Matthew B Dalva
Thomas Jefferson University
Project start date: 2007-08-01
Project end date: 2012-06-30
Sponsored Links Excellgen http://Excellgen.com
Grants awarded to Matthew B Dalva
GENETIC INDICATORS FOR DYNAMIC IMAGING OF NEURONAL SIGNALING IN PLASTICITY AND DE
Matthew B Dalva, Assistant Professor
University Of Pennsylvania, 3451 Walnut Street, Philadelphia, Pa 19104
Grant 5R01MH086425-02 from National Institute Of Mental Health
Abstract: During the past decade stunning advances have been made in imaging, molecular biology and biochemistry that enable the visualization of the behavior of single proteins in vivo. Here, I propose to develop and visualize the temporal and spatial dynamics of intracellular signaling within living neurons. Much as early work in calcium imaging redefined our understanding of the importance of calcium influx by defining its spatial and temporal characteristics, I believe visualizing the spatial and temporal dynamics of intracellular signaling will have similar benefits to our understanding of the nervous system. Initially we have developed indicators that enable visualization of one of the key first steps in many intracellular signaling cascades tyrosine phosphorylation. During the past several years, we have developed a system that relies on ratiometric imaging of changes in a genetically encoded fluorescent indicator of phosphorylation. We now propose three specific aims to develop these tools into a system for monitoring signaling during neuronal plasticity and development. We propose to 1) Develop a library of indicators targeted to report activity of specific kinases; 2) Develop indicators that localize to specific cellular compartments; 3) Develop indicators to report activity of multiple signaling molecules simultaneously. Using our indicators, workers will be able to elucidate the dynamics of signals that underlie synaptic plasticity. Thus, our tools will enable novel insights into essential mechanisms that underlie neuronal plasticity. Neuronal plasticity underlies many fundamental functions within the brain, while abnormal neuronal plasticity is associated with disease. Excessive plasticity may underlie diseases like epilepsy and addiction, while defects in plasticity could play important roles in epilepsy, neurodegenerative, and autism spectrum disorders. Our research will have broad impacts across all these levels by developing new tools to visualize dynamic neuronal signaling with subcellular resolution
Keywords: ATP[{..}]protein-tyrosine O-phosphotransferase; Behavior; Biochemical; Biochemistry; Biological; Blood Coagulation Factor IV; Body Tissues; Brain; CNS plasticity; Ca++ element; Calcium; Cell Communication and Signaling; Cell Nucleus; Cell Signaling; Cell membrane; Cells; Characteristics; Chemistry, Biological; Coagulation Factor IV; Color; Coloring Agents; Cytoplasmic Membrane; DNA Molecular Biology; Defect; Degenerative Diseases, Nervous System; Degenerative Neurologic Disorders; Development; Disease; Disorder; Dyes; EC 2.7; EPH- and ELK-Related Tyrosine Kinase; EPH-and ELK-Related Kinase; EPHA8; Encephalon; Encephalons; EphA8 Protein; Ephrin Type-A Receptor 8; Ephrin Type-A Receptor 8 Precursor; Epilepsy; Epileptic Seizures; Epileptics; Event; Factor IV; Family; Genetic; HEK3; Heterogeneity; Image; Imagery; Imaging Procedures; Imaging Techniques; Intracellular Communication and Signaling; Kinases; L-Tyrosine; Laboratories; Libraries; Life; Measurement; Molecular; Molecular Biology; Monitor; NRVS-SYS; Nerve Cells; Nerve Degeneration; Nerve Unit; Nervous System; Nervous System, Brain; Nervous system structure; Neural Cell; Neurocyte; Neurodegenerative Diseases; Neurodegenerative Disorders; Neurologic Body System; Neurologic Degenerative Conditions; Neurologic Diseases, Degenerative; Neurologic Organ System; Neuron Degeneration; Neuronal Plasticity; Neurons; Neurosciences; Nucleus; Organism; PTK; PTK Receptors; Pathway interactions; Phosphorylation; Phosphotransferases; Plasma