William D Snider
University Of North Carolina Chapel Hill
Project start date: 1993-04-07
Project end date: 2017-03-31
Sponsored Links Excellgen http://Excellgen.com
RAF/MEK/ERK REGULATION OF AXON GROWTH AND DIFFERENTIATION
William D Snider, Director, Neuroscience Center
University Of North Carolina Chapel Hill Office Of Sponsored Research Chapel Hill, Nc 27599
Grant 2R01NS031768-14 from National Institute Of Neurological Disorders And Stroke IRG: NDPR
Abstract: The RAF/MEK/ERK signaling cascade is a key mediator of neurotrophin signaling via Trks and is among the most studied signaling cascades in all of biological sciences. In the nervous system, RAF/MEK/ERK signaling is thought to regulate morphological development, differentiation, and neuronal plasticity in response to extracellular signals. Despite its presumed importance, the role of RAF/MEK/ERK signaling in nervous system development and function in vivo is largely unknown due to the fact that the initial round of gene targeted animals has not been informative. We propose a conditional mutagenesis approach to testing the roles of RAF/MEK/ERK signaling on axon growth, differentiation, and regeneration using dorsal root ganglion (DRG) neurons as a model system. Specifically, we propose to test the following five hypotheses /. RAF kinase signaling regulates morphological development but not survival of DRG neurons. II. RAF kinase signaling regulates differentiation of DRG neurons. III. RAF kinase signaling is required for efficient axon regeneration after peripheral axotomy. IV. Conditional elimination of MEKs will yield phenotypes discordant from those produced by conditional elimination of RAFs V Conditional elimination of ERKs will yield phenotypes discordant from those produced by elimination of RAFs and those produced by elimination of MEKs. Defining the biological roles of the RAF/MEK/ERK cascade using conditional gene targeting in mice will provide key insights into the regulation of axon growth, neuronal differentiation and regeneration. The results will also speak to the likely efficacy and potential toxicity of the many drugs being developed to inhibit functions of this cascade for the treatment of neural tumors and chronic pain states.
Project start date: 1993-04-07
Project end date: 2012-03-31
2R01NS031768-14 (2007): $403325
Grants awarded to William D Snider
UNC NEUROSCIENCE CENTER RESEARCH CORES
William D Snider
University Of North Carolina Chapel Hill, Office Of Sponsored Research, Chapel Hill, Nc 27599
Grant 5P30NS045892-08 from National Institute Of Neurological Disorders And Stroke
Abstract: We propose to continue operation of the UNC Neuroscience Center Research Cores funded by the NINDS Institutional Center Core Grant program. These Cores have transformed NINDS-funded research at UNC-Chapel Hill. Five scientific Cores were established in the prior funding period in support of genomics, genetics, and imaging for NINDS-funded investigators. Each of the scientific Cores has been a notable success as manifested by documented intense and broad based usage, and by large numbers of high quality publications based on data obtained using Core equipment and technical support. A number of collaborations among NINDS grantees and between NINDS-grantees and other UNC-Chapel Hill neuroscientists have been inspired by the use of Core services. A well-functioning recharge system is in place for three of the Cores that fully recovers costs from non-NINDS users and provides needed funds for equipment upgrades that cannot be fully funded by this NINDS Center grant. We request continued support of our existing Cores Core 1 Genomics and Bioinformatics (Affymetrix expression profiling, SNP analysis); Core 2 Expression Localization (in situ hybridization); Core 3 BAG Engineering Technology (vector construction in support of mouse genetics); Core 4 ES Cell Technology (ES cell electroporation and characterization) and Core 5 Confocal and Multiphoton Imaging. We propose to improve our existing Cores with new equipment purchases, implementation of the latest research protocols, provision of new services, and in one case expansion into additional space. In addition, we propose a new Core, Core 6 Assay Development for High Throughput Screening that will develop tools for NINDS-funded investigators to interface with the Translational Proteomics Facility at UNC-Chapel Hill and to access High Throughput Screening Centers established by NINDS and other NIH institutes. The UNC Neuroscience Center Research Cores will be managed via an Administrative Core (Core 7). The UNC-Chapel Hill School of Medicine is fully committed to expansion of research in the area of neurological disease. As evidence of this commitment, the UNC-Chapel Hill School of Medicine now commits a substantial package in support of these Cores and in support of new faculty hires in NINDS priority areas
Project start date: 2003-07-01
Project end date: 2013-06-30
Budget start date: 1-JUL-2010
Budget end date: 30-JUN-2011
PFA/PA: PAR-05-070
5P30NS045892-08 (2010): $740000
3P30NS045892-08S1 (2010): $308333
5P30NS045892-07 (2009): $740000
2P30NS045892-06 (2008): $737500
Roles Of Growth Factor Signaling In Neural Regeneration
William D Snider, Director, Neuroscience Center
University Of North Carolina Chapel Hill Office Of Sponsored Research Chapel Hill, Nc 27599
Grant 3R01NS031768-11S1 from National Institute Of Neurological Disorders And Stroke IRG: NDPR
