1985 |
Tabakoff, Boris |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Ethanol Tolerance--the Study of Its Determinants @ University of Illinois At Chicago
It has been proposed that the effects of ethanol in the CNS are mediated by perturbation of neuronal membrane lipid structure, and that resistance of the membranes to ethanol's effects is involved in ethanol tolerance. The function of enzymes embedded in cell membranes is influenced by the surrounding lipids, and measurement of the temperature dependence of enzyme activity (Arrhenius plots) is thought to provide an indication of the physical state of neuronal membrane lipids. We have demonstrated that, in ethanol-treated animals, there is a change in the lipid-dependent properties of neuronal Na+, K+ ATPase. In order to determine whether this change is related to ethanol tolerance, we now propose to measure enzyme activity during the acquisition and dissipation of tolerance in several strains of mice (which acquire tolerance of different rates). Furthermore, we will alter the rates of acquisition and dissipation of tolerance with neurotoxin and vasopressin treatment, and assess lipid-dependent enzyme activity. If the changes in neuronal membrane properties reflected in enzyme activity are determinants of tolerance, then their appearance and disappearance should parallel those of behavioral tolerance. The activity of two enzymes, Na+ K+ ATPase, and 5'-necleotidase, which are located in different areas of the neuronal membrane, will be assessed. To further determine the importance of lipid properties in the observed changes in enzyme activity, delipidation and reconstitution of Na+, K+ ATPase activity from control and ethanol-tolerant animals will be performed.
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0.936 |
1990 — 1992 |
Tabakoff, Boris |
T32Activity Code Description: To enable institutions to make National Research Service Awards to individuals selected by them for predoctoral and postdoctoral research training in specified shortage areas. |
Pharmacology @ University of Colorado Denver |
1 |
1991 — 2000 |
Tabakoff, Boris |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Alcohol and Neuronal Signal Transduction @ University of Colorado Denver |
1 |
1993 — 2005 |
Tabakoff, Boris |
T32Activity Code Description: To enable institutions to make National Research Service Awards to individuals selected by them for predoctoral and postdoctoral research training in specified shortage areas. |
Predoctoral Training Program in Pharmacology @ University of Colorado Denver |
1 |
1993 — 1997 |
Tabakoff, Boris |
P50Activity Code Description: To support any part of the full range of research and development from very basic to clinical; may involve ancillary supportive activities such as protracted patient care necessary to the primary research or R&D effort. The spectrum of activities comprises a multidisciplinary attack on a specific disease entity or biomedical problem area. These grants differ from program project grants in that they are usually developed in response to an announcement of the programmatic needs of an Institute or Division and subsequently receive continuous attention from its staff. Centers may also serve as regional or national resources for special research purposes. |
Role of Neuronal Systems in Ethanol Withdrawal Seizures @ University of Colorado Denver
Physical dependence on ethanol may be defined by the appearance of withdrawal symptoms upon cessation of ethanol intake. Changes in the function of three neuronal systems have been implicated in the generation or expression of one of these symptoms, withdrawal seizures: reduction in activity of the GABAA receptor, and increased activity of dihydropyridine-sensitive voltage-dependent calcium channels and NMDA receptor-coupled ion channels. Some evidence for the role of these systems in ethanol withdrawal is derived from lines of mice that have been selectively bred for sensitivity (withdrawal seizure prone, WSP) or resistance (withdrawal seizure resistant, WSR) to ethanol withdrawal seizures. WSP mice have more hippocampal NMDA receptors than WSR mice, both before and after chronic ethanol ingestion, and voltage-sensitive calcium channels are increased more in WSP mice after chronic ethanol treatment. WSP mice are also more sensitive to GABA antagonist-induced seizures than WSR mice, and recent evidence suggests a difference in GABAA receptor structure between these lines. We propose to study mice being generated in a new selection study for sensitivity (HW mice) and resistance (LW mice) to ethanol withdrawal seizures. We will assess the sensitivity of the untreated and ethanol-exposed mice, over several generations, to the convulsant properties of drugs acting at