TRANSCRIPTION FACTOR/REGULATION IN LEARNING AND MEMORY
Sandra Pena de ortiz
University Of Puerto Rico Rio Piedras Box 21790 San Juan, Pr 009311790
Grant 5S06GM008102-300076 from National Institute Of General Medical Sciences IRG: MPRC
Abstract: Adapted from Application) The overall goal of this research is to examine the role of transcriptional gene regulation in acquisition and retention of spatial memory. Spatial memory is associative in nature and relates to the orientation and movement of an individual in space in relation to the location of objects in the surrounding environment. A basic understanding of how spatial memories are formed will enable the design of appropriate and effective treatment and prevention strategies for the various neurologic and neuropsychiatric conditions that display alterations in memory processing, such as Alzheimer s disease (AD) and stroke. Regulation of gene expression is considered essential for the establishment of long-lasting cellular changes that underlie the formation of long-term memory. The hzf-3/nurr1 gene is a member of the family of inducible orphan nuclear receptors (ONRs), comprised by immediate-early transcription factors thought to be involved in synaptic plasticity. Recent studies by the PI have shown that the mRNA levels of hzf-3 increase in the rat hippocampus following induction of long-term potentiation at the hippocampal mossy fiber-CA3 synapse and during acquisition of the hole board food search task for spatial memory. Results suggest that the expression of the hzf-3 gene in the hippocampus is closely associated with acquisition and retention of spatial learning. The investigators hypothesize that the hzf-3 protein product is required for normal learning acquisition and that it regulates the transcription of genes whose control is important for appropriate spatial memory formation. This proposal will examine the role of the hzf-3/nurr1 protein product in the formation of long-term spatial memory and will identify genes expressed during acquisition and retention of spatial memory including those that require the hzf-3 product for transcriptional activation. Specific aim 1 will examine the effects of sequence specific antisense oligodeoxynucleotides, designed to block translation of the hzf-3 protein product, on the acquisition of the hole board food search task for spatial memory. Oligodeoxynucleotides are microinfused intrahippocampally prior to training in the hole board food search task and the effects on acquisition and retention are determined. The effectiveness of translational disruption of the hzf-3/nurr1 gene will be determined with Western blots and immunohistochemistry and compared with the behavioral effects of the hzf-3 antisense oligodeoxynucleotides. In specific aim 2, cDNA microarrays are used to define a transcripts expression profile of spatial learning in the Hole Board Food Search Task. Commercially available cDNA microarrays used initially to select candidate learning and memory transcripts will be used to prepare macroarrays on glass slides that will be used for further analysis and validation. The arrays are hybridized with rat hippocampal cDNA probes prepared from rats trained in the food search maze. Finally, studies in specific aim 3 will define an hzf-3 dependent transcriptional profile of spatial learning by combining antisense treatments with cDNA microarray analysis. DNA arrays will be hybridized with cDNA probes prepared from animals trained in the maze after hippocampal injection of either saline, hzf-3 antisense or control oligodeoxynucleotides. The proposed set of experiments will provide new and fundamental knowledge which concerns the molecular mechanisms subserving information storage in the brain and which is necessary for the development of proper management strategies of neural conditions that display problems in memory processing.