Abstract: This supplemental application requests funds to expand significantly the scope of our original application NS31768 funded April 1,2002. Work sponsored by the grant explores the roles of growth factor signaling mediators in developmental and regenerative axon growth. In Aim 2 of the original grant, we proposed to generate a DRG specific B-Raf null mouse. Increasingly, it is apparent that a more comprehensive analysis of Raf and ERK family members would markedly enhance the impact of our proposal. The Raf/ERK pathway occupies a central position in mediating receptor tyrosine kinase signaling and is expected to be critical in mediating the functions of neurotrophins and other neuronal growth factors throughout the nervous system. The mouse gene targeting studies performed to date have not providing definitive information about requirements for Rafs and ERKs in nervous system development and function, in some cases because of early lethality of null mice, and some cases because of possible "compensatory" effects of closely related family members. Rapid progress in generating floxed alleles using BAC "recombineering" now allows us to target multiple Raf and ERK family members thought to be involved in neurotrophin signaling. We propose a convenient strategy for generating conventional and inducible DRG-specific mutations in B-Raf, C-Raf, ERK2 and ERK5, and for incorporating DRG-specific axonal reporters for convenient visualization of developmental and regenerative sensory axon growth. The supplement requests the funds to support necessary cage costs, PCR genotyping, and colony maintenance. Because of the general importance of neurotrophin signaling to neural development and function, we anticipate that the floxed allele mice we propose to generate will be of interest to other neuroscientists. We will ship any of these lines to investigators who request them at the time of the first publication. Defining the signaling mediators of regenerative axon growth will critically inform strategies to promote regrowth of axons after spinal cord injuries in humans.
Project start date: 1993-04-07
Project end date: 2007-03-31
3R01NS031768-11S1 (2004): $114093
5R01NS031768-13 (2006): $469553
5R01NS031768-12 (2005): $476915
5R01NS031768-11 (2004): $345563
5R01NS031768-10 (2003): $345563
RAF/MEK/ERK REGULATION OF AXON GROWTH AND DIFFERENTIATION
William D Snider
University Of North Carolina Chapel Hill, Office Of Sponsored Research, Chapel Hill, Nc 27599
Grant 5R01NS031768-17 from National Institute Of Neurological Disorders And Stroke
Abstract: The RAF/MEK/ERK signaling cascade is a key mediator of neurotrophin signaling via Trks and is among the most studied signaling cascades in all of biological sciences. In the nervous system, RAF/MEK/ERK signaling is thought to regulate morphological development, differentiation, and neuronal plasticity in response to extracellular signals. Despite its presumed importance, the role of RAF/MEK/ERK signaling in nervous system development and function in vivo is largely unknown due to the fact that the initial round of gene targeted animals has not been informative. We propose a conditional mutagenesis approach to testing the roles of RAF/MEK/ERK signaling on axon growth, differentiation, and regeneration using dorsal root ganglion (DRG) neurons as a model system. Specifically, we propose to test the following five hypotheses /. RAF kinase signaling regulates morphological development but not survival of DRG neurons. II. RAF kinase signaling regulates differentiation of DRG neurons. III. RAF kinase signaling is required for efficient axon regeneration after peripheral axotomy. IV. Conditional elimination of MEKs will yield phenotypes discordant from those produced by conditional elimination of RAFs V Conditional elimination of ERKs will yield phenotypes discordant from those produced by elimination of RAFs and those produced by elimination of MEKs. Defining the biological roles of the RAF/MEK/ERK cascade using conditional gene targeting in mice will provide key insights into the regulation of axon growth, neuronal differentiation and regeneration. The results will also speak to the likely efficacy and potential toxicity of the many drugs being developed to inhibit functions of this cascade for the treatment of neural tumors and chronic pain states
Keywords: Alleles; Allelomorphs; Animals; Axon; Axotomy; Biologic Sciences; Biological; Biological Models; Biological Sciences; CNS plasticity; Cell Communication and Signaling; Cell Signaling; Development; Dorsal Root Ganglia; Drugs; EC 2.7; Embryo; Embryonic; Funding; Ganglia, Spinal; Gene Targeting; Genetics-Mutagenesis; In Vitro; Intracellular Communication and Signaling; Kinases; Life Sciences; MAP-ERK Kinase; MAPK ERK Kinases; MEKs; Mammals, Mice; Mediator; Mediator of Activation; Mediator of activation protein; Medication; Methods; Mice; Model System; Models, Biologic; Molecular Biology, Mutagenesis; Murine; Mus; Mutagenesis; NRVS-SYS; Natural regeneration; Nerve Cells; Nerve Unit; Nervous System; Nervous system structure; Neural Cell; Neural Neoplasm; Neural Tumor; Neurocyte; Neuroepithelial, Perineurial, and Schwann Cell Neoplasm; Neurologic Body System; Neurologic Organ System; Neuronal Differentiation; Neuronal Plasticity; Neurons; Nociception; Pathway interactions; Peripheral; Pharmaceutic Preparations; Pharmaceutical Preparations; Phenotype; Phosphotransferases; Publishing; Regeneration; Regulation; Role; Signal Transduction; Signal Transduction Systems; Signaling; Spinal Ganglia; Targetings, Gene; Testing; Toxic effect; Toxicities; Transphosphorylases; axon growth; axon regeneration; axonal growth; axonal regeneration; biological signal transduction; chronic pain; chronic painful condition; dorsal root ganglion; drug/agent; experience; extracellular; in vivo; insight; mutant; nervous system development; neural plasticity; neuron development; neuronal; neuronal survival; neuroplasticity; neurotrophic factor; neurotrophin; neutrophin; nociceptive; pathway; regenerate; regenerative; response; social role