the three neurochemical systems of interest (i.e., picrotoxinin, NMDA, Bay K 8644); the measure used will be handling-induced convulsions, which is the selection measure. We will also characterize the NMDA receptor complex and the voltage-sensitive calcium channel in brain by membrane binding and autoradiography. Replication of earlier receptor binding results in an independent selection study will significantly strengthen the relationship between a particular neurochemical system and the generation of withdrawal seizures. This study also allows concurrent behavioral and neurochemical analyses as the selected lines are separating, strengthening the causal relationship. Studies are also proposed to assess interactions between the NMDA and GABAA receptor complexes that may contribute to ethanol withdrawal seizures. Acutely, ethanol inhibits NMDA receptor function and chronic ethanol in vivo as well as chronic exposure of cells in vitro to the NMDA antagonist MK-801 lower the levels of mRNA for certain subunits of the GABAA receptor. We will use primary cultures of cerebellar granule cells to determine if ethanol and NMDA antagonists similarly affect GABAA receptor function, subunit mRNA levels and subunit proteins, to evaluate the hypothesis that a primary effect of ethanol on NMDA receptor function leads to altered GABAA receptor function. Ethanol withdrawal has been likened to kindling, and an understanding of the mechanisms of ethanol withdrawal seizures may lead to the development of efficacious therapies to reduce these seizures and prevent the development of severe withdrawal symptoms in chronic alcoholics.
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1 |
2001 — 2005 |
Tabakoff, Boris |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. R24Activity Code Description: Undocumented code - click on the grant title for more information. |
Gene Array Technology Center For Alcohol Research @ University of Colorado Denver
APPLICANT?S ABSTRACT: The aim of this proposal is to establish a gene array technology core facility (the Gene Array Technology Center for Alcohol Research, GATC) to serve as a national resource for alcohol research. The collective goal of the GATC is to provide the technology base to allow investigators to delineate the gene expression profiles for ethanol-induced changes in transcriptional responses and to delineate inherent differences in gene expression profiles in animals (including humans) which display a particular type of response to ethanol. The GATC will format and print on glass slides a standardized set of gene arrays for human, mouse, and rat. These arrays will be available to NIAAA investigators for use in their alcohol- related research. The GATC will be an invaluable resource that will provide all NIAAA investigators access to gene expression discovery technologies. The GATC website will provide detailed protocols for isolation of RNA, preparation of fluorescently-labeled target probes, and hybridization procedures. The website will be interactive for GATC consultation- with investigators having queries regarding experimental procedures, quality control, and troubleshooting. For investigators that do not have array scanner access, hybridized slides may be sent back to the GATC, where they will be analyzed. The data will be returned to the investigator in spreadsheet form. The GATC bioinfonnaties group will work with NIAAA investigators for analysis of their gene expression data. The bioinformatics group will also develop a highly integrated database that includes gene expression profile data with DNA sequence from experiment protocols involving ethanol- induced changes in transcriptional responses and differences in gene expression profiles inherent to specific responses to ethanol. When NLAAA investigators publish their gene expression studies, the findings will be integrated into the NIAAA gene expression database for alcohol research. The GATC is also prepared to provide education and training for gene array technology, bioinformatics, and database management. A "visiting scientist" program will be established where small groups of 3-10 NIAAA investigators will work in the GATC during intensive three-day workshops. Finally, the GATC will define normal gene expression patterns in specific regions of the brain and the liver, two primary tissues affected by alcohol. To understand "abnormal" gene expression in alcohol responses, "normal" gene expression must be defined. ne analysis of normal gene expression will be a major database resource for NLKAA investigators. Cumulatively, the GATC will provide the gene arrays, informatics power, and database management for gene expression discovery that will form the basis for new hypotheses in alcohol research.
