1985 |
Goldowitz, Daniel |
R23Activity Code Description: Undocumented code - click on the grant title for more information. |
Genetic Determinants of Brain Structure and Function @ Thomas Jefferson University
The reeler (rl) mutant mouse has provided important insights to cortical development and function. This project is designed to elucidate the site(s) of mutant gene acton in the rl mouse, and relate the structural abnormalities of the mutation to behavioral deficits. Three approaches to the problem of where the rl gene acts will be made: 1) We can test whether an alteration of the extracellulr milieu is the means by which the rl gene exerts is action by transplantion of early embryonic rl cerebella or hippocampal formations. 2) Reeler reaction goes both right and left normal chimeric mice will be created to analyze rl gene action relative to cerebellar granule cells. The ichthyosis phenotype of centrally clumped heterochromatin will be used in one component of the chimera to analyze granule cell populations. 3) I will use a new cell marker in chimeras that should label all cell types and allow me to study the site of mutant gene action in other neuronal and supporting cells in rl reaction goes both right and left normal chimeric brains. The varying expression of the rl phenotype inherent in chimeric mice will permit me to correlate performance of hippocampal-specific behavioral tasks (radial arm maze, spontaneous alternation and two-way avoidance) with both the cytoarchitectonics of the hippocampal subfields and patterns of cholinergic innervation and mossy fiber distribution. After behavioral testing, chimeric mice and controls will be perfused and their brains sectioned for the sequential histological processing and analysis of cerebellar granule cells and hippocampal cytoarchitectonics (Nissl stain), hippocampal cholinergic innervation (acetylcholinesterase histochemistry), and mossy fiber distribution (Timm's stain for heavy metals). The three long term goals of this work are to: 1) Use the information gained about the site of rl gene action to better understand and explain development and abnormalities of the mammalian central nervous system, 2) Establish the use of a new cell marker to reliably study the whole range of nervous system cells in chimeras, and 3) Use the mutant reaction goes both right and left normal chimera as a model system to integrate our knowledge of genetic, anatomical and behavioral phenomena.
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0.931 |
1986 — 1988 |
Goldowitz, Daniel |
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. |
Cns Development and Mutant Gene Action @ Thomas Jefferson University
The genetic and epigenetic mechanisms by which the brain develops into a highly ordered structure are largely unknown. The goal of this proposal is to elucidate some of the early postmitotic events in neurogenesis. There are several neurological mouse mutants which are defective in particular aspects of this developmental program. Two such genetic mutants, the weaver (wv) and the reeler (rl), hold forth fascinating information concerning the genetic control over the migratory phase of neurogenesis. These mutations affect the ability of a single cell type (the wv cerebellar granule cells) or virtually all cell types (rl) to migrate and stabilize appropriately. Experimental mouse chimeras provide a direct means to ascertain the target(s) of mutant gene action. Four cell embryos of normal (Mus caroli) and neurological mutant (Mus musculus) mice will be aggregated to form a single chimeric embryo composed of genetically normal and mutant cells. A new cell marking system will be used to identify each cell of the chimeric brain, as genotypically normal or mutant. This new cell marker, which involves the in situ hybridization of a species-specific cDNA probe to mark Mus musculus but not Mus caroli cells, will permit careful light and electron microscopic analyses of each cell's genotype compared to its phenotype. The genotype/phenotype comparisons will define the intrinsic or extrinsic nature of wv and rl mutant gene action relative to all cell types in the cerebellum. This type of information will provide a better understanding of normal brain development and how abnormal development occurs at both the more obvious (e.g., congenital ataxias) and more subtle (e.g., mental retardation) levels of dysgenesis.
