The long term objectives of this research plan are to develop immunological probes to specific synaptic glycoproteins in the chicken retina in order to determine the functional significance of these molecules with respect to the formation of and maintenance of synapses. These objectives are of special relevance to studies concerned with the normal development of neural pathways and also studies concerned with neural regeneration following recovery from various neurological disorders. Specifically, the aims of the project are to determine the molecular weights and lectin specificities of individual glycoproteins associated with synaptic plasma membranes and synaptic junctions isolated from the chick retina. Further aspects of this application include the description of cytoskeletal proteins involved in the organization of particular surface glycoproteins in the synapse and the isolation of glycoprotein-binding molecules or endogenous lectins that are associated with specific glycoproteins. Essentially, the methods involve a characterization of lectin-binding affinities of individual glycoproteins within the synaptic array of proteins observed in polyacrylamide gel-nitrocellulose paper blots which we have shown to be a suitable substrate for this purpose. This will provide us with information concerning the presence of and types of sugar residues that are accessible to lectin probes in individual synaptic proteins. Secondly, glycoproteins, endogenous lectins, and glycoprotein associated cytoskeletal proteins will be identified and characterized through use of affinity column chromatography techniques. Affinity isolated glycoproteins and endogenous lectins will be used to produce rabbit antisera against these purified molecules. The antisera will be used first to localize through electron microscope immunocytochemistry these specific molecules in tissue sections of the retina. This localization can be matched to specific molecules identified with lectins and antisera on gel-blots of the isolated synaptic fractions. In the future these immunological probes can be used to study transport and turnover of particular synaptic molecules. They will also be essential for the development of assays of functional relationships between specific molecules and synapse formation during development and regeneration.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY002708-08
Application #
3257080
Study Section
Visual Sciences A Study Section (VISA)
Project Start
1978-09-29
Project End
1987-11-30
Budget Start
1985-12-01
Budget End
1986-11-30
Support Year
8
Fiscal Year
1986
Total Cost
Indirect Cost
Name
University of Tennessee Health Science Center
Department
Type
Schools of Medicine
DUNS #
941884009
City
Memphis
State
TN
Country
United States
Zip Code
38163
Liu, N; Cooper, N G (1996) The Ca2+/calmodulin-dependent protein kinase II-associated protein complex isolated from chicken retina. J Mol Neurosci 7:1-12
Pozdnyakov, N; Yoshida, A; Cooper, N G et al. (1995) A novel calcium-dependent activator of retinal rod outer segment membrane guanylate cyclase. Biochemistry 34:14279-83
Cooper, N; Liu, L; Yoshida, A et al. (1995) The bovine rod outer segment guanylate cyclase, ROS-GC, is present in both outer segment and synaptic layers of the retina. J Mol Neurosci 6:211-22
Cooper, N G; Wei, X; Liu, N (1995) Onset of expression of the alpha subunit of Ca2+/calmodulin-dependent protein kinase II and a novel related protein in the developing retina. J Mol Neurosci 6:75-89
Liu, N; Cooper, N G (1994) Purification and characterization of the Ca2+/calmodulin-dependent protein kinase II from chicken forebrain. J Mol Neurosci 5:193-206
Tien, L F; McLaughlin, B J; Cooper, N G (1991) Glycoproteins in the retinal pigment epithelium of normal and dystrophic rats. Invest Ophthalmol Vis Sci 32:319-26
Chang, H T; Kuo, H; Whittaker, J A et al. (1990) Light and electron microscopic analysis of projection neurons retrogradely labeled with Fluoro-Gold: notes on the application of antibodies to Fluoro-Gold. J Neurosci Methods 35:31-7
Waters, R S; McCandlish, C A; Cooper, N G (1990) Early development of SI cortical barrel subfield representation of forelimb in normal and deafferented neonatal rat as delineated by peroxidase conjugated lectin, peanut agglutinin (PNA). Exp Brain Res 81:234-40
Steindler, D A; Cooper, N G; Faissner, A et al. (1989) Boundaries defined by adhesion molecules during development of the cerebral cortex: the J1/tenascin glycoprotein in the mouse somatosensory cortical barrel field. Dev Biol 131:243-60
Cooper, N G; Steindler, D A (1989) Critical period-dependent alterations of the transient body image in the rodent cerebral cortex. Brain Res 489:167-76

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