Membrane; Play; Protein Phosphorylation; Protein Tyrosine Kinase; Protein Tyrosine Kinase EEK; Proteins; RTK; Receptor Protein-Tyrosine Kinases; Reporter; Reporting; Research; Research Design; Resolution; Role; Seizure Disorder; Series; Signal Pathway; Signal Transduction; Signal Transduction Systems; Signaling; Signaling Molecule; Study Type; Synapses; Synaptic; Synaptic plasticity; System; System, LOINC Axis 4; TYR; Technics, Imaging; Testing; Time; Tissues; Transmembrane Receptor Protein Tyrosine Kinase; Transphosphorylases; Tyrosine; Tyrosine Kinase; Tyrosine Kinase Growth Factor Receptor; Tyrosine Kinase Linked Receptors; Tyrosine Kinase Receptors; Tyrosine Phosphorylation; Tyrosine, L-isomer; Tyrosine-Protein Kinase Receptor EEK; Tyrosine-Specific Protein Kinase; Tyrosylprotein Kinase; Visualization; Work; addiction; autism spectrum disorder; base; biological signal transduction; calcium indicator; disease/disorder; epilepsia; epileptiform; epileptogenic; gene product; hydroxyaryl protein kinase; imaging; in vivo; insight; living system; neural degeneration; neural plasticity; neurodegeneration; neurodegenerative illness; neuronal; neuronal degeneration; neuroplasticity; novel; para-Tyrosine; pathway; plasmalemma; public health relevance; ratiometric; single molecule; social role; study design; tool; tyrosyl protein kinase
Relevance: Neuronal plasticity underlies many fundamental functions within the brain, while abnormal neuronal plasticity is associated with disease. Excessive plasticity may underlie diseases like epilepsy and addiction, while defects in plasticity could play important roles in epilepsy, neurodegenerative, and autism spectrum disorders. Our research will have broad impacts across all these levels by developing new tools to visualize dynamic neuronal signaling with subcellular resolution
Project start date: 2009-07-17
Project end date: 2014-04-30
Budget start date: 1-MAY-2010
Budget end date: 30-APR-2011
PFA/PA: RFA-MH-09-030
5R01MH086425-02 (2010): $391950
CRCNS Spontan. Activ. Lateral Interactions & Cort. Maps
Matthew B Dalva
University Of Pennsylvania 3451 Walnut Street Philadelphia, Pa 19104
Grant 5R01MH073357-03 from National Institute Of Mental Health IRG: ZRG1
Abstract: During development, maps of stimuli in the world are established in the cortex. Understanding the role neuronal activity plays in this process has implications for the effects of abnormal sleep, and of toxins and drugs, during pre- and postnatal development in humans. While understanding the underlying cortical circuitry is fundamental to understanding certain neurological pathologies, including mental retardation. Motivated by both experimental and theoretical studies, a prevailing computational model for how maps are formed has been that afferent activity, either spontaneous or environmentally evoked, drives their development. This model assumes that intracortical connections play a subservient role to feedforward ones, that intrinsically generated spontaneous activity does not significantly influence cortical plasticity, and that intraconnections are more or less fixed during development. However, recent data undermine all of these assumptions Modeling studies have shown that maps can arise without patterned feedforward connections. Electrophysiological records of spontaneous activity during initial map formation reveal correlations that may arise from intraconnections patterned by molecular cues. And, photo-stimulation experiments suggest that extensive changes occur in intraconnections during subsequent map development. To test the hypothesis that intrinsic spontaneous activity reinforces pre-existing, rudimentary cortical circuits during early map formation, and reinforces changes in cortical maps during subsequent periods of heightened plasticity, this study will (1) Characterize activity patterns in sleep and wake during initial map formation and subsequent map remodeling in vivo. (2) Map the extent of excitatory and inhibitory intraconnections in vitro. (3) Develop computational models that reproduce the patterns of cortical activity imaged in vivo based on the patterns of lateral connectivity mapped in vitro, and that predict the time-course of map formation and interactions between sleep and wake during map remodeling