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0.931 |
1992 — 1995 |
Goldowitz, Daniel |
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. |
Genetic Control of Retinal Development @ University of Tennessee Health Sci Ctr
Our aim is to study genetic and environmental factors that control the formation of the connection between eye and brain. We are specifically interested in mechanisms that control whether or not retinal axons cross at the chiasm. This is important for at least two reasons: (1) to better understand the development of binocular vision in humans; and (2) to better understand genetic, molecular, and cellular factors that guide the growth of nerve fibers within the central nervous system. We use two complementary approaches to this important problem in visual system development. First, we will take advantage of powerful new genetic and experimental manipulations in mouse embryos: we will make experimental chimeras by combining albino embryos and transgenic embryos that carry human globin or human neurofilament genes. This will allow us to study the mechanism by which the albino mutation in the mouse sharply reduces the number of retinal ganglion cells with uncrossed projections into the optic tracts. Is the misrouting of retinal axons in pigment-deficient mutants due to an intrinsic defect of retinal ganglion cells, or is it secondary to the effects of the mutations on the environment through which retinal axons grow? Our second approach also focuses on the genetic control of axon guidance, but the approach is quite different. Here we plan to pursue genetic analysis of a highly unusual visual system mutation recently discovered in Belgian sheepdogs (Williams, Garraghty, and Goldowitz, 1991). In these mutants, the entire retinal projection is uncrossed. This striking genetic defect is opposite to that seen in albinos, in which an abnormally large number of optic axons cross at the chiasm. These achiasmatic mutants provide an excellent animal model of inherited nystagmus and should provide genuine insight into the molecular and genetic basis of normal and abnormal development of retinal connections. Three lines of research will be pursued to define the genetic basis of this inherited disease (1) cytogenetics, (2) breeding studies of the transmission of the disease, and (3) linkage analysis to map the location of the mutant gene. This work will provide a foundation for the isolation and characterization of the mutant gene and its product.
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0.939 |
1993 — 1995 |
Goldowitz, Daniel |
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. |
Cns Development and Gene Action @ University of Tennessee Health Sci Ctr
The overall objective of these studies is to determine how genes instruct the development of the mammalian CNS. The study of mutations that affect nervous system development offers an ideal vantage point to approach this problem. The weaver mutant mouse is the subject of the present proposal, and two aspects of the weaver gene (wv) will be examined: l) The identity of the weaver gene will be sought using the techniques of chromosomal microdissection, cloning, and cDNA screening. This approach takes advantage of the most recent advances in the developmental and chromosomal bases of the weaver mutation. Additionally, the mapping of the weaver locus will be continued using subspecies crosses and DNA markers to identify restriction fragment length polymorphisms closer to the weaver locus and to create tools important for the identification of the wv gene. 2) A cellular developmental section will characterize a target of the wv gene, the cerebellar cell in normal and mutant brains. Experiments will also address the hypothesis that the weaver mutation disrupts the process of axonal outgrowth which then leads to cell death. We will employ the invivo use of axonal markers to image the dynamics of granule cell axonal growth in normal and mutant cerebella. Ultrastructural studies will help characterize genetically normal and weaver granule cells for a broader diagnosis of cell pathology in the etiology of the weaver phenotype of granule cell death. These studies are aimed at obtaining a comprehensive understanding of a single locus, wv: its impact on various cell types, its localization in the mammalian genome, and its contextual relationship to brain development. In particular, the massive neonatal loss of cerebellar granule cells and the progressive loss of substantia nigra dopaminergic neurons that occurs in weaver is pathologically similar to the loss of these neurons in the human conditions of cerebellar hypoplasia and Parkinson's disease.
