The overall goal of this research is to use an in vitro model system, developed in our laboratory, to study the molecular mechanism of glial-guided neuronal migration in the developing brain. The proposed research will purify and characterize a novel glycoprotein, ASTROTACTIN, antibodies against which block neuron-glial interactions in the microculture system. Monoclonal and mono-specific polyclonal anti-astrotactin antibodies, selected with in vitro functional assays, will be used to purify astrotactin, to select cDNA clones from a mouse lambda gtll expression library and to isolate fusion proteins from immunopositive clones. High resolution video microscopy will then be combined with computer-assisted image processing to visualize the effects of Fab fragments of mono-specific astrotactin antibodies on the interaction of the migrating neuron and its leading process with the glial fiber. Biochemical studies on astrotactin and its proteolytic cleavage products will be used to analyze the size, shape and functional domains of the molecule. This information will be combined with in vitro assays of the effects of astrotactin peptides, fusion peptides and monoclonal antibodies to define the regions of astrotactin that regulate neuron-glial interactions. We will then use immunocytochemical and molecular biological methods to study the expression of astrotactin in the developing C57BL/6J mouse brain and in the weaver mouse mutant mouse, because we have found that astrotactin is missing or defective in the medial aspect of the weaver cerebellum. Finally, we will raise anti-idiotypic antibodies against anti-astrotactin antibodies and use affinity chromatography to attempt to purify a receptor for astrotactin from glial cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS015429-12
Application #
3396251
Study Section
Neurology B Subcommittee 1 (NEUB)
Project Start
1987-09-01
Project End
1991-02-28
Budget Start
1989-09-01
Budget End
1991-02-28
Support Year
12
Fiscal Year
1989
Total Cost
Indirect Cost
Name
Columbia University (N.Y.)
Department
Type
Schools of Medicine
DUNS #
064931884
City
New York
State
NY
Country
United States
Zip Code
10027
Schneider, Stephanie; Gulacsi, Alexandra; Hatten, Mary E (2011) Lrp12/Mig13a reveals changing patterns of preplate neuronal polarity during corticogenesis that are absent in reeler mutant mice. Cereb Cortex 21:134-44
Wilson, Perrin M; Fryer, Robert H; Fang, Yin et al. (2010) Astn2, a novel member of the astrotactin gene family, regulates the trafficking of ASTN1 during glial-guided neuronal migration. J Neurosci 30:8529-40
Osheroff, Hilleary; Hatten, Mary E (2009) Gene expression profiling of preplate neurons destined for the subplate: genes involved in transcription, axon extension, neurotransmitter regulation, steroid hormone signaling, and neuronal survival. Cereb Cortex 19 Suppl 1:i126-34
Solecki, David J; Trivedi, Niraj; Govek, Eve-Ellen et al. (2009) Myosin II motors and F-actin dynamics drive the coordinated movement of the centrosome and soma during CNS glial-guided neuronal migration. Neuron 63:63-80
Zhao, Haotian; Ayrault, Olivier; Zindy, Frederique et al. (2008) Post-transcriptional down-regulation of Atoh1/Math1 by bone morphogenic proteins suppresses medulloblastoma development. Genes Dev 22:722-7
Solecki, David J; Govek, Eve-Ellen; Tomoda, Toshifumi et al. (2006) Neuronal polarity in CNS development. Genes Dev 20:2639-47
Solecki, David J; Govek, Eve-Ellen; Hatten, Mary E (2006) mPar6 alpha controls neuronal migration. J Neurosci 26:10624-5
Hatten, Mary E (2005) LIS-less neurons don't even make it to the starting gate. J Cell Biol 170:867-71
Hatten, Mary E; Heintz, Nathaniel (2005) Large-scale genomic approaches to brain development and circuitry. Annu Rev Neurosci 28:89-108
Millen, K J; Millonig, J H; Hatten, M E (2004) Roof plate and dorsal spinal cord dl1 interneuron development in the dreher mutant mouse. Dev Biol 270:382-92

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