Keywords: brain mapping, computer simulation, evoked potential, mathematical model, neural plasticity, photostimulus, synapse, visual feedback, electroencephalography, ferret
Project start date: 2004-08-23
Project end date: 2008-07-31
5R01MH073357-03 (2006): $265589
5R01MH073357-02 (2005): $266101
CELL-CONTACT MEDIATED MECHANISMS ASSEMBLING SYNAPSES
Matthew B Dalva, Assistant Professor
University Of Pennsylvania, 3451 Walnut Street, Philadelphia, Pa 19104
Grant 5R01DA022727-04 from National Institute On Drug Abuse
Abstract: A functional nervous system relies on the formation of precise synaptic connections between specific target neurons. Two key elements of excitatory synapse formation are the formation of mature, mushroom-shaped dendritic spines and the recruitment of synaptic proteins, like NMDARs and PSD-95. A number of adhesion molecules, including EphB/ephrinB, synCAM, SALM2, NGL2, neurexin/neuroligin and cadherins, have each been shown to regulate aspects of synaptogenesis. However, the molecular mechanisms that underlie synapse formation as well as how factors act to coordinate synapse formation in the developing animal remain to be determined. To begin to address these issues, we have focused on the role of the EphB family of receptor tyrosine kinases and their ephrinB ligands because EphB knockout mice show serious deficits in synapse formation, while available knockouts for other synaptic adhesion molecules appear not to have an excitatory synapse formation phenotype. Moreover, because cocaine alters expression of EphB and ephrinB in cortex and striatum, our work on mechanisms of synaptogenesis will provide insights into cortical development and impact our understanding of disease and addiction. We propose that trans-synaptic EphB- ephrinB signaling induces both pre- and post-synaptic organization of multiple components of synapses presynaptic ephrinB, through interactions with dendritic EphB, stimulates postsynaptic development while the reciprocal interaction of postsynaptic EphB induces axonal ephrinB dependent clustering of presynaptic proteins. I propose three specific aims to address the role of EphB and ephrinB in synapse development. Specific aim 1 Determine the signaling mechanisms responsible for EphB-dependent induction of presynaptic specializations. Specific aim 2 Test whether EphB acts to initiate synapse formation. Specific aim 3 Determine the role of EphB proteins in specifying dendritic locations of cortical synaptic contacts in vivo. Our approach of moving from more reduced model systems to the intact animal is innovative and will lead to significant new findings. Moreover, given the role of EphB/ephrinB in synaptic and structural plasticity, and the sensitivity of EphBs and ephrinBs to drugs of abuse, our studies will provide fundamental insights into mechanisms controlling synapse development and with the advent of cell based therapies will advance understanding of human developmental diseases and addiction
Keywords: 8-Azabicyclo(3.2.1)octane-2-carboxylic acid, 3-(benzoyloxy)-8-methyl-, methyl ester, (1R-(exo, exo))-; Abscission; Acute; Address; Adhesion Molecule; Agaricales; Animals; Apical; Assay; Behavior; Bioassay; Biologic Assays; Biological Assay; Biological Models; Biological Neural Networks; Brain; CAM 120/80; Cadherin-1; Cadherins; Cell Adhesion Molecules; Cell Communication and Signaling; Cell Culture Techniques; Cell Function; Cell Process; Cell Signaling; Cell Therapy; Cell physiology; Cells; Cellular Function; Cellular Physiology; Cellular Process; Central Nervous System; Co-culture; Cocaine; Cocultivation; Coculture; Coculture Techniques; Corpus Striatum; Corpus striatum structure; Data; Defect; Degenerative