Project start date: 1993-04-07
Project end date: 2012-03-31
Budget start date: 1-APR-2010
Budget end date: 31-MAR-2011
5R01NS031768-17 (2010): $411131
Sponsored Links Excellgen http://Excellgen.com
5R01NS031768-16 (2009): $407148
3R01NS031768-16S1 (2009): $175414
REGULATION OF DRG DEVELOPMENT BY NEURONAL GROWTH FACTORS
William D Snider, Director, Neuroscience Center
Neurologyuniversity Of North Carolina Chapel Hill
office Of Sponsored Research
chapel Hill, Nc 27599
Grant 5R01NS031768-08 from National Institute Of Neurological Disorders And Stroke IRG: NEUB
Abstract: The purpose of the work outlined is to delineate novel developmental functions of neurotrophins (NTS) and glial-cell line derived neurotrophic factor (GDNF) family members using primary sensory neurons of the dorsal root ganglion (DRG) as a model system. In the prior funding period it was demonstrated that neurotrophin family members are selectively expressed in target fields of different classes of sensory neurons, that nociceptive and proprioceptive DRG neurons express different Trk receptor tyrosine kinases, and that nociceptors and proprioceptors are selectively eliminated in trkA and trkC null mutant mice, respectively. Studies are now proposed which will broaden our understanding of functions of neurotrophins in regulating sensory neuron development and delineate potential interactions with the recently discovered GDNF family of neuronal growth factors. Four questions will be addressed I. Does NT-3 regulate the initial outgrowth and targeting or proprioceptive sensory axons? II. Do levels of NT-3 in developing muscle regulate numbers of spinal cord and brainstem neurons contacted by proprioceptive sensory axons? III. Do levels of NT-3 in spinal cord regulate terminal arborizations of proprioceptive sensory axons in motor pools? IV. Does GDNF act in concert with NT-3 to regulate proprioceptive neuron development? Enhanced understanding of biological actions of neurotrophins and GDNF is of particular importance now that clinical trials have begun assessing these molecules as treatments for peripheral neuropathies and motor neuron disease
Keywords: cell growth regulation, dorsal root, neurogenesis, neurotrophic factor, spinal ganglion axon, developmental neurobiology, growth factor receptor, neuronal transport, protein tyrosine kinase, receptor expression, synaptogenesis in situ hybridization, laboratory rat, transgenic animal
Project start date: 1993-04-07
Project end date: 2002-03-31
5R01NS031768-08 (2000): $264921
7R01NS031768-07 (1999): $257205
UNC Neuroscience Center Research Cores
William D Snider, Director, Neuroscience Center
University Of North Carolina Chapel Hill Office Of Sponsored Research Chapel Hill, Nc 27599
Grant 5P30NS045892-05 from National Institute Of Neurological Disorders And Stroke IRG: NSD
Abstract: The Neuroscience Center at the University of North Carolina-Chapel Hill proposes to establish, house, and staff five Core facilities to support molecular, genetic, and high-resolution imaging approaches for neuroscience research. Four of the proposed Cores will enhance the genetics capabilities of a group of 13 NINDS grantees studying signaling in neural development and neurological disease. They are Core 1. Expression Profiling and Bioinformatics, Core 2. Expression Localization, Core 3. BAC Engineering Technology, and Core 4. Embryonic Stem Cell Technology. These cores will provide integrated services to support molecular genetic and mutagenesis-based approaches. Core 5. Multiphoton and Confocal Imaging, will be jointly established and funded with the Neurodevelopmental Disorders Research Center (NDRC) at UNC. This core will support visualization of migration, axon projection, and dendritic growth in single cells, brain slices and living animals carrying transgenes or mutations generated with the assistance of Cores 3 and 4. The Cores will be housed within space allotted to the Neuroscience Center in a recently completed 8-story Neuroscience Research Building on the School of Medicine campus. The UNC School of Medicine and the North Carolina Biotechnology Center have committed $480,000 toward establishing these Core facilities. The research efforts of 13 NINDS grantees will be supported and extended by the Cores. All of these investigators are members of the UNC Neuroscience Center with joint or primary appointments in basic and clinical departments at the UNC School of Medicine and in the Dept. of Chemistry on the main campus. Eight of the grantees are in adjacent or closely proximate newly constructed laboratory space. Established investigators with expertise in molecular approaches to signaling in the developing or regenerating nervous system will supervise the Cores. In addition, four recently recruited young investigators who are expert in the technologies involved and who are expected to be NINDS grantees in the near future will help direct the Cores and will also be users. A careful operational plan has been developed to govern usage by the group of 13 NINDS grantees with qualifying projects; the four recently recruited young investigators, a group of investigators in the NDRC studying brain development, and the other 15 NINDS grantees on the UNC-Chapel Hill Campus. The quality and relevance of these Core services and their efficient operation will greatly enhance the productivity of NINDS-funded neuroscience research at UNC-Chapel Hill.