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0.939 |
2000 — 2002 |
Goldowitz, Daniel |
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. |
Histological Phenotyping the Mouse Nervous System @ University of Tennessee Health Sci Ctr
DESCRIPTION (Adapted from the applicant's abstract): A group of experienced neuroscientists will closely interact to provide a high throughput analysis of various aspects of brain structure and implied function. These investigators have had, as a whole, a long history in the neuroanatomic analysis of normal and mutant brain structures. They will collectively produce a high quality database, disseminated on the Internet, that will provide an integrated summary of the major aspects of brain organization in a large variety of inbred and recombinant inbred strains of mice. Three timepoints in the life history of the mouse will be assessed: embryonic day 17.5, postnatal day 60-90, and 18-24 months of age. Several features of the proposed neurohistological screen obviate time consuming methods and introduce a great deal of flexibility for phenotype assessment. Using a set of seven straightforward and reproducible staining procedures (cresyl violet, osmium tetroxide, Timm's stain, anti-glia fibrillary acidic protein and anti-NeuN immunohistochemistry, cytochrome oxidase histochemistry, cell birthdating with BrdU immunocytochemistry), the plan is to assess the cytoarchitectonics, myelinated fiber pathways, terminal fields, astroglia and neuronal populations, activity state, and proliferative populations in a extensive series of adult inbred and hybrid mice that are commonly used for neurobiological studies and transgenic experimentation. These seven procedures will produce a detailed and composite view of neural organization that will be assessed at the qualitative and quantitative levels of analysis. For embryonic and aged mice, additional stains and imaging methods will assess various hallmarks of the developing and aged brain. One important endpoint of these efforts will be to develop a relational database that can be used to compare and measure various aspects of nervous system structure. Strain differences can be readily assessed by consulting this online database. The recovery of apparent mutant phenotypes that are reflected in abnormalities in brain structure can be compared to this database. Furthermore, in collaboration with other researchers in the field, these efforts will coordinate a host of phenotypic data such as cell number, nuclear volume and density of neurons, and molecular expression patterns to begin to mine the wealth of data that is inherent in the natural and quantitative variation of traits in inbred strains of mice.
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0.939 |
2000 — 2003 |
Tretiak, Oleh Nissanov, Jonathan Goldowitz, Daniel Williams, Robert [⬀] |
N/AActivity Code Description: No activity code was retrieved: click on the grant title for more information |
Informatics Center For Mouse Neurogenetics @ National Institutes of Health
IBN-0003982 NIH/NSF Human Brain Project PI: Robert W. Williams Informatics Center for Mouse Neurogenetics
The purpose of this project is to develop and exploit a suite of image databases, motorized Internet microscopes, and software to study the genetic basis of structural variation of the mouse CNS. Resources are open to the research community through an integrated web interface at . The focus of this project is to provide collaborative research environments for mapping quantitative trait loci (QTLs). QTL analysis is a burgeoning field that tackles complex biological traits modulated by many genes. Four new resources and technologies will be developed: 1) the Mouse Brain Library, 2) the Internet Microscope System, 3) the NeuroCartographer Project, and 4) the Neurogenetics Tool Box. Achieving the aims of these four projects will catalyze a new era in the structural analysis of the adult mammalian nervous system and will lead to a large number of novel lines of research on the development and aging of the human brain.
This research is funded in part by the National Science Foundation through its participation in the Human Brain Project, which is a Federal interagency research initiative, mandated by Congress as part of the "Decade of the Brain".
