cell communication in the CNS.

Research Summary:

The research efforts of my laboratory are directed towards understanding the molecular basis of neuronal adaptation. We use both reductionist and expansionist approaches that involve analysis of gene expression in individual cells dispersed in culture or in the live slice preparation as well as coupling of these data with animal behavior. We are characterizing these cells by performing an "expression profiling" of individual neurons and glia in various regions of mouse, rat and human brain. Among the diseases that are studied in the lab are schizophrenia, Alzheimer's disease, Parkinson's disease and Fragile-X syndrome. We have also been attempting to characterize mammalian behavioral genes utilizing the well-characterized operant behavior known as "opiate self-administration". These behavioral studies have been performed in both the rat and mouse systems. Other studies that have a behavioral component are those examining traumatic brain injury in which we are examining the basic molecular underpinnings of neuronal cell death.

Much effort in my lab has been spent characterizing the mRNA complement of a sub-neuronal region called the dendrite. Dendrites are the primary sites at which a pre-synaptic neuron will interact with a post-synaptic neuron. The dendrite can be viewed as "the brain of the neuron". We have shown that there are ~400 mRNAs in dendrites. Further we have provided a molecular proof that these mRNAs can be translated in the dendrite independently of the cell body. The lab's dendrite studies include characterization of 1) dendritic translational regulatory mechanisms, 2) determination of mRNA transport mechanisms and 3) characterization of RNA-binding proteins that facilitate the movement of mRNAs into this subcellular domain.

These studies have been possible because we have developed multiple single cell analysis techniques; including aRNA amplification, in situ transcription, single dendrite transfection assay, antibody positioned RNA amplification and immunodetections amplified by T7 RNA polymerase. We continue to develop methodologies to facilitate our studies including more recently; 1) a novel technology to characterize all of the mRNAs associated with any particular RNA-binding protein, 2) an interacting protein assay that works in vitro and in vivo and 3) a protein profiling methodology that permits the analysis of the protein complement, including post-translational modifications, of even a single cell.

Possible Lab Rotation Projects:

These are numerous and can be designed to address a question of basic science interest or to investigate questions of more immediate clinical significance.

Key References:

Estee Khacharmina, J., Job, C., Crino, P., and Eberwine, J.: Stimulation of Glutamate Receptor Protein Synthesis and Membrane Insertion within Neuronal Dendrites. Proc. Natl. Acad. Sci. 97:11545-11550, 2000.

Eberwine, J., Miyashiro, K., Estee Kacharmina, J. and Job, C.: Local Translation of Classes of mRNAs that are Targetted to Neuronal Dendrites. Proc. Natl. Acad. Sci. 98:7080-7085, 2001

Zhang, H., Kacharmina, J., Miyashiro, K., Greene, M. and Eberwine, J.: Protein Quantification from Complex Protein Mixtures Using a Novel Proteomics Methodology with Single Cell Resolution. Proc. Natl. Acad. Sci. 98:5497-5502, 2001.

Job, C. and Eberwine, J.: Identification of Sites for Exponential Translation in Living Dendrites. Proc. Natl. Acad. Sci., 10.1073/pnas231485698 (early addtion), 2001.

Job, C. and Eberwine, J.: Localization and Translation of mRNA in Dendrites and Axons. Nature Reviews Neuroscience, 2:889-898, 2001.

Miyashiro, K., Beckel-Mitchener, A., Purk, T. P., Belt, B., Kelly, A., Becker, K., Barret, T., Weiler, I.J., Greenough, W. and Eberwine, J.: Deficits in Cellular Functioning Associated with the Loss of FMRP in the CNS of Knockout Mice as Revealed by Antibody Positioned RNA Amplification. Neuron. 37 :417-431, 2003.

Lab Personnel:

David Hinkle, M.D., Ph.D.
Christine Andrews, Ph.D.
Arezou Serabi, Ph.D.
Jason Glanzer, Ph.D.
Kevin Miyashiro
Jennifer Zielinski
Tiina Pajunen
Brian Belt
Jason Davis