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1 |
2001 — 2006 |
Tabakoff, Boris |
U01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Pathways to Alcohol Tolerance @ University of Colorado Denver
This proposal is directed at determining the molecular mechanisms by which tolerance develops to anxiolytic actions of ethanol. The understanding of the mechanisms of tolerance development will provide information that should assist in the understanding of the motivation driving increased ingestion of alcohol. Our research design will utilize genetically-modified animals which carry a modification in an enzyme in their brains through which alcohol can exert its action (i.e., adenylyl cyclase). The transgenic mice and their "wild-type" littermates will be treated acutely and chronically with ethanol. We will study the effect of drug administration on the animals' levels of anxiety and changes in drug response over a period of chronic drug administration. We will also monitor environmental influences on drug response after chronic ethanol treatment. Simultaneously, we will monitor the function and changes in function of intraneuronal signalling systems that control gene transcription. Finally, we will use gene- expression arrays to monitor which genes are modulated in the short- term and long-term by ethanol administration and which genes are related to tolerance development. In all, our studies should provide an excellent foundation for understanding the neuroadaptive processes leading to ethanol tolerance in reference to the anxiolytic action of this drug.
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1 |
2006 — 2010 |
Tabakoff, Boris |
U01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Colorado Gene Array Core @ University of Colorado Denver
[unreadable] DESCRIPTION (provided by applicant): The Colorado Gene Array Core will focus on the creation of databases that can be used for developing expression QTL (eQTL) maps for mouse and rat brain. We are currently expanding the database of brain gene expression profiles for males and females of the BxD Rl mouse strains. The proposed studies will generate a similar database of gene expression profiles from brains of 31 Rl strains of BxH/HxB rats and the progenitor strains (males and females separately), and microsatellite and SNP genotyping of the rat strains will be performed in collaboration with Dr. Rob Williams, University of Tennessee. This information will allow for eQTL mapping for rat brain gene expression as well as mapping of QTLs for alcohol drinking in the rat strains. When combined with QTL analysis of complex traits, the generation of eQTLs facilitates the identification of candidate genes for these traits, and the Core will use this procedure and the generated data to identify candidate genes that contribute to anxiety and other alcohol related behaviors in the rats. The availability of brain gene expression data and eQTL mapping of the mouse and rat brain transcriptome will produce a novel and unique resource for use by alcoholism investigators. The Core will also perform gene expression analysis and quantitative RT-PCR for collaborating INIA investigators, and provide access to methodology for normalization, statistical analysis and eQTL searches of the generated data. [unreadable] [unreadable] [unreadable]
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1 |
2007 — 2011 |
Tabakoff, Boris |
R24Activity Code Description: Undocumented code - click on the grant title for more information. |
Gene Array Technology Center For Alcohol Research (the R-Gap) @ University of Colorado Denver
DESCRIPTION (provided by applicant): The Rodent Gene Array Program (R-GAP) will focus on the creation of databases that can be used for developing expression QTL (eQTL) maps for mouse and rat brain and for use in extensive studies of expression control elements. The proposed studies will generate a database of gene expression profiles from brains of 75 Rl strains of ISSXILS mice (LXS mice;males and females separately), and integrate ultrahigh density SNP genotyping of the mouse strains. Additionally, in collaboration with the Gene Network, a web-based resource for gene expression analysis, all of our key data for LXS strains will be curated and made available for public use. These valuable new datasets will catalyze eQTL mapping, as well as mapping of QTLs for alcohol-related behaviors currently being acquired in the LXS set. When combined with QTL analysis of complex traits, the generation of eQTLs facilitates the identification of candidate genes for these traits, and R-GAP will exploit this strategy to identify strong candidate genes that contribute to anxiety and alcohol related behaviors in LXS mice, as well as in other strains of mice and rats. The availability of brain gene expression data and eQTL mapping of brain transcriptome will produce a novel and unique resource for use by alcoholism investigators. The data on gene expression which we will derive from large panels of inbred mice will complement data generated from the Rl strains of mice, and is structured to take advantage of whole genome sequencing data being generated by the NIEHS Mouse Resequencing Project. The RGAP database will also feature an integrated neuroinformatics component that provides information and methods that can be used to search for 5'transcriptional control modules that interact with transcription factors located within the eQTLs and epistatic loci. The R-GAP will not only generate data valuable to alcoholism investigators, but it will also provide access to methodology for normalization, statistical analysis and eQTL searches of the generated data. R-GAP will, in essence, act as the "back end" of a data generating and initial screening component which will be used with the GeneNetwork. GeneNetwork will generate the "front end" features to make all data available to the research community at large.
