Neurofilaments are major proteins of the neuronal cytoskeleton, although their function is not well understood at present. They are unusual, compared to other intermediate filaments, in that they express considerable quantities of very heavily phosphorylated high molecular weight material, some of which is apparently associated with cross-bridging structures. I have demonstrated clear immunological differences between neurofilaments in axons and dendrites and between those of mature and developing regions of the nervous system. Such distributions suggest participation of neurofilaments in the mechanisms of development, plasticity and axon/dendrite differentiation. In addition changes in neurofilamentous organization are characteristic of common human pathologies, and may be implicated in Alzheimers disease.
The aims of this grant are 1. to examine the extent and significance of neurofilament phosphorylation using immunocytochemical approaches, and to assess the importance of this modification in the production of the immunological differences observed; 2. to identify, purify and characterize protein kinases responsible for this phosphorylation; 3. to study the cell type distribution of these kinases within the developing and mature nervous system; 4. to examine the relevance of neurofilament phosphorylation in the formation and control of neurofilament cross-links; 5. to assess the importance of neurofilament phosphorylation directly by microinjection of function blocking antibodies into cultured neurones; 6. to assess the effect of drugs known to cause neurofilamentous accumulations in vivo on the growth and morphology of cultured neurones; 7. to examine the effect of these drugs on described in vitro neurofilament-microtubule interactions. The work proposed will lead to an understanding of the control, extent and function of neurofilament phosphorylation and will test my hypothesis that drugs inducing neurofilamentous neuropathies act primarily on the heavily phosphorylated high molecular weight neurofilament proteins.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
1R01NS022695-01
Application #
3405463
Study Section
Neurological Sciences Subcommittee 1 (NLS)
Project Start
1985-07-01
Project End
1989-01-31
Budget Start
1985-07-01
Budget End
1987-01-31
Support Year
1
Fiscal Year
1985
Total Cost
Indirect Cost
Name
University of Florida
Department
Type
Schools of Medicine
DUNS #
073130411
City
Gainesville
State
FL
Country
United States
Zip Code
32611
Errante, L; Tang, D; Gardon, M et al. (1998) The intermediate filament protein peripherin is a marker for cerebellar climbing fibres. J Neurocytol 27:69-84
Lu, D; Yang, H; Shaw, G et al. (1998) Angiotensin II-induced nuclear targeting of the angiotensin type 1 (AT1) receptor in brain neurons. Endocrinology 139:365-75
Wang, D S; Deng, T; Shaw, G (1997) Membrane binding and enzymatic activation of a Dbl homology domain require the neighboring pleckstrin homology domain. Biochem Biophys Res Commun 234:183-9
Shaw, G; Miller, R; Wang, D S et al. (1997) Characterization of additional casein kinase I sites in the C-terminal ""tail"" region of chicken and rat neurofilament-M. J Neurochem 69:1729-37
Wang, D S; Miller, R; Shaw, R et al. (1996) The pleckstrin homology domain of human beta I sigma II spectrin is targeted to the plasma membrane in vivo. Biochem Biophys Res Commun 225:420-6
Benson, D L; Mandell, J W; Shaw, G et al. (1996) Compartmentation of alpha-internexin and neurofilament triplet proteins in cultured hippocampal neurons. J Neurocytol 25:181-96
Shaw, G (1996) The pleckstrin homology domain: an intriguing multifunctional protein module. Bioessays 18:35-46
Hollander, B A; Bennett, G S; Shaw, G (1996) Localization of sites in the tail domain of the middle molecular mass neurofilament subunit phosphorylated by a neurofilament-associated kinase and by casein kinase I. J Neurochem 66:412-20
Wang, D S; Shaw, G (1995) The association of the C-terminal region of beta I sigma II spectrin to brain membranes is mediated by a PH domain, does not require membrane proteins, and coincides with a inositol-1,4,5 triphosphate binding site. Biochem Biophys Res Commun 217:608-15
Wang, D S; Shaw, R; Hattori, M et al. (1995) Binding of pleckstrin homology domains to WD40/beta-transducin repeat containing segments of the protein product of the Lis-1 gene. Biochem Biophys Res Commun 209:622-9

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