1985 — 1986 |
Bohn, Martha C |
K04Activity Code Description: Undocumented code - click on the grant title for more information. |
Development of the Adrenergic Phenotype in Brain @ State University New York Stony Brook
The proposed research is directed toward the long term goal of defining developmental factors which are crucial in differentiation and specification of neurons. Techniques previously applied to describe neurotransmitter phenotypic expression and development in the peripheral nervous system will be applied to the brain. Specifically, the embryonic and postnatal development of the adrenergic C1-C3 cell groups of the rat medulla oblongata will be studied to: 1) document the initial appearance of a range of adrenergic phenotypic characters, 2) define the temporal relationship between "neuronal birthdays" and the initial expression of adrenergic phenotypic characters, 3) compare central and peripheral adrenergic neurons witb respect to phenotypic expression, development and regulation by glucocorticoids, 4) elucidate possible transplacental effects of maternal glucocorticoids on adrenergic development in fetal brain, 5) investigate peptidergic expression in central adrenergic neurons, and 6) study the effects of target structures on adrenergic development. An interdisciplinary approach involving immunocytochemistry, biochemistry, autoradiography, pharmacology and cell culture will be taken. These studies promise to elucidate loci where changes in the milieu of the developing neuron may lead to abnormal neuronal development and ultimately to therapeutic approaches for the correction of neuronal deficits.
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0.942 |
1985 — 1988 |
Bohn, Martha C |
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. |
Embryonic Development of Epinergic Neurons in Brain @ State University New York Stony Brook
Epinergic neurons express specific enzymes required for the synthesis of epinephrine. The proposed studies will investigate the embryonic and postnatal differentiation of epinergic neurons in the rat brainstem and epinergic fibers in spinal cord and hypothalamus. In addition, possible hormonal influences on the development of epinergic phenotypic markers will be studied in vivo and in vitro. Immunocytochemistry, autoradiography, high-performance liquid chromatography, radioenzymatic assay and tissue culture will be combined to provide morphological and biochemical descriptions of epinergic differentiation. Specifically, the proposed studies seek to 1) define the temporal relationships between neurogenesis and epinergic phenotypic expression in neural tube and neural crest derivatives, 2) investigate the phenomenon of transient neurotransmitter expression in the brain, 3) describe the development of epinergic fibers in spinal cord and hypothalamus, 4) compare central to peripheral epinergic cells with respect to possible regulation by glucocorticoids, 5) elucidate possible transplacental effects of glucocorticoids on epinergic development in the fetal brain and spinal cord, 6) investigate expression of peptidergic phenotypes in epinergic neurons, and 7) study factors which influence the in vitro development of epinergic neurons in long-term explant cultures of embryonic medulla oblongata. These studies address fundamental issues in developmental biology and neuroscience concerning the processes involved in cell determination, specificity and differentiation. Elucidation of factors involved in neurotransmitter expression and development in brain and the possible role of hormones as extracellular influences, may, in the future, lead to an understanding of loci where changes in the milieu of the developing neuron produces aberrant development. In turn, this may contribute to an understanding of birth defects in the human and ultimately to therapeutic manipulation of processes involved in neuronal differentiation.
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0.975 |
1989 |
Bohn, Martha C |
S10Activity Code Description: To make available to institutions with a high concentration of NIH extramural research awards, research instruments which will be used on a shared basis. |
Protein Database Workstation @ University of Rochester
A shared resource for the computerized analysis of proteins separated by two-dimensional gel electrophoresis and the compilation of information to build protein databases is requested. This facility would be comprised of commercially available software marketed by Protein Databases, Inc. for the scanning and quantitation of two-dimensional gels and a Sun computer system with graphic capabilities. This resource represents instrumentation and software which is on the forefront of protein analysis and would be one of less than a dozen such facilities in the world. Users will be able to scan and digitize autoradiograms from gels run in their laboratory or run commercially, as well as silver stained gels. The software will allow matching of the patterns on the digitized gel images, quantitation of thousands or proteins, the building of protein databases, and the use of public databases. users will have access to two terminals and will output their data by several means, including numerical quantitation, graphs representing experimental changes in each protein spot and large spot maps depicting a synthetic gel image of proteins located by molecular weight, isoelectric point and data base number. The long-term goals to be addressed using this facility are to identify proteins involved in cell specificity, cell differentiation and cell regulation. In addition, the facility will be used to communicate information on proteins from experiment to experiment, as well as from laboratory to laboratory around the world through the use of public databases. The collaborative user group requesting this instrumentation will rely heavily on this resource to identify proteins involved in 1) neuronal and glial cell differentiation, 2) glucocorticoid and growth factor action, 3) cell transformation and oncogenesis, 4) cell responses to neurotoxins, 5) hematopoiesis, 6) membrane transport systems, and 7) Alzheimer's disease and aging. The unique application of applying this commercially available software to the analysis of ultrahigh-resolution giant 2D gels is also proposed. Since the proposed facility has the capacity to quantify and store information on thousands of proteins, it promises to significantly enhance progress toward the identification of proteins involved in disease.