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0.912 |
2000 — 2005 |
Goldowitz, Daniel |
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. |
Targeted Mutagenesis of Mouse Genome &Neural Phenotype @ University of Tennessee Health Sci Ctr
DESCRIPTION (ADAPTED FROM THE INVESTIGATOR'S ABSTRACT): Researchers across the State of Tennessee have combined their expertise in ENU mutagenesis of the mouse genome and neurosciences to conduct a concerted effort to mutagenize the mouse genome, screen for deficits in neural function and structure, and thereby lay the basis for a large scale analysis of the functional genomics of the nervous system. A novel mutagenesis program is employed whereby markers (coat color or molecular/PCR-based) identify mice (test class) that potentially carry mutations in specific chromosomal regions. This approach serves as a strong foundation for the genetic dissection of phenotype and genotype relationships in brain by offering an economy of effort and reliability in addition to pre-localizing mutations to discrete regions of the genome. A profile of neural function is obtained from all potentially mutant mice by high throughput screens that examine basic behavior, gross structure, and histology of the nervous system. Four domains (alcohol, drugs of abuse, eye, and social behavior) will phenotype mature mice from each pedigree. A fifth domain (aging) will focus on late onset phenotypic abnormalities of nervous system function and structure. The use of markers to identify test class animals permits the efficient aging of all pedigrees. Animals that are flagged by demonstrating aberrant results in a primary screen will be moved into secondary screens that characterizes the molecular phenotype of the brain and eye as well as explore performance in the domains of learning and memory, audition, and nociception. The BioInformatics Core tracks all mice, collects and stores the phenotypic data on each, mouse, flags mice with aberrant phenotypes, and provides a means for investigators (both inside and outside the consortium) to access and analyze this data. A Research Community Core will work with out veterinarians to promote the use of our mutant mice by outside researchers. An external advisory panel assists in this effort as well as in the design of additional phenotypic screens.
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0.939 |
2002 — 2006 |
Goldowitz, Daniel |
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. |
Inia: Genetic Analysis of Alcohol Consumption and Stress @ University of Tennessee Health Sci Ctr
DESCRIPTION (provided by applicant): The genetic basis of the neuroadaptive response to stress and anxiety relative to alcohol abuse is poorly understood. In this Research Component of the INIA, a wide net will be cast to identify genes important to neuroadaptation and alcohol abuse by using ethyl nitrosourea (ENU)-induced mutations of male mouse germ cells and subsequent breeding to screen for dominant and homozygous recessive mutations that result in aberrant alcohol-related phenotypes. Microarray and gene mapping studies are proposed to chart the molecular pathways and the specific genes that are involved in alcohol-related phenotypes. In addition, specially constructed congenic lines of mice will be used as reagents to better map loci responsible for quantitative trait loci (QTLS) and serve as reagents for gene identification in an ENU-mutagenesis program. In Aim, 1, ongoing NIH-supported mutagenesis program will be identifying mutants with abnormal fear conditioning behavior and a wide range of aberrant alcohol phenotypes and these mutants will be further explored for a) corticosterone levels following acute ethanol administration and b) stress-induced reinstatement of ethanol consumption. A concerted effort will be made in Aim 2 to identify the genetic basis of quantitative trait (QTLS) for alcohol-related behaviors by targeted mutagenesis of mouse chromosomes 1 and 4. Male congenic or consomic mice that span the withdrawal and drinking QTLs on Chr 1 and 4 will mutagenized with ENU. Test class mice, identified by molecular markers, will be phenotyped for 2-bottle choice, withdrawal or corticosterone levels following ethanol administration. Mice that show aberrant behavior in any of these tasks would be candidates to bear a mutation in a gene responsible for the relevant QTL. These studies should provide insights into the genetic bases of stress-alcohol interactions, and provide molecular clues to rational therapeutic approaches to curing alcoholism.
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0.939 |
2003 — 2005 |
Goldowitz, Daniel |
R25Activity Code Description: For support to develop and/or implement a program as it relates to a category in one or more of the areas of education, information, training, technical assistance, coordination, or evaluation. |
A Neuromutagenesis Training and Education Program @ University of Tennessee Health Sci Ctr
DESCRIPTION (provided by applicant): The NIH has placed a priority on discovering the function of genes identified through the human genome project. The mouse has been recognized as the pre-eminent organism to model human disease and various initiatives have focused on its use in this regard. One powerful means to illuminate gene function is through mutagenesis, and the NIMH and other neurological institutes have funded three centers to carry out chemical mutagenesis in the mouse and screen mice for neuroscience-related phenotypes. To make these centers of core use to the neuroscience community, each center must attract researchers to use the mutagenesis facilities for their research programs. The Tennessee Mouse Genome Consortium (TMGC), a group of eight institutions across the State of Tennessee, is one of the three funded centers and proposes a plan to recruit researchers and students to maximize the potential of our neuromutagenesis program and train individuals in the use of mutant mice for neuroscientific endeavors. We have three components of our training and education plan. First, we will actively work to recruit investigators who demonstrate an abiding interest in applying new phenotyping protocols to our mutant mice. This may represent a new direction for some or a realization of the power of genetic analysis for others, but in all cases we will work with them to understand the basics of our program and to obtain sustained funding for these efforts. Second, we will establish a training program for undergraduate and graduate students and postdoctoral fellows in the behavioral analysis of mutant mice using the TMGC neuromutagenesis program. Finally, we propose to offer an annual workshop-style course in neurogenetics and analysis of mouse behavior. Through these three mechanisms, we plan to create a new pool of researchers that understand the power of neurogenetic approaches and can take advantage of neurological mutant mice in understanding brain structure and function.
