{"id":{"repo_id":"vt","oai_identifier":"oai:vtechworks.lib.vt.edu:10919/77991"},"canonical_url":"https://search.dev.ndltd.org/etd/vt/oai:vtechworks.lib.vt.edu:10919/77991","repository":{"repo_id":"vt","name":"Virginia Tech","base_url":"https://vtechworks.lib.vt.edu/oai/request"},"display":{"title":"The Neurotoxicity of Insecticides to Striatal Dopaminergic Pathway","abstract":"Parkinson's disease (PD) is an age-related neurodegenerative disease, which is characterized by severe loss of dopaminergic neurons in the substantia nigra pars compacta (SNc) and consequent dopamine depletion in its projecting area. In this dissertation, I evaluated the neurotoxicity of several classes of insecticides/drugs/neurotoxins to the striatal dopaminergic pathway and their potential relationship to Parkinsonism in the C57BL/6 mouse model, using biochemical and molecular biology methods. In the first objective, I investigated the neurotoxicity in striatal dopaminergic pathways following co-application of permethrin (PM), chlorpyrifos (CPF) and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). The study was done because pyrethroid and organophosphorus compounds are widely used insecticides and they have been implicated in Gulf War Syndrome. We found that short-term, high-dose exposure to PM or CPF had no significant effects on the expression of dopamine transporter (DAT), tyrosine hydroxylase (TH), or α-synuclein protein in striatal nerve terminals, but the insecticides slightly enhanced the neurotoxicity of MPTP in C57BL/6 mice at 28 days post-treatment. This finding indicates a slowly developing neurotoxicity may occur after termination of high-dose exposure. Long-term, low-dose exposure to PM did not show significant neurotoxicity to striatal dopaminergic pathways when given alone, nor did this injection of PM enhance the neutotoxicity of MPTP in C57BL/6 mice. In addition, experiments with pure cis or trans isomers of permethrin showed that both cis and trans isomers contributed equally to the neurotoxicity of PM in the short-term high dose study. Previous studies demonstrated a deficiency in mitochondrial function in PD, and a high density of K⁺ATP channels are present in substantia nigra, which play an important role in the maintenance of the membrane potential under metabolic stress. Therefore, in the second objective, I investigated the effect of K⁺ATP channel blockage on the neurotoxicity of mitochondrial-directed neurotoxins to striatal dopaminergic pathways. I found that mitochondrial inhibitors are potent releasers of preloaded dopamine from striatal nerve terminals, with the most potent compounds active in the nanomolar range. Co-application of the K⁺ATP channel blocker glibenclamide selectively increased the dopamine-releasing effect by complex I inhibitors in vitro, and potentiated the neurotoxicity of MPTP (a complex I inhibitor) on DAT and TH expression, in vivo. Mechanistic studies demonstrated that mitochondrial inhibitor-induced dopamine release is Ca²⁺-dependent. In addition, the selectivity of glibenclamide is not correlated to ATP depletion, but associated with the generation of excessive reactive oxygen species at the site of complex I. In the third objective, I conducted comparative studies on the mode of action of rotenone-/reserpine-/tetrabenzaine (TBZ)-induced depletion, in vitro, as these three compounds share some similarities in their chemical structures. I found that rotenone, reserpine and TBZ selectively released preloaded dopamine and serotonin (5-HT), with the rank order as rotenone>reserpine>TBZ. Mechanistic studies demonstrated more than one mechanism was involved in both rotenone- and reserpine-induced neurotransmitter release. Ca²⁺-stimulated vesicular release and neurotransmitter transporter-mediated release are the common mechanisms involved in rotenone- and reserpine-induced dopamine release. Overall, the insecticides/drugs/neurotoxins tested in the above experiments all exhibited some effect on the nigrastrital dopaminergic pathway, either alone or by enhancing the toxicity of other chemicals in combination treatment.","abstract_html":"Parkinson's disease (PD) is an age-related neurodegenerative disease, which is characterized by severe loss of dopaminergic neurons in the substantia nigra pars compacta (SNc) and consequent dopamine depletion in its projecting area. In this dissertation, I evaluated the neurotoxicity of several classes of insecticides/drugs/neurotoxins to the striatal dopaminergic pathway and their potential relationship to Parkinsonism in the C57BL/6 mouse model, using biochemical and molecular biology methods. In the first objective, I investigated