title: Forskolin and dopamine D1 receptor activation increase huntingtin's association with endosomes in immortalized neuronal cells of striatal origin.
abstract: Huntingtin is a cytoplasmic protein of unknown function that associates with vesicle membranes and microtubules. Its protein interactions suggest that huntingtin has a role in endocytosis and organelle transport. In this study we sought to identify factors that regulate the transport of huntingtin in striatal neurons, which are the cells most affected in Huntington's disease. In clonal striatal cells derived from fusions of neuroblastoma and embryonic striatal neurons, huntingtin localization is diffuse and slightly punctate in the cytoplasm. When these neurons were differentiated by treatment with forskolin, huntingtin redistributed to perinuclear regions, discrete puncta along plasma membranes, and branch points and terminal growth cones in neurites. Huntingtin staining overlapped with clathrin, a coat protein involved in endocytosis. Immunoblot analysis of subcellular membrane fractions separated by differential centrifugation confirmed that huntingtin immunoreactivity in differentiated neurons markedly increased in membrane fractions enriched with clathrin and with huntingtin-interacting protein 1. Dopamine treatment altered the subcellular localization of huntingtin and increased its expression in clathrin-enriched membrane fractions. The dopamine-induced changes were blocked by the D1 antagonist SCH 23390 and were absent in a clonal cell line lacking D1 receptors. Results suggest that the transport of huntingtin and its co-expression in clathrin and huntingtin-interacting protein 1-enriched membranes is influenced by activation of adenylyl cyclase and stimulation of dopamine D1 receptors.
authors.affiliation: Department of Neurology, Massachusetts General Hospital, Boston 02114, USA.
authors.forename: M
authors.initials: M
authors.lastname: Kim
authors.forename: J
authors.initials: J
authors.lastname: Velier
authors.forename: K
authors.initials: K
authors.lastname: Chase
authors.forename: G
authors.initials: G
authors.lastname: Laforet
authors.forename: M A
authors.initials: MA
authors.lastname: Kalchman
authors.forename: M R
authors.initials: MR
authors.lastname: Hayden
authors.forename: L
authors.initials: L
authors.lastname: Won
authors.forename: A
authors.initials: A
authors.lastname: Heller
authors.forename: N
authors.initials: N
authors.lastname: Aronin
authors.forename: M
authors.initials: M
authors.lastname: Difiglia
mesh_terms: D000818:Animals; D002461:Cell Line, Transformed; D002966:Clathrin / Q000378:metabolism; D005576:Colforsin / Q000494:pharmacology*; D003342:Corpus Striatum / Q000378:metabolism*; D004298:Dopamine / Q000494:pharmacology*; D004705:Endocytosis; D011992:Endosomes / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D000071058:Huntingtin Protein; D006816:Huntington Disease / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D009447:Neuroblastoma; D009474:Neurons / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D009687:Nuclear Proteins / Q000378:metabolism*; D017447:Receptors, Dopamine D1 / Q000187:drug effects / Q000502:physiology*; D014407:Tumor Cells, Cultured
chemical_list: D002966:Clathrin; C490758:Hap1 protein, mouse; C506812:Htt protein, mouse; D000071058:Huntingtin Protein; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D017447:Receptors, Dopamine D1; D005576:Colforsin; D004298:Dopamine