Odor Receptors Govern the Formation of a Precise 气味受体支配一个精确的形成.pptVIP

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Odor Receptors Govern the Formation of a Precise 气味受体支配一个精确的形成.ppt

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Odor Receptors Govern the Formation of a Precise 气味受体支配一个精确的形成

Odor Receptors Govern the Formation of a Precise Topographic Map Department of Biochemistry and Molecular Biophysics Columbia University New York, NY 10032 Wang, F., Nemes, A., Mendelsohn, M., Axel, R. Odorant receptor proteins in olfactory receptor neurons are key components in guiding axon terminal location. This means that the receptors help to provide a topographic map within the olfactory bulb that distinguishes odorants. Construction of P2-deletion plasmids Generation of targeted mutations P2-Deletion-IRES-tau-lacZ M71?P2-IRES-tau-lacZ M50?P2-IRES-tau-lacZ P3?P2-IRES-tau-lacZ Immunohistochemistry to detect β-galactosidase was performed. 28 μm coronal sections were dissected and frozen in OCT Tissues were fixed and then reacted with anti-β-galactosidase antibody at 1:1000 dilution in PTS The bound antibody was visualized using Cy3-conjugated anti-rabbit IgG X-gal staining was performed Methods Results – P2-Deletion P2 ORNs converge on exactly 2 out of 1800 glomeruli (lateral and medial) P2-Deletion-IRES-tau-lacZ mutants pass through cribiform plate into OB but do not converge onto glomeruli Results – M71 substitution M71 receptor gene resides in different chromosome and different epithelial zone than P2 (radical substitution) Axons of wild-type M71 receptor converge on opposite end of OB compared to P2 axons ORNs expressing M71?P2-IRES-tau-lacZ allele still reside in P2 zone in epithelium ORNs expressing M71?P2-IRES-tau-lacZ allele converge on glomeruli other than M71 or P2 Results – M50 Substitution M50 gene located on same chromosome (#7) as P2 gene – however different epithelial zone ORNs expressing M50?P2-IRES-tau-lacZ allele reside in P2 zone in epithelium. ORNs expressing M50?P2-IRES-tau-lacZ allele project axons to glomeruli distinct from either M50 or P2 wildtype Results – P3 substitution P3 gene located on same chromosome and same epithelial zone as P2 P3 glomerulus lies 200 um anterior to P2 glomerulus (about 3 glomeruli separation) ORNs expressing