the neurotoxicity in striatal dopaminergic pathways following co-application of permethrin (PM), chlorpyrifos (CPF) and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). The study was done because pyrethroid and organophosphorus compounds are widely used insecticides and they have been implicated in Gulf War Syndrome. We found that short-term, high-dose exposure to PM or CPF had no significant effects on the expression of dopamine transporter (DAT), tyrosine hydroxylase (TH), or α-synuclein protein in striatal nerve terminals, but the insecticides slightly enhanced the neurotoxicity of MPTP in C57BL/6 mice at 28 days post-treatment. This finding indicates a slowly developing neurotoxicity may occur after termination of high-dose exposure. Long-term, low-dose exposure to PM did not show significant neurotoxicity to striatal dopaminergic pathways when given alone, nor did this injection of PM enhance the neutotoxicity of MPTP in C57BL/6 mice. In addition, experiments with pure cis or trans isomers of permethrin showed that both cis and trans isomers contributed equally to the neurotoxicity of PM in the short-term high dose study. Previous studies demonstrated a deficiency in mitochondrial function in PD, and a high density of K⁺ATP channels are present in substantia nigra, which play an important role in the maintenance of the membrane potential under metabolic stress. Therefore, in the second objective, I investigated the effect of K⁺ATP channel blockage on the neurotoxicity of mitochondrial-directed neurotoxins to striatal dopaminergic pathways. I found that mitochondrial inhibitors are potent releasers of preloaded dopamine from striatal nerve terminals, with the most potent compounds active in the nanomolar range. Co-application of the K⁺ATP channel blocker glibenclamide selectively increased the dopamine-releasing effect by complex I inhibitors in vitro, and potentiated the neurotoxicity of MPTP (a complex I inhibitor) on DAT and TH expression, in vivo. Mechanistic studies demonstrated that mitochondrial inhibitor-induced dopamine release is Ca²⁺-dependent. In addition, the selectivity of glibenclamide is not correlated to ATP depletion, but associated with the generation of excessive reactive oxygen species at the site of complex I. In the third objective, I conducted comparative studies on the mode of action of rotenone-/reserpine-/tetrabenzaine (TBZ)-induced depletion, in vitro, as these three compounds share some similarities in their chemical structures. I found that rotenone, reserpine and TBZ selectively released preloaded dopamine and serotonin (5-HT), with the rank order as rotenone>reserpine>TBZ. Mechanistic studies demonstrated more than one mechanism was involved in both rotenone- and reserpine-induced neurotransmitter release. Ca²⁺-stimulated vesicular release and neurotransmitter transporter-mediated release are the common mechanisms involved in rotenone- and reserpine-induced dopamine release. Overall, the insecticides/drugs/neurotoxins tested in the above experiments all exhibited some effect on the nigrastrital dopaminergic pathway, either alone or by enhancing the toxicity of other chemicals in combination treatment.","abstract_has_math":false,"creators":["Kou, Jinghong"],"institution":"Virginia Tech","degree_name":"Ph. D.","degree_level":"doctoral","degree_discipline":"Entomology","degree_department":"Entomology","school":null,"contributors":[],"advisors":[],"committee_chairs":["Bloomquist, Jeffrey R."],"committee_members":["Klein, Bradley G.","Paulson, Sally L.","Ehrich, Marion F.","Castagnoli, Kay P."],"year":2005,"date_issued":"2005-11-18","date_published":"2005-11-18","updated_at":"2026-07-22T22:18:44Z","subjects":["neurotoxicity testing","neurotoxicology","environmental neurotoxicant","neurodegeneration","neuroscience"],"languages":["en_US"],"rights":["In Copyright"],"rights_urls":["http://rightsstatements.org/vocab/InC/1.0/"],"identifier_entries":[{"key":"dc:identifier.other","label":"Dc Identifier Other","values":["etd-11302005-154325"],"render_values":[{"text":"etd-11302005-154325","href":null,"code":true}]}]},"links":{"outbound_url":"http://hdl.handle.net/10919/77991","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.committeechair","label":"Committee Chair","values":["Bloomquist, Jeffrey R."]},{"key":"dc:contributor.committeemember","label":"Committee Member","values":["Klein, Bradley G.","Paulson, Sally L.","Ehrich, Marion F.","Castagnoli, Kay P."]