Keywords: biomedical facility, neurogenetics, neuroscience
Project start date: 2003-07-01
Project end date: 2008-06-30
5P30NS045892-05 (2007): $708829
5P30NS045892-04 (2006): $730000
5P30NS045892-03 (2005): $730000
5P30NS045892-02 (2004): $730000
1P30NS045892-01 (2003): $654751
RESEARCH TRAINING IN THE NEUROSCIENCES
William D Snider
University Of North Carolina Chapel Hill, Office Of Sponsored Research, Chapel Hill, Nc 27599
Grant 5T32NS007431-12 from National Institute Of Neurological Disorders And Stroke
Abstract: Support is requested to renew and continue a broad, comprehensive, and interdisciplinary predoctoral training program in the neurosciences at the University of North Carolina at Chapel Hill. The training culminates in the award of Ph.D. degree and is administered by the interdepartmental Curriculum in Neurobiology. Training will involve 84 faculty members of the Curriculum (65 Primary Faculty and 19 Associate Faculty), representing research laboratories in 13 departments or programs. Research facilities are well-equipped and funded for a wide variety of cellular, molecular, genetic, biochemical, biophysical, physiological, behavioral, and disease oriented investigations. The Neurobiology Curriculum is closely integrated with the University of North Carolina Neuroscience Center and the program in Behavioral Neuroscience, providing expanded opportunities for training through new research laboratories and the recruitment of new faculty. The formal training program is already in place and constitutes a series of required and elective learning activities. Learning activities include formal coursework, communication skills seminars, laboratory apprenticeships, focused dissertation research under the guidance of faculty mentors, weekly research seminars, clinical correlation experiences, journal clubs, and discussion groups on topics of career development and research integrity. Several annual symposia inspire trainees with presentations by distinguished visiting neuroscientists. An important central goal is to train individuals to utilize methods from a variety of disciplines to probe important problems in neurobiology. The proposed training program will take advantage of several areas of particular strength in neurobiology research at University of North Carolina, including (1) molecular and genetic control of neural development, (2) molecular correlations of specific sensory neuronal function, (3) glial cell biology, (4) structure, function, regulation, and signal transduction pathways of neurotransmitter receptors, (5) functional imaging of nervous system activity in vitro, in vivo, and in situ, and (6) translational research into the biological bases of neurological disease. Trainees take courses in Years -01 and -02, do three research rotations in Year -01, take a written comprehensive qualifying exam at the end of Year -02, defend the dissertation proposal during Year -03, perform intensive research in Years -02 to -06, and defend the dissertation before the end of Year -06 (average 5.7 years). There are now 33 students in the Neurobiology Curriculum and 13 students in Behavioral Neuroscience who are minoring in Neurobiology. An undetermined number of first-year students will enter in fall, 2008. Support is requested for ten predoctoral trainees. Qualified minority candidates are aggressively recruited. RELEVANCE Neuropsychiatric disorders such as Alzheimer´s disease, schizophrenia, chronic pain, and alcohol addiction are a major health burden in the U.S. This Jointly Sponsored NIH Predoctoral Training Program in the Neurosciences supports fundamental, early-stage research training in the neurosciences, leading to the Ph.D. degree. Top students funded by this training program go on to perform cutting-edge research in the neurosciences and make discoveries that lead to new treatments of neuropsychiatric disorders
Keywords: Neurosciences; Research Training
Project start date: 1997-09-30
Project end date: 2014-06-30
Budget start date: 1-JUL-2010
Budget end date: 30-JUN-2011
PFA/PA: PAR-08-101
5T32NS007431-12 (2010): $408720
Sponsored Links Excellgen http://Excellgen.com
2T32NS007431-11A1 (2009): $341178
GSK-3B/APC In Developmental & Regenerative Axon Growth
William D Snider, Director, Neuroscience Center
Neurologyuniversity Of North Carolina Chapel Hill
office Of Sponsored Research
chapel Hill, Nc 27599
Grant 5R01NS050968-05 from National Institute Of Neurological Disorders And Stroke IRG: NDPR
Abstract: Links between neurotrophin signals that regulate neuronal morphology and the neuronal cytoskeleton have remained elusive. In vitro, we have identified a pathway downstream of NGF involving spatially localized PI3K and GSK-3b signaling and binding of the Adenomatous Polyposis Coli protein (APC) to microtubule + ends at the growth cone. We hypothesize that this pathway mediates microtubule assembly during NGF induced axon growth. Both APC and GSK-3b are also strikingly localized to the distal axon tips and growth cones of rapidly growing axons of "precondition lesioned" DRG neurons in vitro. Thus we hypothesize the GSK-3b/APC pathway mediates microtubule assembly in regenerative axon growth as well. In vivo functions of GSK-3b and APC related to nervous system development have not yet been explored because of embryonic lethality in gene targeted mice and because each has a related family member also heavily expressed in the mammalian nervous system that may have a "compensatory" function. We plan to address in vivo roles of these proteins in the current proposal. We will generate inducible, DRG-specific knock-outs for GSK-3a, GSK-3b, APC, APC-L, and an upstream kinase, ILK. Using DRG specific axonal reporter mice, we will determine the roles of the GSK-3b/APC pathway in the development of peripheral and spinal cord DRG projections and in the axon regeneration normally induced by a peripheral nerve crush. If GSK-3b and APC are found to be required for assembly of microtubules during axon regeneration in vivo, and if key upstream regulators can be identified, our studies would provide a new pharmacological approach to enhancing axon regeneration after injur
