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Discoveries of odorant receptors and the organization of the olfactory system

The Nobel Prize in Physiology or Medicine 2004. Discoveries of odorant receptors and the organization of the olfactory system. 醫學一 B9602084 潘 岳 B9602091 謝仁峰 B9602086 鄭尉佐 B9602098 羅緯麟 B9602088 盧柏嘉.

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Discoveries of odorant receptors and the organization of the olfactory system

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  1. The Nobel Prize in Physiology or Medicine 2004 Discoveries of odorant receptors and the organization of the olfactory system 醫學一 B9602084 潘 岳 B9602091 謝仁峰 B9602086 鄭尉佐B9602098 羅緯麟 B9602088 盧柏嘉

  2. Discoveries of odorant receptors and the organization of the olfactory system Richard Axel Linda B.Buck Fred Hutchinson Cancer Research Center Seattle, WA, USA Columbia University New York, NY, USA

  3. Three assumptions Receptors are G-protein-coupled receptors. (GPCRs) Diverse receptors encoded by multigene family Receptor expression restricted to olfactory epithelium

  4. Homogenize the epithelium cells ‧RNA

  5. Many double-stranded cDNAs • Reverse transcriptase • cDNA • Specific primer • GPCRs family • PCR Add RNA andreverse transcriptase primer

  6. Add primers anneal to conserved regions(II VII) of the GPCR family VII II

  7. Different species cDNA exist

  8. Electrophoresis：initiation M 3600 1700 1500 1200 900 600 300

  9. Electrophoresis：result M 3600 1700 1500 1200 900 600 300

  10. Add restriction enzyme Simple cDNAs Mixed-up cDNAs M 3600 1700 1500 1200 900 600 300

  11. Northern blot Material RNA from epithlium/brain/spleen labeled PCR13 products as probe Result Brain & Spleen :no hybridization Expression is restricted to olfactory epithelium Demonstrate assumption 3 Obtain full-length c-DNA

  12. Protein sequence

  13. N-terminus various Related to ligand recognition C-terminus

  14. Experiment summary • DNA sequencing Assumption 1 :GPCR Assumption 2 :Multigene→various receptor • Northern blotting Assumption 3 :Expression location protein sequence

  15. The family of olfactory genes in human Over 1000 genes About 400 genes function ; 600 genes not function

  16. Olfactory receptor genes

  17. Olfactory sensory neuron

  18. High Low Sensitivity Species differences

  19. Olfactory Sensory Neuron(OSN) Olfactory Sensory neuron

  20. A large family of odorant receptors • One neuron cell expresses only one type of receptor • 7 transmenbrane domains • Nitrite group in extracellular space • Conformational change Olfactory Sensory neuron

  21. Combinatorial receptor codes

  22. Sensory transduction

  23. Sensorytransduction

  24. Olfactory Bulb

  25. Olfactory Receptor V.S. Olfactory glomerulus

  26. Map of Olfactory Bulb

  27. 前 後

  28. Q：Human has about 400 types of receptors, but how do they discriminate about ten thousands odorants?

  29. Combinatorial receptor codes experiment 100 10 1 Fura-2

  30. Combinatorial receptor codes

  31. Different ordor recognized Multigenefamily Functional genes：pseudogenes 400：600 express Receptor Combination

  32. Thank You！

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