Keywords: gene expression, gene induction /repression, learning, long term memory, memory, regulatory gene, space perception, transcription factor, body movement, genetic regulation, nuclear receptor, orientation, antisense nucleic acid, behavioral /social science research tag, immunocytochemistry, laboratory rat, microarray technology, nucleic acid probe, oligonucleotide, western blotting
Sponsored Links Excellgen http://Excellgen.com
Brain Recombination Processes In Learning And Memory
Sandra Pena de ortiz
University Of Puerto Rico Rio Piedras
box 21790
san Juan, Pr 009311790
Grant 5S06GM008102-360085 from National Institute Of General Medical Sciences IRG: ZGM1
Abstract: The overall goal of this proposal is to assess the importance of DNA recombination and DNA repair processes in brain plasticity and complex behaviors. Long-term memory is known to require the production of new proteins in the brain via the regulation of gene transcription and translation mechanisms. Additionally, several reports have postulated the idea that regulation of gene expression and function by DNA recombination mechanisms may also be involved in learning and memory processes. DNA repair mechanisms are tightly associated to recombination processes, such as those utilized in the immune system, which involve DNA cutting, repair, and rejoining (or ligation) mechanisms. Our preliminary studies with a drug that blocks the repair and rejoining of DNA, known as ara-CTP, strongly implicate these processes in the formation of long-term memory in the brain. The experiments proposed here will test specific hypotheses regarding the functional significance of DNA recombination and repair mechanisms in learning and memory processes of the brain. In Specific Aim 1 we hypothesize that ara-CTP impairs memory formation in hippocampal and amygdala dependent behavioral tasks. Just prior to training, ara-CTP, its precursor agent ara-C, or control solution, are injected directly into the brain of rats in order to reach important learning and memory structures. Learning and long-term memory is assessed in rats. Biochemical processes subserving the effects of ara-CTP will also be studied. In Specific Aim 2 we explore the hypothesis that blockade of memory by ara-C is due to an impairment in the regulation of genes important for learning and memory that might be directly or indirectly regulated by DNA recombination in the brain. For these studies we combine the behavioral experiments with gene expression profiling with microarrays. Results from this work will provide fundamental knowledge concerning how the brain stores information, which may help understand the pathophysiology of neurodegenerative and psychiatric diseases, such as Alzheimer´s disease and post-traumatic stress syndrome
Keywords: DNA repair, brain, genetic recombination, learning, memory, molecular psychobiology, neural plasticity, neurogenetics, neuroregulation amygdala, cytosine analog, gene expression, genetic regulation, hippocampus, inhibitor /antagonist, neuropsychology behavior test, behavioral /social science research tag, laboratory rat, microarray technology
TRANSCRIPTION FACTOR/REGULATION IN LEARNING AND MEMORY
Sandra Pena de ortiz
University Of Puerto Rico Rio Piedras Box 21790 San Juan, Pr 009311790
Grant 5S06GM008102-320076 from National Institute Of General Medical Sciences IRG: MPRC
Keywords: gene expression, gene induction /repression, learning, long term memory, memory, regulatory gene, space perception, transcription factor, body movement, genetic regulation, nuclear receptor, orientation, antisense nucleic acid, behavioral /social science research tag, immunocytochemistry, laboratory rat, microarray technology, nucleic acid probe, oligonucleotide, western blotting
Grants awarded to Sandra Pena de ortiz
ADVANCING COMPETITIVE BIOMEDICAL RESEARCH IN PUERTO RICO
Sandra Pena de ortiz
University Of Puerto Rico, Office Of The President, San Juan, Pr 00926-1117
Grant 5P20RR016470-10 from National Center For Research Resources
Abstract: This renewal of PRAABRE represents a step forward in our efforts to foster the continued development of the biomedical research infrastructure in Puerto Rico. Such continued development is achieved through the implementation of a strengthened and cohesive structure with improved integration of common scientific and educational interests, collaborations, and a newly created Mentoring Initiative. The structure of PRAABRE is represented in the following components Administrative, Bioinformatics, Scientific Network, Centralize Research and Instrumentation, and Science and Technology and Education. The Administrative Core provides the programmatic direction for the entire PRAABRE. The new Mentoring Initiative will standardize mentoring practices among participating Institutions at all levels of intervention, from undergraduate student research to mentored research at the faculty level. This initiative will also coordinate Grantsmanship Workshops offered by Kentucky INBRE personnel on a yearly basis. The Administrative Core, via a close interaction with the External Advisory Committee