The long-term goal of this research is to understand from a cell biological perspective how neurons establish and maintain distinct axonal and dendritic domains that differ in structure and function. This feature of nerve cells-referred to as neuronal polarity- is essential to normal neural function. Disruption of neuronal polarity is thought to contribute to the pathophysiology of many neurologic diseases.
The aims for the coming award period are to elucidate the cellular mechanisms underlying the targeting and transport of membrane proteins and to assess the role that these processes play in the development of neuronal polarity. In the previous award period it was shown that basolateral targeting signals that contain a tyrosine motif target exogenous proteins to dendrites, where exogenous proteins carrying motifs in the cytoplasmic domain of endogenous dendritic protein mediate their targeting and will identify the sequence within an endogenous axonal protein that govern its polarization to the axon. The earliest event in the establishment of neuronal polarity occurs when one of several, initially identical processes undergoes a prolonged period of growth, becoming the cell's axon. In order to assess possible causal relationships between specification of the axon and the development of protein targeting, changes in the distribution of representative axonal and dendritic proteins labeled with Green Fluorescent Protein will be followed as cells begin axonal outgrowth. Further experiments will perturb the localization of an endogenous cell adhesion molecule involved in the signaling that governs the specification of polarity in order to assess its impact on axonal outgrowth and the development of polarity. After the initial sorting event, which is presumed to direct different proteins to distinct vesicle populations, transport vesicles must be targeted to appropriate domains within the cell. The expression of appropriately tagged axonal, dendritic, and uniformly distributed proteins will be used to define the populations of transport vesicles present in hippocampal neurons and to visualize their transport in living cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS017112-19
Application #
6126090
Study Section
Neurology B Subcommittee 2 (NEUB)
Program Officer
Chiu, Arlene Y
Project Start
1981-08-01
Project End
2002-11-30
Budget Start
1999-12-01
Budget End
2000-11-30
Support Year
19
Fiscal Year
2000
Total Cost
$304,393
Indirect Cost
Name
Oregon Health and Science University
Department
Neurosciences
Type
Schools of Medicine
DUNS #
009584210
City
Portland
State
OR
Country
United States
Zip Code
97239
Kaech, Stefanie; Huang, Chun-Fang; Banker, Gary (2012) Short-term high-resolution imaging of developing hippocampal neurons in culture. Cold Spring Harb Protoc 2012:340-3
Kaech, Stefanie; Huang, Chun-Fang; Banker, Gary (2012) Long-term time-lapse imaging of developing hippocampal neurons in culture. Cold Spring Harb Protoc 2012:335-9
Kaech, Stefanie; Huang, Chun-Fang; Banker, Gary (2012) General considerations for live imaging of developing hippocampal neurons in culture. Cold Spring Harb Protoc 2012:312-8
Davare, Monika A; Fortin, Dale A; Saneyoshi, Takeo et al. (2009) Transient receptor potential canonical 5 channels activate Ca2+/calmodulin kinase Igamma to promote axon formation in hippocampal neurons. J Neurosci 29:9794-808
Soderling, Scott H; Guire, Eric S; Kaech, Stefanie et al. (2007) A WAVE-1 and WRP signaling complex regulates spine density, synaptic plasticity, and memory. J Neurosci 27:355-65
Oliva Jr, Anthony A; Atkins, Coleen M; Copenagle, Lily et al. (2006) Activated c-Jun N-terminal kinase is required for axon formation. J Neurosci 26:9462-70
Kaech, Stefanie; Banker, Gary (2006) Culturing hippocampal neurons. Nat Protoc 1:2406-15
Withers, Ginger S; James, Conrad D; Kingman, Caroline E et al. (2006) Effects of substrate geometry on growth cone behavior and axon branching. J Neurobiol 66:1183-94
Jacobson, Catherine; Schnapp, Bruce; Banker, Gary A (2006) A change in the selective translocation of the Kinesin-1 motor domain marks the initial specification of the axon. Neuron 49:797-804
Silverman, M A; Peck, R; Glover, G et al. (2005) Motifs that mediate dendritic targeting in hippocampal neurons: a comparison with basolateral targeting signals. Mol Cell Neurosci 29:173-80

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