Glycoproteins are thought to play an important role during the formation of synapses. The goal of this research proposal is to examine the hypothesis that glycoproteins are essential substrates for the formation and maintenance of synapses in the retina. The implication that glycoproteins are also involved in mechanisms underlying synaptic specificity will also be evaluated by searching for antibodies that bind to the different types of synapses present in the retina. The general approach to this problem will be to produce mouse and/or rabbit antibodies to specific chick retinal synaptic glycoproteins in order that they may be localized. In order to maximize the chances of obtaining antibodies to the antigens of interest such as: a) synapse-specific glycoproteins, b) retinal-synapse, specific glycoproteins and c) retinal cell class-specific, synaptic glycoproteins, several subfractions of the retina will be isolated and used a immunogens. The subfractions will include: 1) retinal synaptic plasma membranes (SPM), 2) retinal synaptic junction complexes (SJC), 3) lectin affinity isolated glycoproteins and 4) cone-specific subcellular fractions, all of which can be reliably isolated and characterized (progress report). Antibodies will be assayed for their abilities to bind to tissue sections of the retina and/or to Western blots of the subfractions. A combined lectin and immunoprobe analysis of Western blots of these fractions will allow identification of antibodies to specific, characterized glycoproteins. This screening will allow us to identify antibodies that will be useful for analysis in tissue localization studies. An electron microscope, immunocytochemical analysis of developing retinal synapses in embryonic and hatched chicks will be performed to determine the time of appearance and mode of insertion of specific glycoproteins into synapses. The data derived from this project will help elucidate mechanisms underlying synaptogenesis and will be most useful for characterizing development of patterns of specific classes of synapses within the retina. Evidence of accomplishment in each of the steps, necessary for the successful execution of this research proposal, is presented in the progress report.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
7R01EY002708-14
Application #
3257086
Study Section
Clinical Sciences Study Section (CLN)
Project Start
1991-07-01
Project End
1993-02-28
Budget Start
1991-07-01
Budget End
1992-02-29
Support Year
14
Fiscal Year
1991
Total Cost
Indirect Cost
Name
University of Louisville
Department
Type
Schools of Medicine
DUNS #
City
Louisville
State
KY
Country
United States
Zip Code
40292
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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