A greater understanding of the intracellular signal transduction mechanisms which underlie dopaminergic neurotransmission in the basal ganglia can lead to novel therapeutic targets for the treatment of schizophrenia, and degenerative neurological disorders such as Parkinson's disease and Huntington's disease. A multi-disciplinary approach will be undertaken in the current proposal to investigate the phosphorylation-dependent functional regulation of key molecules in these signaling pathways. Studies will be performed at several levels of organizational complexity, ranging from in vitro biochemical studies with purified molecules to the characterization of phenotypes resulting from targeted deletion of key signal transduction molecules in whole animals. The overall goal of this project is to provide a Scientific Core facility for use by all the members of the Program Project. The responsibilities of the Core will include the production and supply of key reagents and the performance of routine, yet critical tasks that will be required to accomplish the studies described in the other sections of this Program Project. The centralization of routine tasks will facilitate an efficient, cost-effective means to ensure an adequate supply of essential materials and reagents to all the Projects. The Core will maintain a tissue culture facility to provide a continuous supply of cultured cells, including primary neuronal cultures, reaggregated cultures and immortalized cell lines. It will also maintain stocks of purified protein kinases, protein phosphatases, antibodies, and substrate proteins. Standard purification protocols will be used to obtain these enzymes from native sources, and recombinant technologies will also be employed to produce specific proteins in quantities sufficient to carry out detailed enzymological and structural studies.. The Core will be responsible for the expansion and maintenance of the colonies of """"""""knockout"""""""" mice that are deficient in DARPP-32, inhibitor-1, protein phosphatase-1 alpha1 and gamma isoforms, and protein phosphatase- 2B, and transgenic animals harboring phosphorylation site mutations in DARPP-32.. Animal husbandry will include the setting up of appropriate matings and weaning of animals. The Core will also be responsible for the determination of the genotype of the offspring, using PCR and Southern blot strategies. The Core will be responsible for the production of phosphorylation state- specific antibodies to specific sites on DARPP-32, inhibitor-1, protein phosphatase-1, Na+,K+-ATPase alpha subunit isoforms and Na+ and Ca2+ channels. The ability to detect and quantitate changes in the state of phosphorylation of these key substrate proteins will be of great utility in the studies proposed to elucidate their functional significance.

Agency
National Institute of Health (NIH)
Institute
National Institute of Mental Health (NIMH)
Type
Research Program Projects (P01)
Project #
5P01MH040899-14
Application #
6273398
Study Section
Project Start
1998-07-01
Project End
1999-06-30
Budget Start
1997-10-01
Budget End
1998-09-30
Support Year
14
Fiscal Year
1998
Total Cost
Indirect Cost
Name
Rockefeller University
Department
Type
DUNS #
071037113
City
New York
State
NY
Country
United States
Zip Code
10065
Plattner, Florian; Hayashi, Kanehiro; Hernández, Adan et al. (2015) The role of ventral striatal cAMP signaling in stress-induced behaviors. Nat Neurosci 18:1094-100
Kimura, Toru; Han, Wonsun; Pagel, Philipp et al. (2011) Protein phosphatase 2A interacts with the Na,K-ATPase and modulates its trafficking by inhibition of its association with arrestin. PLoS One 6:e29269
Zhou, Mingming; Rebholz, Heike; Brocia, Christine et al. (2010) Forebrain overexpression of CK1delta leads to down-regulation of dopamine receptors and altered locomotor activity reminiscent of ADHD. Proc Natl Acad Sci U S A 107:4401-6
Bertran-Gonzalez, Jesus; HÃ¥kansson, Kerstin; Borgkvist, Anders et al. (2009) Histone H3 phosphorylation is under the opposite tonic control of dopamine D2 and adenosine A2A receptors in striatopallidal neurons. Neuropsychopharmacology 34:1710-20
Kuroiwa, Mahomi; Bateup, Helen S; Shuto, Takahide et al. (2008) Regulation of DARPP-32 phosphorylation by three distinct dopamine D1-like receptor signaling pathways in the neostriatum. J Neurochem 107:1014-26
Nishi, Akinori; Kuroiwa, Mahomi; Miller, Diane B et al. (2008) Distinct roles of PDE4 and PDE10A in the regulation of cAMP/PKA signaling in the striatum. J Neurosci 28:10460-71
Barbano, Paolo E; Spivak, Marina; Flajolet, Marc et al. (2007) A mathematical tool for exploring the dynamics of biological networks. Proc Natl Acad Sci U S A 104:19169-74
Borgkvist, Anders; Usiello, Alessandro; Greengard, Paul et al. (2007) Activation of the cAMP/PKA/DARPP-32 signaling pathway is required for morphine psychomotor stimulation but not for morphine reward. Neuropsychopharmacology 32:1995-2003
Bullock, S Andrew; Platholi, Jimcy; Gjyrezi, Ada et al. (2007) Differential regulation of protein phosphatase-1(I) by neurabin. Biochem Biophys Res Commun 358:140-4
Svenningsson, Per; Bateup, Helen; Qi, Hongshi et al. (2007) Involvement of AMPA receptor phosphorylation in antidepressant actions with special reference to tianeptine. Eur J Neurosci 26:3509-17

Showing the most recent 10 out of 219 publications