and an External Independent Evaluator, will also organize the overall formative and additive evaluation process of PRAABRE. The Bioinformatics Core will assertively foster and stimulate the development of bioinformatics research, collaborations, and consulting services. This core will also be responsible for maintaining high performance lnternet-2 network capabilities at participating Institutions. Our initiative for providing Access to biomedical Research Electronic Information will also be coordinated by the Bioinformatics Core. The Scientific Network will host 11 INBRE sponsored research projects in PRAABRE, all organized into thrust areas within the biomedical sciences. The Centralized Research and Instrumentation Core integrates the corresponding facilities established at the UPR-Rio Piedras and Medical Sciences Campuses by prior BRIN and INBRE cycles, as well as by COBRE and RCMI, all NCRR programs. The Science and Technology Competence and Education Core will host the PRAABRE Seminar Series, Technology Transfer Awards, Faculty Summer Internships, Continued Education and Technology Competence Workshops, and the Science-on-Wheels project, a K-12 initiative. This PRAABRE Core will also help coordinate Summer Internships in Kentucky for PRAABRE students, an Initiative covered by the Kentucky INBRE. A special quality of this renewal application is the collaborations among the distinct cores, which makes the proposed efforts a true networking of ideas, education, and biomedical research in the Island of Puerto Rico. PUBLIC HEALTH RELEVANCE This renewal application will further augment and strengthen Puerto Rico´s biomedical research capacity, productivity, and competitiveness. PRAABRE will continue to build upon the established biomedical research network of 16 Institutions with a scientific focus on Neuroscience, Molecular Medicine, and Drug Design. PRAABRE will also serve as a pipeline for students throughout the development of their scientific careers and contribute to the development of a knowledge-based economy in the Island
Project start date: 2001-09-15
Project end date: 2014-05-31
Budget start date: 1-JUN-2010
Budget end date: 31-MAY-2011
PFA/PA: PAR-08-150
5P20RR016470-10 (2010): $3518120
2P20RR016470-09 (2009): $2225146
Brain Recombination Processes In Learning And Memory
Sandra Pena de ortiz
University Of Puerto Rico Rio Piedras Box 21790 San Juan, Pr 009311790
Grant 2S06GM008102-330085 from National Institute Of General Medical Sciences IRG: ZGM1
Abstract: The overall goal of this proposal is to assess the importance of DNA recombination and DNA repair processes in brain plasticity and complex behaviors. Long-term memory is known to require the production of new proteins in the brain via the regulation of gene transcription and translation mechanisms. Additionally, several reports have postulated the idea that regulation of gene expression and function by DNA recombination mechanisms may also be involved in learning and memory processes. DNA repair mechanisms are tightly associated to recombination processes, such as those utilized in the immune system, which involve DNA cutting, repair, and rejoining (or ligation) mechanisms. Our preliminary studies with a drug that blocks the repair and rejoining of DNA, known as ara-CTP, strongly implicate these processes in the formation of long-term memory in the brain. The experiments proposed here will test specific hypotheses regarding the functional significance of DNA recombination and repair mechanisms in learning and memory processes of the brain. In Specific Aim 1 we hypothesize that ara-CTP impairs memory formation in hippocampal and amygdala dependent behavioral tasks. Just prior to training, ara-CTP, its precursor agent ara-C, or control solution, are injected directly into the brain of rats in order to reach important learning and memory structures. Learning and long-term memory is assessed in rats. Biochemical processes subserving the effects of ara-CTP will also be studied. In Specific Aim 2 we explore the hypothesis that blockade of memory by ara-C is due to an impairment in the regulation of genes important for learning and memory that might be directly or indirectly regulated by DNA recombination in the brain. For these studies we combine the behavioral experiments with gene expression profiling with microarrays. Results from this work will provide fundamental knowledge concerning how the brain stores information, which may help understand the pathophysiology of neurodegenerative and psychiatric diseases, such as Alzheimer s disease and post-traumatic stress syndrome.
Keywords: DNA repair, brain, genetic recombination, learning, memory, molecular psychobiology, neural plasticity, neurogenetics, neuroregulation, amygdala, cytosine analog, gene expression, genetic regulation, hippocampus, inhibitor /antagonist, neuropsychology, behavior test, behavioral /social science research tag, laboratory rat, microarray technology
Project start date: 2004-07-01
Project end date: 2008-06-30
DNA RECOMBINATION/REPAIR MECHANISMS IN MEMORY
Sandra Pena de ortiz
