Effects of basic fibroblast growth factor (bFGF) on the development of midbrain dopamine neurons in vitro.
Item
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Title
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Effects of basic fibroblast growth factor (bFGF) on the development of midbrain dopamine neurons in vitro.
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Identifier
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AAI9605575
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identifier
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9605575
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Creator
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Bouvier, Margaret Marie.
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Contributor
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Adviser: Catherine Mytilineou
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Date
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1995
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Language
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English
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Publisher
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City University of New York.
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Subject
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Biology, Neuroscience | Health Sciences, Medicine and Surgery
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Abstract
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Parkinson's disease is characterized by a loss of dopamine neurons in the substantia nigra leading to decreased dopamine in its target structure, the striatum. Drug therapy is commonly used to treat the symptoms of this disease, but it is complicated by side effects and limited efficacy. More recently, transplantation of substantia nigra primordia to the striatum has yielded encouraging preliminary results. This therapy would be improved by developing ways to expand in vitro the number of dopamine precursors, in order to generate large enough quantities of appropriate-aged, dopamine enriched tissue which would be available as needed for surgery.;The goal of this research was to explore the possibility of various factors to manipulate the proliferation and differentiation of dopamine precursors. Cultures were prepared from embryonic day 12 (E12) rat ventral mesencephalon, coinciding with the beginning of the birth of dopamine neurons of the substantia nigra. At low plating density in serum-free medium, dopamine precursors divide for approximately one day in vitro. We report here that basic fibroblast growth factor (bFGF) expands the period of dopamine precursor division at least until day 8 in culture with the majority dividing on days 4-5, well beyond their normal division. This increase in cell division was accompanied by a delay in differentiation as compared to untreated control cultures. Upon differentiation, the dopamine neurons in the bFGF-treated cultures yielded high-affinity dopamine uptake values that were twenty times maximal control values.;Culturing E12 cells on a non-adhesive substrate generated a suspension of spheres, which is more suitable for transplantation. bFGF stimulated cell proliferation in these spheres; control cultures demonstrated limited division and survival. TH+ cells were not detected in spheres cultured for 5 days in vitro (DIV), but were found in spheres at DIV 7, 9, and 12. When DIV 12 spheres were attached to an adhesive substrate and further cultured, dopamine neurons migrated out of large spheres, forming extensive fiber networks. These large dopamine-containing spheres constituted approximately 25% of the spheres in a typical culture dish, and exhibited uptake values approximately 6 times higher than controls.
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Type
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dissertation
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Source
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PQT Legacy CUNY.xlsx
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degree
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Ph.D.