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0.939 |
2005 — 2006 |
Goldowitz, Daniel |
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. |
Mapping Cerebellar Development in Time and Space @ University of Tennessee Health Sci Ctr
DESCRIPTION (provided by applicant): The cerebellum is emerging as an important brain region for the coordination of motor and cognitive behaviors. Developmental abnormalities of the cerebellum have been linked to autism, schizophrenia, and other disorders of human neural function. This grant proposes to acquire extensive new data sets for gene expression and cellular phenotypes over six epochs of cerebellar development in over 30 recombinant inbred (RI) strains of mice (BXD) and 15 single gene mutant mice. This data will be web-accessible via WebQTL. We will also develop and integrate web-based informatic and visualization tools for researchers to analyze our data sets, provide datasets of their own for analysis, and test hypotheses about the cellular and molecular development of the cerebellum. Four specific aims are proposed that will be supported by four core functions. In Aim 1, we will obtain the phenotypic data on the full spectrum of expressed genes and several quantifiable developmental processes in RI and mutant mice. This data will be integrated into a current database that houses an exceptional array of phenotypic data on the adult mouse brain, WebQTL. In Aim 2, we will use WebQTL, Bayesian method analysis, graph theoretical approaches to the identification of cliques in expression data, and latent semantic indexing of Medline references to mine data on the patterns, both in time and space, of expressed genes and cellular phenotypes. In Aim 3, we will use molecular (qRT-PCR), anatomical (in situ hybridization and immunocytochemistry) and experimental (siRNA) approaches to validate inferences about gene and phenotype relationships. Finally, in Aim 4, we will develop a web-accessible, 3D, high-end animation of the developing cerebellum that will be used for heuristic and experimental purposes. The data that is obtained and the tools that are constructed in this project will be fully open to the research community. This project is also designed to interface with several of the currently funded Human Brain Projects that look at the anatomy and cell biology of the adult mouse brain and cerebellum. The phenotypic data that is gathered will contribute to the growing understanding of the molecular and cellular bases of cerebellar development. Such information may help understand and treat disorders of cerebellar origin, such as the most common form of childhood brain cancer, the medullablastoma, which is believed to emanate from the developing granule cells of the cerebellum. In the long term, we hope to use the tools developed in this project to make predictions about the molecular pathways and cellular programs that are important to the well-being of the central nervous system
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0.939 |
2006 — 2008 |
Goldowitz, Daniel |
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. |
Gene to Phenotype Networks For Alcohol &Drug Addiction @ University of British Columbia
[unreadable] DESCRIPTION (provided by applicant): Drug abuse and addiction are complex phenotypes. Typical of many human diseases, they are influenced by multiple genetic and environmental factors. Susceptibility to addiction is co-morbid with other behavioral disorders, which is evidence that the same genetic influences may be acting to affect multiple phenotypes, a phenomenon known as gene pleiotropy. The main purpose of this project is to systematically identify genes and gene networks that modulate pleiotropic responses to abused substances, behavioral variation, and susceptibility to abuse. This application exploits the unique mapping properties of Rl strains, a new, high power expanded set of Rl lines, advanced bioinformatics tools, extensive databases present in WebQTL, and the expertise of the TMGC high-throughput phenotyping resource to systematically identify upstream genes and molecular networks that ultimately modulate downstream pleiotropic drug and alcohol phenotypes. The powerful combination of QTL mapping and microarray transcript profiling will be applied to these systems level phenotypes by exploiting existing