Studies will be undertaken to clone prospective alpha2-adrenergic receptor subtypes and to understand the molecular structure of alpha2-adrenergic receptors. At present the alpha2-adrenergic receptors can be classified into the alpha2-C10, alpha2-C4 and alpha2-C2 subtypes on the basis of molecular biological studies. On the basis of pharmacological studies, the alpha2-adrenergic receptors can be classified clearly into the alpha2A and alpha2B subtypes; additionally, there is suggestive evidence for two more subtypes. The alpha2-C10 and alpha2A subtypes are equivalent and recent data indicates that the alpha2-C2 and alpha2B subtypes are the same. The relationship of the alpha2-C4 to the pharmacologically defined subtypes is less clear, although, it may be equivalent to the alpha2B as characterized in brain tissue. Preliminary results clearly support the possibility of a fourth alpha2-adrenergic receptor subtype. The known DNA sequences of the alpha2-C10, alpha2-C4 and alpha2-C2 subtypes will be used to design oligonucleotide probes for use in the polymerase chain reaction (PCR). This powerful technique, which can be used to clone related genes, will be used in conjunction with traditional methods to identify new alpha2- receptor subtypes. Mutagenesis studies will be done to understand and define the ligand binding site of present and future members of alpha2-adrenergic receptor family. These studies will involve site-directed mutagenesis and PCR-based approaches. Recombinant DNA techniques will also be used to study palmitoylation of the alpha2-adrenergic receptor subtypes and to design receptor-fusion proteins that can be expressed in prokaryotic cells. These studies will help characterize important biochemical features of the alpha2-adrenergic receptor subtypes. The information that will be obtained by the proposed studies will help us understand the pharmacological characteristics of alpha2-adrenergic receptor subtypes. Drugs acting via alpha2-adrenergic receptors are presently used as sedatives and to treat hypertension. Additionally, alpha2-adrenergic agents have potential in the treatment of drug abuse, diabetes, memory loss and weight control. Some of these conditions represent serious public health problems that are difficult to treat and that may get worse as our nation deals with drug abuse and an aging population. The work being proposed here will provide a basis from which therapeutic solutions to these problems may be found.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
First Independent Research Support & Transition (FIRST) Awards (R29)
Project #
5R29EY009355-03
Application #
3465897
Study Section
Pharmacology A Study Section (PHRA)
Project Start
1991-04-01
Project End
1996-03-31
Budget Start
1993-04-01
Budget End
1994-03-31
Support Year
3
Fiscal Year
1993
Total Cost
Indirect Cost
Name
University of Arizona
Department
Type
Schools of Pharmacy
DUNS #
City
Tucson
State
AZ
Country
United States
Zip Code
85721
Porter, Amy C; Svensson, Samuel P S; Stamer, W Daniel et al. (2003) Alpha-2 adrenergic receptors stimulate actin organization in developing fetal rat cardiac myocytes. Life Sci 72:1455-66
Huang, Yi; Stamer, W Daniel; Anthony, Todd L et al. (2002) Expression of alpha(2)-adrenergic receptor subtypes in prenatal rat spinal cord. Brain Res Dev Brain Res 133:93-104
Anthony, T L; Pierce, K L; Regan, J W (2000) Characterization of ep prostanoid receptor subtypes in primary cultures of bovine ciliary epithelial cells by immunofluorescent microscopy and functional studies. Curr Eye Res 20:394-404
Fraeyman, N; Vanscheeuwijck, P; Wang, J M et al. (1999) Changes in the expression of alpha(2)-adrenergic receptor subtypes during maturation of neuronal cells from fetal pig superior cervical ganglia. Brain Res Dev Brain Res 116:127-32
Richman, J G; Regan, J W (1998) Alpha 2-adrenergic receptors increase cell migration and decrease F-actin labeling in rat aortic smooth muscle cells. Am J Physiol 274:C654-62
Anthony, T L; Pierce, K L; Stamer, W D et al. (1998) Prostaglandin F2 alpha receptors in the human trabecular meshwork. Invest Ophthalmol Vis Sci 39:315-21
Svensson, S P; Porter, A C; Regan, J W (1997) Cloning and functional expression of guinea pig atrial alpha 2-adrenergic receptor subtypes. Ann N Y Acad Sci 812:171-3
Pierce, K L; Bailey, T J; Hoyer, P B et al. (1997) Cloning of a carboxyl-terminal isoform of the prostanoid FP receptor. J Biol Chem 272:883-7
Stamer, W D; Snyder, R W; Regan, J W (1996) Characterization of the transmembrane orientation of aquaporin-1 using antibodies to recombinant fusion proteins. Biochemistry 35:16313-8
Stamer, W D; Huang, Y; Seftor, R E et al. (1996) Cultured human trabecular meshwork cells express functional alpha 2A adrenergic receptors. Invest Ophthalmol Vis Sci 37:2426-33

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