University Of Puerto Rico Rio Piedras, Box 21790, San Juan, Pr 00931-1790
Grant 5SC1MH086072-03 from National Institute Of General Medical Sciences
Abstract: Does the brain possess a mechanism for generating genetic diversity that could support memory storage? This proposal aims to test the idea that DNA recombination/repair mechanisms play a specific part in memory storage in the brain. Gene rearrangements may be used in the brain as a mechanism to generate experience-dependent protein diversity that could contribute to the storage of information acquired throughout a lifetime. DNA recombination may serve as a mechanism upstream of transcription to regulate the expression and function of a specific set of genes important for long-lasting memory storage. Thus, epigenetic, transcriptional, and recombinational mechanisms of gene regulation in memory do not exclude one another, but most likely complement each other. At this point, the most intriguing questions related to the idea of gene recombination and memory formation in the brain are what are the factors that mediate such a process in neurons and, more importantly, what are the genes that are subjected to this kind of regulation in response to learning. In this proposal, we focus on consolidation mechanisms of conditioned taste aversion (CTA), a behavioral paradigm characterized by the ability of many animals to learn to avoid certain substances after experiencing an unpleasant or harmful somatic (visceral) reaction. Aim # 1 of this proposal addresses the role of amygdalar DNA recombination mechanisms in consolidation of CTA in rats by using or gene knockdown approaches, both targeting the function of putative recombination effector enzymes, Flap Structure-Specific Endonuclease-1 (FEN-1) and DNA ligase IV. AIM # 2 examines amygdalar genomic rearrangement of putative DNA recombination target genes, such as protocadherin p9. Particularly, experiments will determine if the protocadherin 09 gene undergoes CTArelated genomic rearrangement in amygdala neurons. Overall, these studies will establish that DNA recombination/repair processes are part of the initial mechanisms utilized by the brain for the long-lasting storage of information, characterize the function of specific factors involved in this processes, and help demonstrate that specific gene targets undergo genomic rearrangement during memory formation
Keywords: Address; Affect; Ammon Horn; Amygdala; Amygdaloid Body; Amygdaloid Nucleus; Amygdaloid structure; Animals; Assay; Behavioral; Behavioral Paradigm; Binding; Binding (Molecular Function); Bio-Informatics; Bioassay; Biochemical; Bioinformatics; Biologic Assays; Biological Assay; Blotting, Southern; Brain; Brain region; Caudate Nucleus; Caudate nucleus structure; Common Rat Strains; Complement; Complement Proteins; Control Groups; Cornu Ammonis; DNA; DNA Chips; DNA Joinases; DNA Ligases; DNA Microarray; DNA Microarray Chip; DNA Microchips; DNA Rearrangement; DNA Recombination; DNA Restriction Enzymes; DNA Sequence Rearrangement; DNA blotting; DNA ligase IV; DNA recombination (naturally occurring); Data; Deoxyribonucleic Acid; Down-Regulation; Down-Regulation (Physiology); Downregulation; Encephalon; Encephalons; Enzymes; Epigenetic; Epigenetic Change; Epigenetic Mechanism; Epigenetic Process; Event; FEN-1; Fen1 Protein; Flap Endonuclease 1; Flaps; Flavoring; Flavoring Agents; Gene Action Regulation; Gene Expression; Gene Expression Regulation; Gene Rearrangement; Gene Regulation; Gene Regulation Process; Gene Targeting; Gene Transcription; Gene variant; Genes; Genes, RAG-1; Genetic Diversity; Genetic Recombination; Genetic Transcription; Genetic Variation; Genome; Genomics; Gustation; Hippocampus; Hippocampus (Brain); Homologous Recombinational Repair; Immunoglobulin V(D)J Rearrangement; Information Storage; Infusion; Infusion procedures; Injection of therapeutic agent; Injections; Intermediary Metabolism; Island Flaps; Laboratories; Learning; Length of Life; Ligase; Longevity; Lymphocyte; Lymphocytic; METBL; Mammals, Rats; Mammals, Rodents; Mediating; Memory; Metabolic Processes; Metabolism; Methods and Techniques; Methods, Other; Molecular Interaction; NHEJ; Nerve Cells; Nerve Unit; Nervous System, Brain; Neural Cell; Neurocyte; Neurons; Non-Homologous End Joining; Nonhomologous DNA End Joining; Nucleus Caudatus; Oligo; Oligonucleotides; Peptide Signal Sequences; Play; Polydeoxyribonucleotide Ligases; Polydeoxyribonucleotide Synthetases; Polymerase; Process; Proteins; RAD2 Homolog-1 Nuclease; RAG1 gene; RNA Expression; RTH-1 Nuclease; Rat; Rattus; Reaction; Rearrangement; Recombination; Recombination Repair; Recombination, Genetic; Recombination-Activating Genes-1; Regulation; Reporting; Restriction Endonucleases; Rodent; Rodentia; Rodentias; Role; Sampling; Signal Peptide; Signal Sequences; Signal Sequences, Peptide; Site; Southern Blotting; Specificity; Storage, Data; Structure; Surgical Flaps; Synthetases; Targetings, Gene; Taste; Taste Perception; Techniques; Testing; Time; Training; Transcription; Transcription, Genetic; V(D)J Rearrangement; V(D)J Recombination; VDJ rearrangement; VDJ recombination; Validation; Variation (Genetics); Visceral; Work; allelic variant; amygdaloid nuclear complex; cDNA Probes; caudate nucleus; conditioned fear; design; designing; endonuclease; experience; experiment; experimental research; experimental study; fear conditioning; gene discovery; gene product; hippocampal; inhibitor; inhibitor/antagonist; laser capture microdissection; life span; lifespan; long term memory; lymph cell; neuronal; protein signal sequence; recombinational repair; research study; response; social role
