Both neurons and glia of the PNS are derived from the neural crest. In this study, we have examined the potential function of lunatic fringe in neural tube and trunk neural crest development by gain-offunction analysis during early stages of nervous system formation. Normally lunatic fringe is expressed in three broad bands within the neural tube, and is most prominent in the dorsal neural tube containing neural crest precursors. Using retrovirally-mediated gene transfer, we find that excess lunatic fringe in the neural tube increases the numbers of neural crest cells in the migratory stream via an apparent increase in cell proliferation. In addition, lunatic fringe augments the numbers of neurons and upregulates Delta-1 expression. The results indicate that, by modulating Notch/Delta signaling, lunatic fringe not only increases cell division of neural crest precursors, but also increases the numbers of neurons in the trunk neural crest.
This laboratory has been using a derivatized bead assay to study cell surface properties in many experimental systems. Here we test its efficacy in the yeast/concanavalin A (Con A) binding model to determine if immobilized Con A is inhibited by the same saccharides that inhibit free Con A binding. This system offers an excellent test case for the efficacy and validity of the derivatized bead assay. Con A binds to yeast because yeast are rich in cell surface mannose residues, a Con A preferential binding sugar. Here we examine the effects of 30 different sugars (0.05 M) on the binding of yeast to agarose beads derivatized with Con A, in a total of 3998 trials, with an average of over 133 replicates for each sugar. The most inhibitory sugars included D(+) melezitose, D(+) trehalose and maltotriose, the same sugars that effectively inhibit binding of free Con A. The results suggest that the bead assay is an effective approach to study the binding properties of cells, and most important, it is so rapid that hundreds of trials can be done in the time it would take to do one trial using conventional assays (supported by NIH NIGMS SCORE, RISE, MARC, the ITQ program, and the Joseph Drown Foundation).
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