Dale D. Hunter - US grants

Environmental cues play a major role in determining the ultimate fate of the developing nervous system. Our goals are to identify some of the signals recognized by developing neurons as they migrate, differentiate, and form synaptic connections. Previously, as a postdoctoral fellow in the laboratory of Joshua Sanes at Washington University, I identified a component of the synaptic basal lamina of the neuromuscular junction, s-laminin, that is likely to be involved in the stabilization of the mature motor synapse. I obtained antibodies to and cDNA clones for this extracellular matrix molecule, and we are currently developing similar resources for other potentially neuroactive molecules. We plan on using both previously and newly generated antibodies as immunohistochemical and immunopurification tools for identifying the location and function of matrix molecules in the developing nervous system. Similarly, we plan on using previously and newly generated cDNA probes to examine the expression and regulation of these molecules during development. We propose to use immunohistochemical and molecular biological methods to determine where and when s-laminin is expressed in the developing cerebral cortex and olfactory system, and to determine the function of s-laminin in these areas during development, using both in vitro and in vivo systems. In addition, we propose to identify the neural receptor for s-laminin, using purified s-laminin and s-laminin-derived peptides as affinity reagents. Finally, we propose to identify and characterize other matrix components that may be important determinants of development in the central nervous system, using both immunohistochemical and molecular biological techniques. Together, these lines of investigation should help to define the role of extracellular matrix molecules in the development and differentiation of the nervous system.

Environmental cues play a major role in determining the ultimate fate of the developing nervous system. Our goals are to identify some of the signals recognized by developing neurons as they migrate, differentiate, and form synaptic connections. Previously, as a postdoctoral fellow in the laboratory of Joshua Sanes at Washington University, I identified a component of the synaptic basal lamina of the neuromuscular junction, s-laminin, that is likely to be involved in the stabilization of the mature motor synapse. I obtained antibodies to and cDNA clones for this extracellular matrix molecule, and we are currently developing similar resources for other potentially neuroactive molecules. We plan on using both previously and newly generated antibodies as immunohistochemical and immunopurification tools for identifying the location and function of matrix molecules in the developing nervous system. Similarly, we plan on using previously and newly generated cDNA probes to examine the expression and regulation of these molecules during development. We propose to use immunohistochemical and molecular biological methods to determine where and when s-laminin is expressed in the developing cerebral cortex and olfactory system, and to determine the function of s-laminin in these areas during development, using both in vitro and in vivo systems. In addition, we propose to identify the neural receptor for s-laminin, using purified s-laminin and s-laminin-derived peptides as affinity reagents. Finally, we propose to identify and characterize other matrix components that may be important determinants of development in the central nervous system, using both immunohistochemical and molecular biological techniques. Together, these lines of investigation should help to define the role of extracellular matrix molecules in the development and differentiation of the nervous system.

DESCRIPTION (Adapted from applicant's abstract): The development of the nervous system is modulated by both genetic and environmental phenomena. Whereas intrinsic factors undoubtedly play roles in the potential of progenitor cells to differentiate into a given cell type, extrinsic clues are likely to serve as signals for the timing of these differentiative events. The proposed project is designed to examine the effects of environmental factors on a cell's phenotype, particularly the environmental factors present in the extracellular matrix (ECM). Experiments are proposed to identify the distribution, expression, and function of components of the extracellular matrix, in particular, the beta2-containing laminins, during the development of the retina using antibodies and cDNA probes, both in vitro and in vivo. Together, it is anticipated that these lines of investigation should help to define the role of these extracellular matrix molecules in the development of the nervous system.

This Core will provide Neuroscience investigators the statistical and bioinformatics tools required for the analysis of gene expression data. This Core takes advantage of the existing recently established T-NEMC Expression Array Core, that creates and analyses DNA arrays. Dr. Dale Hunter, who currently oversees the T-NEMC Expression Array Core, will direct this Core. The personnel for the Core consist of Dr. Wollenberg a bioinformatics expert (10% effort), and a to be hired bioinformatics technician (100%). In addition, the Core will purchase licenses for a sophisticated commercial software package, GeneSpring, ideal for analysis of scanned expression array image data and subsequent data mining. The addition of the Bioinformatics Core to the services provided by TEAC will allow NINDS funded investigators to receive a reduced fee structure for gene array core services.