Keywords: axon, cell morphology, cytoskeletal protein, nerve growth factor, nervous system regeneration, neuronal transport, phosphatidylinositol 3 kinase, serine threonine protein kinase biological signal transduction, growth factor receptor, integrin, nerve /myelin protein, neurobiology, neurogenesis genetically modified animal, laboratory mouse, polymerase chain reaction, site directed mutagenesis, southern blotting
Project start date: 2004-09-15
Project end date: 2010-04-30
5R01NS050968-05 (2008): $320130
5R01NS050968-04 (2007): $320130
5R01NS050968-03 (2006): $329691
5R01NS050968-02 (2005): $337625
GSK-3B/APC In Developmental And Regenerative Axon Growth
William D Snider, Director, Neuroscience Center
University Of North Carolina Chapel Hill Office Of Sponsored Research Chapel Hill, Nc 27599
Grant 1R01NS050968-01 from National Institute Of Neurological Disorders And Stroke IRG: NDPR
Abstract: Links between neurotrophin signals that regulate neuronal morphology and the neuronal cytoskeleton have remained elusive. In vitro, we have identified a pathway downstream of NGF involving spatially localized PI3K and GSK-3b signaling and binding of the Adenomatous Polyposis Coli protein (APC) to microtubule + ends at the growth cone. We hypothesize that this pathway mediates microtubule assembly during NGF induced axon growth. Both APC and GSK-3b are also strikingly localized to the distal axon tips and growth cones of rapidly growing axons of "precondition lesioned" DRG neurons in vitro. Thus we hypothesize the GSK-3b/APC pathway mediates microtubule assembly in regenerative axon growth as well. In vivo functions of GSK-3b and APC related to nervous system development have not yet been explored because of embryonic lethality in gene targeted mice and because each has a related family member also heavily expressed in the mammalian nervous system that may have a "compensatory" function. We plan to address in vivo roles of these proteins in the current proposal. We will generate inducible, DRG-specific knock-outs for GSK-3a, GSK-3b, APC, APC-L, and an upstream kinase, ILK. Using DRG specific axonal reporter mice, we will determine the roles of the GSK-3b/APC pathway in the development of peripheral and spinal cord DRG projections and in the axon regeneration normally induced by a peripheral nerve crush. If GSK-3b and APC are found to be required for assembly of microtubules during axon regeneration in vivo, and if key upstream regulators can be identified, our studies would provide a new pharmacological approach to enhancing axon regeneration after injur
Keywords: axon, cell morphology, cytoskeletal protein, nerve growth factor, nervous system regeneration, neuronal transport, phosphatidylinositol 3 kinase, serine threonine protein kinase, biological signal transduction, growth factor receptor, integrin, nerve /myelin protein, neurobiology, neurogenesis, genetically modified animal, laboratory mouse, polymerase chain reaction, site directed mutagenesis, southern blotting
Project start date: 2004-09-15
Project end date: 2008-04-30
1R01NS050968-01 (2004): $337625
GSK-3 IS A MASTER REGULATOR OF NEURAL PROGENITOR SELF-RENEWAL
William D Snider
University Of North Carolina Chapel Hill, Office Of Sponsored Research, Chapel Hill, Nc 27599
Grant 2R01NS050968-06 from National Institute Of Neurological Disorders And Stroke
Abstract: In the prior funding period, we demonstrated that conditional elimination of the Glycogen Synthase Kinase-3s (GSK-3s), a and ¿, in the embryonic nervous system results in remarkable dysregulation of neural progenitor homeostasis. This result has important translational significance because of the widespread use of a GSK-3 inhibitor, lithium, in clinical practice. We hypothesize that inactivation of GSK-3s renders progenitors incapable of responding to the extracellular signals that normally regulate conversion of radial progenitors to neurons and intermediate neuronal precursors (INPs). We now propose definitive mouse genetic experiments to assess mechanisms of this GSK-3 regulation (Aims I and II). Further, we will use a chemical genetics approach to determine if reinduction of GSK-3 activity in GSK-3 deficient progenitors will result in enhanced neurogenesis (Aim III). Finally we will ask whether GSK-3 signaling regulates neural progenitors in the adult dentate gyrus (Aim IV). This work will reveal the functional potential and mechanisms of GSK-3 regulation of neural progenitors in mammals. The work will also provide information on previously unrecognized potential effects of lithium a drug commonly used in clinical practice, increasingly in children. Finally our results may suggest new ways to expand neural progenitor populations in the setting of neural transplantation. We propose that a specific protein, GSK-3, is a master regulator of neural stem cells. This function of GSK-3 requires investigation because a GSK-3 inhibitor, lithium, is commonly used in clinical practice, increasingly in children. Our results may also suggest new ways to expand neural stem cells in the setting of neural transplantation