high-throughput molecular data resources in WebQTL. As part of this application, we have assembled a strong team of investigators with complementary expertise in several areas, most notably in complex trait analysis and gene mapping, behavioral and neural analysis, psychopharmacology and pharmacogenetics, transcriptome profiling and molecular genetics, drug abuse, alcoholism, mouse colony management and distribution and advanced bioinformatics and multivariate statistical methods of handling large data sets. This strong team will capitalize on the generous support offered by the Department of Energy's Oak Ridge National Lab. The data resources generated by this project will dramatically reduce the amount of phenotyping one needs to perform to discover the effects of any novel gene specific mutation. Candidate genes will be validated using a novel banked ENU resource at ORNL as well as publicly available mouse mutant resources. This will be invaluable for the development of realistic complex disease models and will provide data resources to suggest cost effective targeted phenotyping strategies for large scale single gene mutation efforts such as those proposed by the Comprehensive Knockout Mouse Project Consortium. By examining covariance of gene expression measures and known phenotypic measures in BXD Rl lines, we can rationally target phenotypes that are likely to be affected by particular gene mutations. More broadly, we will be able to identify the specific genetic basis of the pleiotropic and polygenic effects of genetic polymorphisms on drug abuse, addiction, and individual differences in brain and behavior. [unreadable] [unreadable] [unreadable]
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1 |
2007 — 2011 |
Goldowitz, Daniel |
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. |
Inia: Mouse Resources Core @ University of British Columbia
DESCRIPTION (provided by applicant): This is a new core to INIA stress that arises out of the generation of many new mouse models through INIA support over the last 4+ years. These mouse models will be important resources to INIA investigators to study the interplay between stress, anxiety and alcohol consumption. With these new resources, we feel it is critical to disseminate them to users and evaluate each of these new mouse models with a set of INIA relevant behavioral analyses to better identify their usefulness to the various researchers in INIA stress and to the alcohol research community and beyond. The final part of this Core will provide key elements to a mouse core: distribution, cryopreservation, genotyping, and a curatorial function to keep an inventory of mouse mutants and inbred lines that we have available to INIA users and to accumulate and annotate the behavioral, cellular, and molecular knowledge about each mutant and inbred line. The specific aims of this Core are as follows: Aim 1 - The maintenance and distribution of novel mutant lines of mice produced by the ENU-mutagenesis program. The production and availability of novel mouse mutants that have abnormal alcohol, stress, and/or anxiety phenotypes represent an exciting resource for multi-disciplinary studies by our INIA stress investigators. This part of the Core will make these lines of mice available to INIA and community-wide researchers. Aim 2 - The maintenance and distribution of the expanded BXD recombinant inbred (Rl) and B6.A consomic lines of mice. This INIA has greatly expanded the phenotypic "space" (molecular, cellular, and behavioral) associated with unique mouse reference populations and this aspect of the Core will make these mice readily available to INIA researchers. This includes the 50 newly developed BXD Rl lines that arose, in part, from the previous INIA support, and the importation of 86.A consomic lines. Aim 3 - The Behavioral Phenotyping component of this Core will provide more comprehensive phenotypic information about EtOH and stress related behaviors in unique genetic mouse models that have been identified by high throughput behavioral screening within the INIA stress Consortium as exhibiting "extreme" phenotypes for EtOH and/or stress/anxiety responsiveness. Aim 4 - Affiliated functions that include curation of mouse lines in the Core, cryopreservation, database management, genotyping, and dissemination of information.