},{"key":"dc:contributor.department","label":"Department","values":["Entomology"]},{"key":"dc:creator","label":"Author","values":["Kou, Jinghong"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.accessioned","label":"Dc Date Accessioned","values":["2017-06-09T18:30:44Z"]},{"key":"dc:date.available","label":"Dc Date Available","values":["2017-06-09T18:30:44Z","2007-12-08"]},{"key":"dc:date.issued","label":"Date","values":["2005-11-18"]},{"key":"dc:publisher","label":"Institution","values":["Virginia Tech"]},{"key":"dc:type","label":"Dc Type","values":["Dissertation"]},{"key":"dc:type.dcmitype","label":"Dc Type Dcmitype","values":["Text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Entomology"]},{"key":"thesis:degree_level","label":"Degree Level","values":["doctoral"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Ph. 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In this dissertation, I evaluated the neurotoxicity of several classes of insecticides/drugs/neurotoxins to the striatal dopaminergic pathway and their potential relationship to Parkinsonism in the C57BL/6 mouse model, using biochemical and molecular biology methods. In the first objective, I investigated the neurotoxicity in striatal dopaminergic pathways following co-application of permethrin (PM), chlorpyrifos (CPF) and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). The study was done because pyrethroid and organophosphorus compounds are widely used insecticides and they have been implicated in Gulf War Syndrome. We found that short-term, high-dose exposure to PM or CPF had no significant effects on the expression of dopamine transporter (DAT), tyrosine hydroxylase (TH), or α-synuclein protein in striatal nerve terminals, but the insecticides slightly enhanced the neurotoxicity of MPTP in C57BL/6 mice at 28 days post-treatment. This finding indicates a slowly developing neurotoxicity may occur after termination of high-dose exposure. Long-term, low-dose exposure to PM did not show significant neurotoxicity to striatal dopaminergic pathways when given alone, nor did this injection of PM enhance the neutotoxicity of MPTP in C57BL/6 mice. In addition, experiments with pure cis or trans isomers of permethrin showed that both cis and trans isomers contributed equally to the neurotoxicity of PM in the short-term high dose study. Previous studies demonstrated a deficiency in mitochondrial function in PD, and a high density of K⁺ATP channels are present in substantia nigra, which play an important role in the maintenance of the membrane potential under metabolic stress. Therefore, in the second objective, I investigated the effect of K⁺ATP channel blockage on the neurotoxicity of mitochondrial-directed neurotoxins to striatal dopaminergic pathways. I found that mitochondrial inhibitors are potent releasers of preloaded dopamine from striatal nerve terminals, with the most potent compounds active in the nanomolar range. Co-application of the K⁺ATP channel blocker glibenclamide selectively increased the dopamine-releasing effect by complex I inhibitors in vitro, and potentiated the neurotoxicity of MPTP (a complex I inhibitor) on DAT and TH expression, in vivo. Mechanistic studies demonstrated that mitochondrial inhibitor-induced dopamine release is Ca²⁺-dependent. In addition, the selectivity of glibenclamide is not correlated to ATP depletion, but associated with the generation of excessive reactive oxygen species at the site of complex I. In the third objective, I conducted comparative studies on the mode of action of rotenone-/reserpine-/tetrabenzaine (TBZ)-induced depletion, in vitro, as these three compounds share some similarities in their chemical structures. I found that rotenone, reserpine and TBZ selectively released preloaded dopamine and serotonin (5-HT), with the rank order as rotenone>reserpine>TBZ. Mechanistic studies demonstrated more than one mechanism was involved in both rotenone- and reserpine-induced neurotransmitter release. Ca²⁺-stimulated vesicular release and neurotransmitter transporter-mediated release are the common mechanisms involved in rotenone- and reserpine-induced dopamine release. Overall, the insecticides/drugs/neurotoxins tested in the above experiments all exhibited some effect on the nigrastrital dopaminergic pathway, either alone or by enhancing the toxicity of other chemicals in combination treatment."]},{"key":"dc:description.degree","label":"Dc Description Degree","values":["Ph. D."]