Post-synaptic neuronal responsiveness results from the modulation of neuronal dendritic function. It hasbecome clear over the last several years that various mRNAs are targeted to dendrites where they can belocally translated in response to presynaptic stimulation. The targeting of RNAs into dendrites occursthrough the direct association of RNA binding proteins (RBP) with the RNA being transported. Generally,RBPs not only function as mediators of subcellular localization but also act as chaperones to control themetabolism of RNAs, including transcription of genes, the maturation of RNAs, the nucleo-cytoplasmictransport of RNAs, the translation of RNAs and the degradation of RNAs. The study of populations of RNAsand their regulatory RBPs has been called Ribonomics. In this proposal we propose to assess theRibonomics of dendritic functioning. To do this we will assess three Specific Aims that directly derived fromour preliminary data. In particular we will 1) identify the RBPs that bind to three RNAs that are targeted toand differentially translated in neuronal dendrites, those that encode the GluR2, GluR4 and the NMDA-R1receptor subunit proteins. The identification of the RBPs will be performed using a novel technology that wehave called the PAIR technology. After the RBPs are identified, antibodies directed against these RBPs willbe used in the antibody positioned RNA amplification (APRA) methodology to characterize the family ofother RNAs that bind to the specific RBPs. The combination of the PAIR and APRA methodologies willpermit us to assess changes in these RNAs, their associated RBPs and the RBP co-regulated RNAs duringthe aging process. Secondly, we have recently shown that dendrites have the capacity to modify RNAsthough extra-nuclear RNA splicing. We will continue to investigate this phenomena by identifyingendogenous RNAs that can be locally spliced in dendrites. And finally, using a variety of biochemical andmolecular approaches (including a novel subcellular transfection methodology) a dendrically translated RNAthat gives rise to neuronal cell death. We will assess both fundamental questions of cell death related tothis molecule as well as determine whether there is a potential role for this cell death paradigm inParkinson's Disease and physiological aging. At the conclusion of these studies important roles of RBPsand their cargoes in modulating dendritic function will have been elucidated.
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