Keywords: 0-11 years old; 21+ years old; Adult; Alleles; Allelomorphs; Ammon Horn; Autoregulation; Brain; Breeding; CCI-779; Cell Communication and Signaling; Cell Count; Cell Cycle; Cell Cycle Inhibitor 779; Cell Division Cycle; Cell Growth in Number; Cell Multiplication; Cell Number; Cell Proliferation; Cell Signaling; Cellular Proliferation; Chemicals; Child; Child Youth; Children (0-21); Cornu Ammonis; DNA Recombination; DNA recombination (naturally occurring); Dentate Fascia; Dentate Gyrus; Development; Dorsal; Drugs; EC 2.7; Embryo; Embryonic; Embryonic Nervous System; Encephalon; Encephalons; Erinaceidae; Exhibits; Family member; Fascia Dentata; Funding; GSK-3; Genetic; Genetic Recombination; Genetics-Mutagenesis; Glycogen Synthase Kinase 3; Glycogen Synthase Kinases; Gyrus Dentatus; Hedgehogs; Hippocampus; Hippocampus (Brain); Homeostasis; Human, Adult; Human, Child; Intracellular Communication and Signaling; Investigation; Kinases; Knock-in; Knock-in Mouse; Li+ element; Life; Lithium; Mammalia; Mammals; Mammals, General; Mammals, Mice; Mediating; Mediator; Mediator of Activation; Mediator of activation protein; Medication; Mice; Molecular Biology, Mutagenesis; Mother Cells; Murine; Mus; Mutagenesis; Nerve Cells; Nerve Unit; Nervous; Nervous System, Brain; Neural Cell; Neural Growth; Neural Stem Cell; Neurocyte; Neuronal Differentiation; Neuronal Growth; Neurons; Notch Signaling Pathway; PTK Receptors; Pharmaceutic Preparations; Pharmaceutical Preparations; Phosphotransferases; Physiological Homeostasis; Point Mutation; Population; Position; Positioning Attribute; Progenitor Cells; Proteins; Proteolysis and Signaling Pathway of Notch; RTK; Radial; Rapamune; Rapamycin; Rapamycin Analog; Rapamycin Analog CCI-779; Receptor Protein-Tyrosine Kinases; Recombination; Recombination, Genetic; Regulation; Signal Transduction; Signal Transduction Systems; Signaling; Sirolimus; Stem cells; Structure of dentate gyrus; Telencephalon; Testing; Transmembrane Receptor Protein Tyrosine Kinase; Transphosphorylases; Transplantation; Tyrosine Kinase Growth Factor Receptor; Tyrosine Kinase Linked Receptors; Tyrosine Kinase Receptors; Work; adult human (21+); biological signal transduction; cell imaging; cell type; cellular imaging; chemical genetics; children; clinical practice; dentate gyrus; drug/agent; experiment; experimental research; experimental study; extracellular; gene product; granule cell; gsk-3 Gene Product; hippocampal; in vivo; inhibitor; inhibitor/antagonist; nerve stem cell; nestin; nestin protein; neural; neural progenitor cells; neurogenesis; neuronal; neuronal progenitor; neuronal progenitor cells; notch; notch protein; notch receptors; postnatal; progenitor; public health relevance; relating to nervous system; research study; self-renewal; transplant; youngster
Relevance: Narrative Statement We propose that a specific protein, GSK-3, is a master regulator of neural stem cells. This function of GSK-3 requires investigation because a GSK-3 inhibitor, lithium, is commonly used in clinical practice, increasingly in children. Our results may also suggest new ways to expand neural stem cells in the setting of neural transplantation
Project start date: 2004-09-15
Project end date: 2015-01-31
Budget start date: 1-FEB-2010
Budget end date: 31-JAN-2011
PFA/PA: PA-07-070
2R01NS050968-06 (2010): $397432
TROPIC FUNCTIONS OF NEUROTROPHINS
William D Snider
Washington University 1 Brookings Dr, Campus Box 1054 Saint Louis, Mo 631304899
Grant 5P01NS034448-050004 from National Institute Of Neurological Disorders And Stroke
Abstract: from the ) Dr. Snider proposes to examine the role of NT3 in guiding axons by creating focal sources of this neurotrophin and by disrupting endogenous gradients and focal sources of this same neurotrophin. Specifically, he will focus on peripheral and central projections of Ia afferent sensory neurons, a neuronal population known to be dependent on NT3 for survival and to express the major NT3 receptor trkC. Specific experiments to assess effects of NT3 on axonal growth and branching by creating sources of NT3 will (1) express NT3 under the myogenenin promoter to assess effects on the projection to skeletal muscles (and presumed absence of effects on the central projection); (2) target NT3 to motoneurons and white matter, using the choline-o-acetyltransferase and myelin basic protein promoters, respectively. These sources are predicted to result in aberrant growth to preganglionic autonomic neurons and aberrant branching, respectively; (3) similar targeting of NT4 and BDNF to motoneurons using the ChAT promoter which is predicted to induce projections from other sensory neurons that do not normally innervate ventral spinal cord; (4) Crossing of the myogenin-NT3 mice with NT3 mutants which is predicted to spare Ia efferents but disrupt the postulated NT3 gradient in spinal cord, resulting in loss or aberrancy in the central projection; (5) Ablation of motoneurons with a ChAT-Diphtheria toxin construct which is predicted to reduce or eliminate the Ia projection to the central cord.