},{"key":"dc:title","label":"Title","values":["The Neurotoxicity of Insecticides to Striatal Dopaminergic Pathway"]}]}],"canonical_facts":{"dc:contributor.committeechair":["Bloomquist, Jeffrey R."],"dc:contributor.committeemember":["Klein, Bradley G.","Paulson, Sally L.","Ehrich, Marion F.","Castagnoli, Kay P."],"dc:contributor.department":["Entomology"],"dc:creator":["Kou, Jinghong"],"dc:date.accessioned":["2017-06-09T18:30:44Z"],"dc:date.available":["2017-06-09T18:30:44Z","2007-12-08"],"dc:date.issued":["2005-11-18"],"dc:description.abstract":["Parkinson's disease (PD) is an age-related neurodegenerative disease, which is characterized by severe loss of dopaminergic neurons in the substantia nigra pars compacta (SNc) and consequent dopamine depletion in its projecting area. In this dissertation, I evaluated the neurotoxicity of several classes of insecticides/drugs/neurotoxins to the striatal dopaminergic pathway and their potential relationship to Parkinsonism in the C57BL/6 mouse model, using biochemical and molecular biology methods. In the first objective, I investigated the neurotoxicity in striatal dopaminergic pathways following co-application of permethrin (PM), chlorpyrifos (CPF) and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). The study was done because pyrethroid and organophosphorus compounds are widely used insecticides and they have been implicated in Gulf War Syndrome. We found that short-term, high-dose exposure to PM or CPF had no significant effects on the expression of dopamine transporter (DAT), tyrosine hydroxylase (TH), or α-synuclein protein in striatal nerve terminals, but the insecticides slightly enhanced the neurotoxicity of MPTP in C57BL/6 mice at 28 days post-treatment. This finding indicates a slowly developing neurotoxicity may occur after termination of high-dose exposure. Long-term, low-dose exposure to PM did not show significant neurotoxicity to striatal dopaminergic pathways when given alone, nor did this injection of PM enhance the neutotoxicity of MPTP in C57BL/6 mice. In addition, experiments with pure cis or trans isomers of permethrin showed that both cis and trans isomers contributed equally to the neurotoxicity of PM in the short-term high dose study. Previous studies demonstrated a deficiency in mitochondrial function in PD, and a high density of K⁺ATP channels are present in substantia nigra, which play an important role in the maintenance of the membrane potential under metabolic stress. Therefore, in the second objective, I investigated the effect of K⁺ATP channel blockage on the neurotoxicity of mitochondrial-directed neurotoxins to striatal dopaminergic pathways. I found that mitochondrial inhibitors are potent releasers of preloaded dopamine from striatal nerve terminals, with the most potent compounds active in the nanomolar range. Co-application of the K⁺ATP channel blocker glibenclamide selectively increased the dopamine-releasing effect by complex I inhibitors in vitro, and potentiated the neurotoxicity of MPTP (a complex I inhibitor) on DAT and TH expression, in vivo. Mechanistic studies demonstrated that mitochondrial inhibitor-induced dopamine release is Ca²⁺-dependent. In addition, the selectivity of glibenclamide is not correlated to ATP depletion, but associated with the generation of excessive reactive oxygen species at the site of complex I. In the third objective, I conducted comparative studies on the mode of action of rotenone-/reserpine-/tetrabenzaine (TBZ)-induced depletion, in vitro, as these three compounds share some similarities in their chemical structures. I found that rotenone, reserpine and TBZ selectively released preloaded dopamine and serotonin (5-HT), with the rank order as rotenone>reserpine>TBZ. Mechanistic studies demonstrated more than one mechanism was involved in both rotenone- and reserpine-induced neurotransmitter release. Ca²⁺-stimulated vesicular release and neurotransmitter transporter-mediated release are the common mechanisms involved in rotenone- and reserpine-induced dopamine release. Overall, the insecticides/drugs/neurotoxins tested in the above experiments all exhibited some effect on the nigrastrital dopaminergic pathway, either alone or by enhancing the toxicity of other chemicals in combination treatment."],"dc:description.degree":["Ph. D."],"dc:identifier.other":["etd-11302005-154325"],"dc:identifier.uri":["http://hdl.handle.net/10919/77991"],"dc:language.iso":["en_US"],"dc:publisher":["Virginia Tech"],"dc:rights":["In Copyright"],"dc:rights.uri":["http://rightsstatements.org/vocab/InC/1.0/"],"dc:subject":["neurotoxicity testing","neurotoxicology","environmental neurotoxicant","neurodegeneration","neuroscience"],"dc:title":["The Neurotoxicity of Insecticides to Striatal Dopaminergic Pathway"],"dc:type":["Dissertation"],"dc:type.dcmitype":["Text"],"thesis:degree_discipline":["Entomology"],"thesis:degree_level":["doctoral"],"thesis:degree_name":["Ph. D."],"thesis:institution_name":["Virginia Polytechnic Institute and State University"]},"updated_at":"2026-07-22T22:18:44Z"}