Angiotensin; receptors; protein purification; immunocytochemistry; cDNA cloning; behavior
Regulation of neuropeptide receptor gene expression and signal transduction in cultured neuronal cells and behaving animals
Radioligand binding techniques and receptor autoradiography; second messenger production; cell culture techniques; HPLC; receptor solubilization and immunoprecipitation; cDNA techniques; antisense technology; behavioral measurements
Our research focuses on the cellular and molecular mechanisms of angiotensin II (AngII) action in the brain. In particular, radioligand binding techniques, quantitative autoradiography, and immunocytochemistry are used to determine the localization of AngII receptors in the central nervous system, and biochemical techniques are then used to determine receptor coupling to ion channels and intracellular effectors such as adenylyl cyclase, nitric oxide syntase and phosphatidylinositol hydrolysis, in cultured neurons. Recent efforts include purification of neuronal AngII receptors, the development of receptor-directed polyclonal and monoclonal antibodies, as well as the cloning of the gene(s) which encode for these receptors. These molecular tools are then used to study the transcriptional and post-translational control of AngII receptor expression as it relates to the behavioral actions of this neuropeptide.
Fluharty, S.J., Reagan, L.P., He, P.F. and Yee, D.K. Heterogeneity of neuronal angiotensin type 2 (AT2) receptors Recent Advances in Angiotensin Receptors : 138-49, 1996.Immunohistochemical mapping of angiotensin Type 2 (AT2) receptors in rat brain Reagan, L.P., Flanagan-Cato, L.M., Yee, D.K., Ma, L.Y., Sakai, R.R. and Fluharty, S.J. Brain Research 662: 45-59, 1994.Sakai, R.R., Yang, X.D., He, P.F., Ma, L.Y., Guo, Y.F., Reilly, J.R., Moga, C.N. and Fluharty, S.J. Inhibition of receptor expression and behavioral actions of angiotensin II by intracerebroventricular AT1 receptor antisense oligonucleotides Journal of Neurochemistry 62: 2053-56, 1994.