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1 |
2009 — 2013 |
Blaha, Charles (co-PI) [⬀] Goldowitz, Daniel Heck, Detlef H. (co-PI) [⬀] Mittleman, Guy [⬀] |
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. |
Cerebellar Modulation of Frontal Cortical Function
DESCRIPTION (provided by applicant): The developmental loss of cerebellar Purkinje cells that occurs in autism spectrum disorders has been associated with a heterogeneous pattern of cognitive deficits that cannot be explained by a unitary cognitive impairment. It is very unlikely that the simple loss of cerebellar Purkinje cells can directly account for these myriad cognitive deficits. Rather, it is likely that autism is, at its essence, a disconnection syndrome that results, at least in part, from a disruption of cerebellar modulation of the prefrontal cortex (PFC). We have exciting new data suggesting that the cerebellum modulates PFC dopamine levels. Here we propose to investigate the disconnection hypothesis that cerebellar pathology results in dopaminergic abnormalities in the prefrontal cortex (PFC) and underlies some of the core neuropsychiatric symptomatology of autism. In Aim 1 we will determine the pathway(s) whereby the cerebellum modulates dopamine release in the PFC and glutamate release in subnuclei comprising the cerebellum to PFC pathways and the neurochemical, electrophysiological, anatomical, and behavioral consequences of a disconnection between these two structures. Aim 1 will compare wildtype (control) and Lurcher mice that loose all Purkinje cells, to determine the consequences of complete loss of Purkinje cells on cerebellar-PFC communication. Aim 2 will investigate the behavioral and physiological consequences of partial loss of Purkinje cells - as typically found in autistic brains. Using Lurcher-wildtype chimeras with varying developmental loss in Purkinje cell numbers we will determine how neurochemical, electrophysiological, anatomical and behavioral indicators of PFC function depend on Purkinje cell number. Given the well documented reductions in cerebellar neuron number that are found in autism spectrum disorders, the neurochemical, electrophysiological, anatomical and behavioral analyses of chimeric mice presents a unique opportunity to model both the developmental and cerebellar aspects of these syndromes.
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0.964 |
2016 — 2020 |
Goldowitz, Daniel Hamre, Kristin M |
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. |
Maternal Genotype, Choline Intervention,& Epigenetics in Fetal Alcohol Syndrome @ University of Tennessee Health Sci Ctr
? DESCRIPTION (provided by applicant): The type and severity of ethanol-induced alterations following prenatal ethanol exposure is strongly impacted by genetics. However, the role of genetics is complicated by the fact that both the mother and fetus have unique genotypes. The role of genetics could be an even bigger consideration in the evaluation of treatments, since the metabolism of any therapeutic is completely regulated by the mother in early development. This proposal will test the following hypotheses: 1) the genotype of the mother has a significant role in determining the level of ethanol-induced cell death in the developing brain as well as in the efficacy of choline treatment in modulating ethanol-induced cell death and 2) the epigenome of the embryo has an important role in modulating genetic differences in susceptibility to the effects of ethanol exposure. We have been examining the BXD panel of mice, generated by crossing C57BL/6J (B6) and DBA/2J (D2) strains, and show differential sensitivity following equivalent ethanol exposure thereby facilitating the testing of these hypotheses. Specific Aim 1 will test the hypothesis that the maternal genotype influences the level of ethanol-induced cell death in the developing neural tube. Two complementary approaches, reciprocal crosses and embryo transplants, will be used. Embryos will be exposed to ethanol on embryonic day 9 and collected 7 hours after the initial ethanol exposure. Cell death will be quantified from TUNEL and activated caspase-3 labeled sections in the developing telencephalon and brain stem. If, in both approaches, the level of cell death is altered depending upon the genotype of the mother, it will show that the maternal genotype is an important mediator. Specific Aim 2 will test the hypothesis that there are genetic differences in the dam that contribute to the efficacy of choline in mitigating ethanol's effects on cell death. B6 and BXD strains that show high levels of cell death following ethanol exposure will be tested with different doses of choline to find the lowest dose that is efficacious in ameliorating ethanol's effects. If the dose differs significantly acros the strains it will demonstrate that genotype is critical. In contrast, if choline's effects are equivalent across strains, it will suggest that genotype is unimportant and that choline should be equally beneficial in all FASD children and that once a relevant dose is found within the human population, it will likely be equally effective across a wide sector of the population. Specific Ai 3 will test the hypotheses that epigenetic changes induced by ethanol exposure differ based on genotype, as well as maternal environment, thus contributing to genetic variability in susceptibility to ethanol- induced neurotoxicity. Global methylation will be examined in the telencephalon using reduced representation bisulfite sequencing and bisulfite pyrosequencing and be compared to changes in gene expression. Information from this proposal will be important for determining 1) which genome to examine to identify children most at risk for specific types of ethanol-induced neuroteratological effects and 2) whether genotype needs to be considered in implementing choline as a therapeutic for ethanol-induced brain damage.