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0.975 |
1993 |
Bohn, Martha C |
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. |
Glucocorticoids and Brain Development @ University of Rochester
The long term goal of these studies is to define the molecular mechanisms involved in adrenal glucocorticoid regulation of brain development. The studies proposed here will begin by studying the expression and development of glucocorticoid- (GR) and mineralocorticoid- (MR) receptors and identifying proteins that are regulated by glucocorticoids in hippocampal neurons and glial cells. In the future, these studies promise to lead to the cloning and sequencing of those genes and gene products that are regulated by glucocorticoids at the transcriptional level in specific classes of neurons and glia. Our previous work on glucocorticoid effects on neural development and glucocorticoid receptor expression in the brain places us in a position to address these issues. An interdisciplinary approach using primary cell culture, immunocytochemistry, in situ hybridization, and quantitative analysis of proteins separated by ultra-high resolution giant 2-D gel electrophoresis will be applied. Specifically, the proposed studies seek to a) describe the initial expression and ontogeny of GRs and MRs and their corresponding mRNAs in the fetal and postnatal rat brain, b) determine which classes of glia express GRs or MRs, c) study the possible co-expression of GRs and MRs within the same cell, d) identify proteins regulated by glucocorticoids in astrocytes, oligodendrocytes, hippocampal neurons and HT4 cells, an immortalized clonal neuronal cell line derived from hippocampus e) investigate whether glucocorticoid effects on the metabolism of specific proteins are mediated by the MR or the GR, or both, and f) determine whether glucocorticoid regulation of specific proteins occurs at the level of mRNA. More generally, these studies address current issues in developmental neurobiology concerning the role of extracellular factors in cell differentiation and regulation. These studies will increase our understanding of mechanisms through which hormones derived from the periphery influence differentiation of neurons and glia and cellular regulation in the adult brain. Moreover, these studies promise to elucidate loci where changes in the hormonal milieu of cells in the brain, elicited by disease, steroid therapy or aging, may lead to behavioral abnormalities. Such knowledge may contribute to our understanding of birth defects, senility and depression in the human and ultimately lead to therapeutic approaches to treat these disorders.
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0.975 |
2002 |
Bohn, Martha C |
U54Activity 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 differ from program project 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, with funding component staff helping to identify appropriate priority needs. |
Tet Regulated Vectors For Parkinson's Disease @ University of Rochester
The long-term goal of this project is to develop viral vectors that can be used as gene therapy vehicles to treat Parkinson's disease (PD), as well as other chronic neurodegenerative diseases. Such vectors require the incorporation of a promoter element that permits effective and safe regulation of the therapeutic gene through peripheral drug administration. The proposed experiments will focus on tetracycline(tet)-regulated promoter systems in the context of recombinant adeno-associated viral vectors (AAV) and tentiviral vectors which are based on the human immunodeficiency virus (HIV). A set of AAV and HIV viral vectors will be made that incorporate cellular marker and therapeutic genes driven by tet-regulated promoters that can be turned-on or turned-off by administration of the tet analog doxycycline (Dox). Cellular marker genes including humanized green fluorescent protein (hrGFP) and an non-immunogenic gene, rat alkaline phosphatase (rAP), will be used to assess the efficiency and suitability of the vector designs. Quantitative assays including flow cytometry, real-time, quantitative reverse transcriptase-polymerase chain reaction (QRT-PCR), ELISA and computerized morphometry will be applied to assess and compare vectors in cell culture and in the rat nigrostriatal system. In vivo studies will be clone in both normal intact rat brain and in the 6-OHDA progressively lesioned rat model of PD to determine whether damage related to PD will affect the efficiency of vectors containing regulated promoters. These studies will also determine to what extent the therapeutic effects of glial cell line-derived neurotrophic factor (GDNF) gene therapy in this rat model of PD are reversible. Effects of GDNF gene delivery using tet-regulated vectors on dopamine neurons will be evaluated using quantitative morphometric, molecular and behavioral evaluations. All studies will involve assessment of host immune reactions and chromosomal effects of the vectors in collaboration with the Lowenstein and Federoff labs in this consortium. The successful generation of a viral vector that fulfills the requirements of tight regulation, long-term expression and regulatability with minimal host immune responses in the rat CNS will be advanced to non-human primate preclinical trials for PD.
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0.975 |