Diseases, Nervous System; Degenerative Neurologic Disorders; Dendrites; Dendritic Spines; Development; Developmentally Regulated EPH-Related Tyrosine Kinase; Disease; Disorder; Drugs; E-Cadherin; ELK-Related Tyrosine Kinase; EPH Tyrosine Kinase 3; Elements; Encephalon; Encephalons; EphB2 Protein; EphB2 Receptor; EphB2-Tyrosine Kinase; Ephrin Receptor EphB2; Ephrin Type-B Receptor 2; Epithelial Calcium-Dependent Adhesion Protein; Epithelial-Cadherin; Escalante syndrome; Excision; Excitatory Synapse; Extirpation; Family; Fragile X; Fragile X Syndrome; Glia; Glial Cells; Human; Human, General; Image; In Vitro; Intracellular Communication and Signaling; Knock-out; Knockout; Knockout Mice; Kolliker`s reticulum; Label; Lead; Ligands; Liver Cell Adhesion Molecules; Location; Mammals, Mice; Man (Taxonomy); Man, Modern; Martin-Bell Syndrome; Martin-Bell-Renpenning syndrome; Mediating; Medication; Membrane; Mice; Mice, Knock-out; Mice, Knockout; Model System; Models, Biologic; Molecular; Morphology; Murine; Mus; Mushrooms; NRVS-SYS; Nerve Cells; Nerve Unit; Nervous System; Nervous System, Brain; Nervous System, CNS; Nervous system structure; Neural Cell; Neuraxis; Neurocyte; Neurodegenerative Diseases; Neurodegenerative Disorders; Neuroglia; Neuroglial Cells; Neurologic Body System; Neurologic Degenerative Conditions; Neurologic Diseases, Degenerative; Neurologic Organ System; Neurons; Non-neuronal cell; Null Mouse; PTK Receptors; Pathology; Pb element; Pharmaceutic Preparations; Pharmaceutical Preparations; Phenotype; Photons; Position; Positioning Attribute; Process; Proteins; Pyramidal neuron; RNA, Small Interfering; RTK; Receptor Protein; Receptor Protein-Tyrosine Kinase HEK5; Receptor Protein-Tyrosine Kinases; Regulation; Removal; Renpenning syndrome 2; Role; Shapes; Signal Transduction; Signal Transduction Systems; Signaling; Site; Slice; Small Interfering RNA; Specific qualifier value; Specified; Spinal Column; Spine; Striate Body; Striatum; Subcellular Process; Surgical Removal; Synapses; Synaptic; System; System, LOINC Axis 4; Testing; Therapy, Cell; Time; Transmembrane Receptor Protein Tyrosine Kinase; Transmission; Tyrosine Kinase Growth Factor Receptor; Tyrosine Kinase Linked Receptors; Tyrosine Kinase Receptors; Tyrosine-Protein Kinase Receptor EPH-3; Uvomorulin; Vertebral column; Work; X-linked mental deficiency-megalotestes syndrome; X-linked mental retardation with fragile X syndrome; X-linked mental retardation-fragile site 1 syndrome; abused drugs; addiction; autism-fragile X (AFRAX) syndrome; autism-fragile X syndrome; backbone; base; biological signal transduction; cell adhesion protein; cell-based therapy; dendrite spine; disease/disorder; drug of abuse; drug/agent; drugs abused; drugs of abuse; experiment; experimental research; experimental study; extracellular; fra(X) syndrome; fra(X)(28) syndrome; fra(X)(q27) syndrome; fra(X)(q27-28) syndrome; fragile X-mental retardation syndrome; fragile Xq syndrome; fragile site mental retardation 1; fragile x [{C0016667}]; fragile x syndromes; gene product; heavy metal Pb; heavy metal lead; hippocampal pyramidal neuron; imaging; in vivo; innovate; innovation; innovative; insight; liver cell adhesion molecule; macro-orchidism-marker X (MOMX) syndrome; macro-orchidism-marker X syndrome; mar(X) syndrome; marker X syndrome; membrane structure; mental retardation-macroorchidism syndrome; mutant; nerve cement; neural network; neurodegenerative illness; neuronal; novel; patch clamp; postsynaptic; presynaptic; receptor; research study; resection; siRNA; social role; striatal; synapse formation; synaptogenesis; tissue fixing; transmission process
Project start date: 2007-08-01
Project end date: 2012-06-30
Budget start date: 1-JUL-2010
Budget end date: 30-JUN-2011
5R01DA022727-04 (2010): $300006
5R01DA022727-03 (2009): $303202
5R01DA022727-02 (2008): $303362
1R01DA022727-01A1 (2007): $309712
Matthew B Dalva
Thomas Jefferson University
Project start date: 2009-07-17
Project end date: 2014-04-30