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0.939 |
2018 |
Goldowitz, Daniel Hamre, Kristin M |
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. |
Maternal Genotype, Choline Intervention, & Epigenetics in Fetal Alcohol Syndrome @ University of Tennessee Health Sci Ctr
The type and severity of ethanol-induced alterations following prenatal ethanol exposure is strongly impacted by a number of factors. A growing body of evidence suggests that sex is one of the important variables in ethanol's teratogenic effects. Further, recent data shows that acute ethanol exposure at the beginning of CNS development can have long-term sex-specific effects on behavior (Fish et al., Behav Brain Res. 338:173-184, 2018). This proposal will test the hypotheses that sex is an important variable in mediating ethanol's effects on the CNS just after neural tube closure, an early stage of CNS development. In the parent RO1, we are examining the role of genotype in ethanol's neuroteratogenesis, and sex and genotype are often interacting variables. These experiments will expand this by conducting an initial analysis of sex-by-genotype interactions by evaluating two different mouse strains. We will examine changes in DNA methylation in the telencephalon (primordial forebrain) of the developing neural tube because ethanol has been shown to alter DNA methylation and there is a wide literature on sex-specific differences in DNA methylation. Two groups of mice will be examined in each strain: ethanol-exposed given 5.9 g/kg ethanol in a binge model and controls given isocaloric maltose-dextrin. DNA methylation will be examined using an unbiased, whole-genome method, Reduced Representation Bisulfite Sequencing. Two strains will be examined and these were chosen because they exhibit differential sensitivity to prenatal alcohol exposure: 1 sensitive and 1 resistant to ethanol's effects. Data from these experiments fits the first of the stated goals of the NIH Strategic Plan for Women's Health Research from The Office of Research on Women's Health: increase sex differences research in basic science studies. These experiments will address several areas of interest under this goal including: 1) Evaluation of prenatal development, 2) Comparative study of cells, and specifically early?forming neural cells, from males and females, 3) Conduction of neuroscience research to understand vulnerability to ethanol-induced CNS damage, and 4) Evaluation of epigenetic modifications, specifically DNA methylation, using a whole-genome approach. Outcomes from this experiment will determine 1) Whether DNA methylation plays in a role in sex-specific neuroteratogenic effects following early ethanol exposure and 2) whether there is a sex-by-strain interaction suggesting that both variables are critically important in determining neuroteratogenicity. Additionally, the evaluation of sex differences in control embryos could provide information that would have a broader impact in our understanding of developmental conditions that have a sex ratio bias, such as autism. The ability to understand sex and sex-by-genotype differences in DNA methylation, particularly early in CNS development, will likely improve our understanding of how sex differences in brain function occur and potentially identify molecular pathways that mediate this difference.
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0.939 |