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1 |
2011 — 2013 |
Hoffman, Paula Tabakoff, Boris |
U01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Regulation of Alcohol Consumption by Chromatin Modification @ University of Colorado Denver
DESCRIPTION (provided by applicant): During the previous INIA (Integrated Neurosciences Initiative on Alcoholism)-West project period a genetical genomic/phenomic approach was applied to identify candidate genes in rodents that, through variation in expression levels, contribute to the predisposition to consume varying amounts of alcohol. One such candidate gene is Gnb1 (the G protein beta 1 subunit, Gpi). Brain levels of the Gpi protein are inversely correlated with levels of alcohol consumption in inbred and recombinant inbred strains of mice and in lines of mice selected for differences in alcohol consumption. Treatment of mice with a lentiviral vector expressing a shRNA targeting Gnb1 lowered Gpi levels in nucleus accumbens and increased alcohol consumption by DBA/2 mice. Gpi, as a dimer with Gy proteins, affects numerous intracellular signaling pathways, and has also been reported to interact with, and modify the transcriptional activity of, the Class lla histone deacetylases (HDACs) 4 and 5. HDACs generally repress transcription via modification of histone proteins and class lla HDACs can shuttle between nucleus and cytoplasm, which can regulate their transcriptional activity. In particular, the direct Gpy-HDAC interaction reduces HDAC activity, and therefore has the potential to regulate the transcription of other genes. Given that HDACS activity has previously been found to be crucial for the rewarding effect of cocaine, and that HDAC inhibition reduces alcohol consumption, a role for the Gpy-HDAC interaction in alcohol consumption is postulated. Alcohol consumption will be measured after lowering Gnb1 expression in brains of low alcohol-consuming mice, and after inhibiting or lowering expression of HDACs in brain of high alcohol-consuming mice. To assess the link between Gpi and HDACs, HDAC transcriptional activity (transcript levels and promoter acetylation), as well as HDAC subcellular localization, will be determined after lowering Gpi with RNAi. Overall, this project will investigate molecular mechanisms, focusing on chromatin modification, by which a previously identified candidate gene for alcohol consumption/preference influences transcriptional networks that may underlie this phenotype.
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1 |
2012 — 2016 |
Hoffman, Paula Saba, Laura Maren (co-PI) [⬀] Tabakoff, Boris |
R24Activity Code Description: Undocumented code - click on the grant title for more information. |
Rgap: the Heritable Transcriptome and Alcoholism @ University of Colorado Denver
DESCRIPTION (provided by applicant): This is a renewal application to continue and expand a resource for systems genetic analyses of the rodent (mouse and rat) transcriptome. We have created a website, http://Phenogen.ucdenver.edu, that makes available high quality genetic and whole genome microarray expression data (brain and other organs) from recombinant inbred and inbred strains of mice and rats. This website also provides an array of tools for gene expression analysis. The tools allow a user to identify candidate genes and transcriptional networks for complex physiological and behavioral traits based on the tenets of quantitative genetics, i.e., by combining transcript expression and phenotypic data with expression and behavioral QTL data. We now propose to expand our focus on the rat species, a favored model for human disease. We will breed additional strains of the HXB/BXH recombinant inbred rat panel, and add a panel of genetically diverse classical inbred rat strains, to form a hybrid, high-resolution association mapping panel of rats. Rat strains will be genotyped, and we will complete RNA-Seq analysis of total RNA from brain and liver of all rat strains. We will combine data on exons of protein-coding transcripts with already available exon microarray data, and we will identify, quantify and catalog both protein-coding and non-coding transcripts (including miRNAs, other small non-coding RNAs and long non-coding RNAs) from both organs. We will calculate the heritability of transcript expression levels as one measure of functionality. We will use quantitative data on the transcripts to identify organ-specific genetic locations of transcriptional control by performing association (eQTL) analysis using genetic marker information from each strain. The combined genotypic and transcript expression data will be incorporated into coexpression network modules and we will calculate module QTLs. The module analysis will also provide means to assign gene expression in a whole organ to cell types and anatomical regions of an organ. The annotated and curated data and systems genetic tools will be made available to investigators on the PhenoGen website. We will measure alcohol consumption and alcohol metabolism in the hybrid rat panel, and will integrate the genetic, transcriptome and transcriptional network data with information on these phenotypes to generate causal networks that elucidate the genetic, epigenetic and genomic contributions to predisposition to phenotypic variability.
