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Lecture 20 : Olfaction II. 11/11/09. For next week. See web site Background - 1 pg. Topics. Diversity of ORs Which receptors detect which odorant? Olfactory signal transduction How does it compare to visual transduction? Multiple olfactory organs
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Lecture 20 : Olfaction II 11/11/09
For next week • See web site • Background - 1 pg
Topics • Diversity of ORs Which receptors detect which odorant? • Olfactory signal transduction How does it compare to visual transduction? • Multiple olfactory organs Nose vs VNO : How do the receptors compare? • Signal transduction How do pathways compare?
Functional response of rat I7 receptor I substitutions II chain length III/IV degree sat III/V side chain Response R>G>B
Functional group Compare response of control cells vs ones containing I7 receptor
Length and saturation C8 is best though receptor responds to others too
Expressing olfactory receptors • Ways to express genes • Make transgenic animals • Express in Xenopus oocytes • Ideal to do in cell culture • Ways to probe • Intracellular recording of one cell • Pull off patch of membrane • Ca+2 imaging
Olfactory signal transduction Odorant in activates receptor activates Golf activates AC makes cAMP
Monitor cell response with luciferase CRE luciferase CRE = cAMP response element luciferin Odorant in -> cAMP -> luciferase + luciferin = LIGHT
Screen panel of 96 receptors Flow reactant 1 and determine which receptors respond
Screen panel of 96 receptors Flow reactant 2 and determine which receptors respond Map out response space
OR response • Map out response to range of chemicals and odorant levels • Different receptors respond to different ligands
Human perceptual variation • Androstenone is odorous steroid • Derived from testosterone • Human response varies • Offensive = Sweat / urine • Pleasant = sweet, floral • Odorless
OR7D4 receptor is sensitive for androstenone/androstadienone Screened 335 OR’s (85% of known ORs!)
Human allelic variation in OR7D4 • Search SNP database and seq in 391 test subjects - variation at 2 AA • R88W T133M • Two common genotypes RT or WM • Normal (RT) 0.786 • WM 0.157 • P79L 0.04 • S84N 0.013
WM allele much less sensitive than RT allele RT vs WM allele
Rare alleles have opposite effects P79L not sensitive S84N very sensitive Chimp even more sensitive
Olfactory signal recovery 1. OR* gets phophorylated and binds arrestin 2. PDE turns cAMP into AMP This decreases cAMP binding to channel Channel closes #1 #2
Olfactory signal recovery Adaptation CaM-Ca+2 binds to channel and decreases cAMP binding CaM-kinase phosphorylates PDE which acts faster CaM kinase phosphorylates AC which acts slower
Multiple olfactory cues • Need to detect general odors • Food, environment • Species specific odors • Pheromone = • pherein (to carry or transfer) + • horman (to excite) • Volatile (broadcast info) or non-volatile (individual specific)
Organs for detecting compounds • Olfactory • Vomeronasal • Trigeminal
Mouse olfaction Main olfactory epithelium (MOE) Expresses olfactory receptors OR (1300) and Trace amine associated receptors TAAR Project to main olfactory bulb (MOB) Brennan and Zufall 2006
Mouse olfaction Vomeronasal organ (VNO) Expresses two types of receptors: V1R (137) and V2R (60) Project to Accessory olfactory bulb (AOB) Brennan and Zufall 2006
VNO Two kinds of receptors which differ in location and projection Response to putative pheremones
Diversity of GPCRs Bargmann 2006
G • Gs stimulates adenylate cyclase • includes olfactory • Gi/Go • Includes vision and taste • Gq • Activates PLC V2R Vision/taste V1R OR
VNO mystery #1 • Are vomeronasal receptors for species specific communication?
VNO receptors Species specific as if they evolved for intra-species communication