Keywords: axon, developmental neurobiology, neuronal guidance, transfection, dorsal root, gene expression, genetic promoter element, genotype, motor neuron, confocal scanning microscopy, immunocytochemistry, in situ hybridization, laboratory mouse, polymerase chain reaction, staining, transgenic animal
GDNF AND SYNAPTIC COMPETITION AND MAINTENANCE
William D Snider, Director, Neuroscience Center
Neurologyuniversity Of North Carolina Chapel Hill
office Of Sponsored Research
chapel Hill, Nc 27599
Grant 5R37NS037873-04 from National Institute Of Neurological Disorders And Stroke IRG: NEUB
Abstract: adapted from ´s ) It has long been hypothesized that synaptic rearrangements in the developing nervous system are mediated by competition between axons for a limited supply of target-derived neurotrophic molecules. However, there has been little direct evidence indicating local maintenance of synapses by retrogradely acting factors. One site where target-derived factors might be playing a pivotal role is at the developing neuromuscular junction where synapses from all but one axon are eliminated in early postnatal life through a competitive mechanism that is little understood. The developing neuromuscular junction is a powerful system in which to study synaptic competition as boutons from competing axons can be visualized and assessed physiologically as they are eliminated over the first two postnatal weeks. Glial-cell line derived neurotrophic factor (GDNF) is a potent survival factor for motor neurons. Our preliminary data show that transgenic overexpression of GDNF in developing muscle leads to dramatic hyperinnervation at motor endplates. This effect is specific for GDNF as overexpression of several neurotrophins also known to promote motorneuron survival does not influence the number of axons converging at the neuromuscular junction. In five specific aims we now propose to explore the mechanism by which GDNF causes hyperinnervation, ask whether long-term excess of GDNF can permanently prevent naturally-occurring synapse elimination, and test the hypothesis that GDNF (or a GDNF family member) regulates synapse elimination during normal development. To accomplish these aims, we have developed new techniques including an intracellular injection method for introducing foreign genes into single muscle fibers in vivo and a method of transplanting muscle from null mice into normal hosts. Determining how target-derived trophic factors influence synaptic competition on a local level will advance our understanding of the stability and plasticity of synaptic circuits. The dramatic effects of GDNF on muscle fiber innervation suggest a role for this molecule in the treatment of denervating conditions such as motor neuron diseases, neuropathies, and trauma
Keywords: innervation, neuromuscular junction, neurotrophic factor, synapse gene expression, growth factor receptor, protein localization confocal scanning microscopy, immunocytochemistry, laboratory mouse, muscle transplantation, transfection, transgenic animal
Project start date: 1998-07-06
Project end date: 2003-06-30
5R37NS037873-04 (2001): $362444
5R37NS037873-03 (2000): $354220
Sponsored Links Excellgen http://Excellgen.com
7R37NS037873-02 (1999): $357219
William D Snider
University Of North Carolina Chapel Hill
Project start date: 2003-07-01
Project end date: 2013-11-30
William D Snider
University Of North Carolina Chapel Hill, Office Of Sponsored Research, Chapel Hill, Nc 27599
Keywords: Accounting; Budgets; Computer Programs; Computer software; Core Facility; Data; Equipment; Faculty; Fees; Funding; Grant; Internet; Maintenance; Maintenances; Monitor; Neurosciences; Operation; Operative Procedures; Operative Surgical Procedures; Pattern; Procedures; Qualifying; Reagent; Research; Software; Structure; Surgical; Surgical Interventions; Surgical Procedure; Universities; Update; WWW; computer program/software; design; designing; meetings; mouse model; surgery; web; world wide web
Budget start date: 1-JUL-2010
Budget end date: 30-JUN-2011
5P30NS045892-08_9006 (2010): $32684
5P30NS045892-07_9006 (2009): $32684
Core--Multiphoton And Confocal Imaging
William D Snider, Director, Neuroscience Center
University Of North Carolina Chapel Hill Office Of Sponsored Research Chapel Hill, Nc 27599
Grant 5P30NS045892-059005 from National Institute Of Neurological Disorders And Stroke IRG: NSD
Keywords: biomedical facility, neuroimaging, confocal scanning microscopy, developmental neurobiology, fluorescence microscopy, fluorescent dye /probe, intravital microscopy, bioimaging /biomedical imaging, laboratory mouse