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1 |
2017 — 2021 |
Hoffman, Paula Saba, Laura Maren (co-PI) [⬀] Tabakoff, Boris |
R24Activity Code Description: Undocumented code - click on the grant title for more information. |
The Heritable Transcriptome and Alcoholism @ University of Colorado Denver
The goal of this application is to establish an animal model and accompanying database suitable for a systems genetic analysis of complex traits, specifically traits that represent genetic predisposing factors for alcohol use disorder (AUD). Systems genetic approaches require a global analysis of factors such as gene expression, protein and metabolite levels in multiple tissues of an organism, as well as an understanding of gene-gene and gene-environment interactions, and the interdependency of these factors in contributing to complex traits/disorders. As a result, a key requirement for an animal model is a genetically stable population that can be studied repeatedly, over many generations, to provide cumulative data that can eventually allow for a complete systems genetic analysis. During the past grant periods, we have progressed well with the development of a Hybrid Rat Diversity Panel (HRDP) that meets these criteria. We have chosen to focus on the rat, rather than the mouse, for studies of complex traits related to AUD, because of the greater size of the rat brain, the ease of training in operant tasks, and the rat?s higher cognitive ability. We have generated DNA sequence data, RNA sequence data and whole genome exon array data on four tissues (brain, liver [whole organ and cell-specific data], heart and brown adipose tissue) from rat strains of the HXB/BXH recombinant inbred (RI) panel and from classic inbred rat strains. We have mapped QTLs for behavioral/physiological traits (alcohol consumption, alcohol deprivation effect, alcohol metabolism including acetate levels after alcohol administration), as well as used transcriptome data to map expression QTLs, to generate transcript coexpression modules and map module eigengene (first principal component) QTLs. These data have been used to identify candidate genes and transcriptional networks that contribute to the measured biochemistry and behaviors. All of our raw, processed and analyzed data have been made available to the research community on our PhenoGen website (http://phenogen.ucdenver.edu). This website, that we developed, also includes several visualization tools to explore these data in a systems genetics framework and allows the user to observe genetic relationships between a complex phenotype of interest and networks of gene products that influence the phenotype. We are now proposing to complete the main core of transcriptional data for the 96- strain HRDP, adding data from another rat RI panel (FXLE/LEXF) and more inbred rat strains. We will obtain full transcriptome information of brain and liver of male and female rats from all strains, quantify the expression of transcript isoforms, including 3?UTR isoforms, and analyze the 3?UTR regions for alternative use of polyadenylation sites and miRNA binding sites. We will use our established and newly developed pipelines to disseminate integrated, systems level data (PhenoGen and Rat Genome Database). We will also expand our demonstration for applying the gathered information to the identification of genetic factors that are linked to the development of AUD by obtaining information on predisposition to ?depression? in the HXB/BXH RI panel, and we will continue to integrate the animal data with human GWAS data.
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1 |