Project start date: 2008-09-24
Project end date: 2012-07-31
Budget start date: 1-AUG-2010
Budget end date: 31-JUL-2011
PFA/PA: PAR-06-491
5SC1MH086072-03 (2010): $335250
5SC1MH086072-02 (2009): $335250
1SC1MH086072-01 (2008): $324225
TRANSCRIPTION FACTOR/REGULATION IN LEARNING AND MEMORY
Sandra Pena de ortiz
University Of Puerto Rico Rio Piedras Box 21790 San Juan, Pr 009311790
Grant 2S06GM008102-290076 from National Institute Of General Medical Sciences IRG: MPRC
Abstract: Adapted from Application) The overall goal of this research is to examine the role of transcriptional gene regulation in acquisition and retention of spatial memory. Spatial memory is associative in nature and relates to the orientation and movement of an individual in space in relation to the location of objects in the surrounding environment. A basic understanding of how spatial memories are formed will enable the design of appropriate and effective treatment and prevention strategies for the various neurologic and neuropsychiatric conditions that display alterations in memory processing, such as Alzheimer s disease (AD) and stroke. Regulation of gene expression is considered essential for the establishment of long-lasting cellular changes that underlie the formation of long-term memory. The hzf-3/nurr1 gene is a member of the family of inducible orphan nuclear receptors (ONRs), comprised by immediate-early transcription factors thought to be involved in synaptic plasticity. Recent studies by the PI have shown that the mRNA levels of hzf-3 increase in the rat hippocampus following induction of long-term potentiation at the hippocampal mossy fiber-CA3 synapse and during acquisition of the hole board food search task for spatial memory. Results suggest that the expression of the hzf-3 gene in the hippocampus is closely associated with acquisition and retention of spatial learning. The investigators hypothesize that the hzf-3 protein product is required for normal learning acquisition and that it regulates the transcription of genes whose control is important for appropriate spatial memory formation. This proposal will examine the role of the hzf-3/nurr1 protein product in the formation of long-term spatial memory and will identify genes expressed during acquisition and retention of spatial memory including those that require the hzf-3 product for transcriptional activation. Specific aim 1 will examine the effects of sequence specific antisense oligodeoxynucleotides, designed to block translation of the hzf-3 protein product, on the acquisition of the hole board food search task for spatial memory. Oligodeoxynucleotides are microinfused intrahippocampally prior to training in the hole board food search task and the effects on acquisition and retention are determined. The effectiveness of translational disruption of the hzf-3/nurr1 gene will be determined with Western blots and immunohistochemistry and compared with the behavioral effects of the hzf-3 antisense oligodeoxynucleotides. In specific aim 2, cDNA microarrays are used to define a transcripts expression profile of spatial learning in the Hole Board Food Search Task. Commercially available cDNA microarrays used initially to select candidate learning and memory transcripts will be used to prepare macroarrays on glass slides that will be used for further analysis and validation. The arrays are hybridized with rat hippocampal cDNA probes prepared from rats trained in the food search maze. Finally, studies in specific aim 3 will define an hzf-3 dependent transcriptional profile of spatial learning by combining antisense treatments with cDNA microarray analysis. DNA arrays will be hybridized with cDNA probes prepared from animals trained in the maze after hippocampal injection of either saline, hzf-3 antisense or control oligodeoxynucleotides. The proposed set of experiments will provide new and fundamental knowledge which concerns the molecular mechanisms subserving information storage in the brain and which is necessary for the development of proper management strategies of neural conditions that display problems in memory processing.
Keywords: gene expression, gene induction /repression, learning, long term memory, memory, regulatory gene, space perception, transcription factor, body movement, genetic regulation, nuclear receptor, orientation, antisense nucleic acid, behavioral /social science research tag, immunocytochemistry, laboratory rat, microarray technology, nucleic acid probe, oligonucleotide, western blotting
Project start date: 1988-06-01
Project end date: 2004-06-30
Sponsored Links Excellgen http://Excellgen.com