William D Snider, Director, Neuroscience Center
University Of North Carolina Chapel Hill Office Of Sponsored Research Chapel Hill, Nc 27599
Grant 5P30NS045892-059002 from National Institute Of Neurological Disorders And Stroke IRG: NSD
Keywords: biomedical facility, gene expression, in situ hybridization, neurogenetics, nucleic acid probe
Core--Multiphoton And Confocal Imaging
William D Snider, Director, Neuroscience Center
University Of North Carolina Chapel Hill Office Of Sponsored Research Chapel Hill, Nc 27599
Grant 5P30NS045892-049005 from National Institute Of Neurological Disorders And Stroke IRG: NSD
Keywords: biomedical facility, neuroimaging, confocal scanning microscopy, developmental neurobiology, fluorescence microscopy, fluorescent dye /probe, intravital microscopy, bioimaging /biomedical imaging, laboratory mouse
William D Snider, Director, Neuroscience Center
University Of North Carolina Chapel Hill Office Of Sponsored Research Chapel Hill, Nc 27599
Grant 5P30NS045892-049002 from National Institute Of Neurological Disorders And Stroke IRG: NSD
Keywords: biomedical facility, gene expression, in situ hybridization, neurogenetics, nucleic acid probe
Sponsored Links Excellgen http://Excellgen.com
Core--Multiphoton And Confocal Imaging
William D Snider, Director, Neuroscience Center
University Of North Carolina Chapel Hill Office Of Sponsored Research Chapel Hill, Nc 27599
Grant 5P30NS045892-039005 from National Institute Of Neurological Disorders And Stroke IRG: NSD
Keywords: biomedical facility, neuroimaging, confocal scanning microscopy, developmental neurobiology, fluorescence microscopy, fluorescent dye /probe, intravital microscopy, bioimaging /biomedical imaging, laboratory mouse
GDNF AND SYNAPTIC COMPETITION AND MAINTENANCE
William D Snider, Director, Neuroscience Center
University Of North Carolina Chapel Hill Office Of Sponsored Research Chapel Hill, Nc 27599
Grant 5R37NS037873-07 from National Institute Of Neurological Disorders And Stroke IRG: NSS
Keywords: innervation, neuromuscular junction, neurotrophic factor, synapse, gene expression, growth factor receptor, protein localization, confocal scanning microscopy, genetically modified animal, immunocytochemistry, laboratory mouse, muscle transplantation, transfection
Project start date: 1998-07-06
Project end date: 2006-04-30
5R37NS037873-07 (2004): $420532
William D Snider, Director, Neuroscience Center
University Of North Carolina Chapel Hill Office Of Sponsored Research Chapel Hill, Nc 27599
Grant 5P30NS045892-029002 from National Institute Of Neurological Disorders And Stroke IRG: NSD
Keywords: biomedical facility, gene expression, in situ hybridization, neurogenetics, nucleic acid probe
Core--Multiphoton And Confocal Imaging
William D Snider, Director, Neuroscience Center
University Of North Carolina Chapel Hill Office Of Sponsored Research Chapel Hill, Nc 27599
Grant 5P30NS045892-029005 from National Institute Of Neurological Disorders And Stroke IRG: NSD
Keywords: biomedical facility, neuroimaging, confocal scanning microscopy, developmental neurobiology, fluorescence microscopy, fluorescent dye /probe, intravital microscopy, bioimaging /biomedical imaging, laboratory mouse
GDNF AND SYNAPTIC COMPETITION AND MAINTENANCE
William D Snider, Director, Neuroscience Center
University Of North Carolina Chapel Hill Office Of Sponsored Research Chapel Hill, Nc 27599
Grant 4R37NS037873-06 from National Institute Of Neurological Disorders And Stroke IRG: NSS
Project start date: 1998-07-06
Project end date: 2005-04-30
4R37NS037873-06 (2003): $417694
Core--Multiphoton And Confocal Imaging
William D Snider, Director, Neuroscience Center
University Of North Carolina Chapel Hill Office Of Sponsored Research Chapel Hill, Nc 27599
Grant 1P30NS045892-019005 from National Institute Of Neurological Disorders And Stroke IRG: NSD
Keywords: biomedical facility, neuroimaging, confocal scanning microscopy, developmental neurobiology, fluorescence microscopy, fluorescent dye /probe, intravital microscopy, bioimaging /biomedical imaging, laboratory mouse
Project start date: 2003-07-01
Project end date: 2008-06-30
William D Snider, Director, Neuroscience Center
University Of North Carolina Chapel Hill Office Of Sponsored Research Chapel Hill, Nc 27599
Grant 1P30NS045892-019002 from National Institute Of Neurological Disorders And Stroke IRG: NSD
Keywords: biomedical facility, gene expression, in situ hybridization, neurogenetics, nucleic acid probe
Project start date: 2003-07-01
Project end date: 2008-06-30