1. For every change in a given behavior there must be an underlying change in the neural circuitry that mediates that change Mechanisms of control of plasticity: Development/genes imprinting language acquisition song learning Environment associations (classical and instrumental learning) lack of associations (habituation) Neural and behavioral plasticity

2. Olfactory learning in Manduca sexta

3. Antennal lobe anatomy: analogue of the vertebrate olfactory bulb

4. Development of an olfactory-learning behavioral paradigm EMG electrode in feeding muscle exposed proboscis tip for sucrose application reference electrode odor blown across antennae Daly and Smith J Exp. Biol (2000) Moths normally do not respond to odor with a feeding response Moths innately respond to sugar water with a feeding response, which can be measured with electromyography Prediction: If odor predicts food AND moths learn then moths should produce an anticipatory feeding response to odor if odor and food are paired. Measures: Change in feeding muscle activity Extension of the proboscis

5. Assessing the contribution of associative and non associative effects A. C. B. odor (CS) food (US) on off CS US CS US forward-paired and air-paired CS-US relationship backward-paired CS-US relationship random-paired CS-US relationship on -25sec +25sec 4 sec

6. A. prepost 05 10 B. prepost 0510 D. prepost 05 10 Time (sec) C. prepost 05 10 Time (sec) Characteristic EMG Responses Pre-conditioningAfter forward-pairing 24+ h after Forward-pairing After backward and random- pairing odor on

7. Acquisition of conditioned response 312654.80.70.60.50.40.30.20 0.10 Geraniol Conditioning Trials Response probability forward pairing random pairing air forward pairing N=40/group

8. air- forward pairing TRIAL forward pairing backward pairing random pairing BEFORE TRIAL 1 BETWEEN TRIALS 5 & 6 AFTER TRIAL 10 CHANGE IN EMG SPIKE FREQUENCY (POST-PRE) 0 1 2 3 4 5 7 8 6 10 9 Test Trial (EMG Response) Only forward pairing produces a conditioned response *N=30 per group

9. How does this learning influence behavior in free flying moths?

10. In-flight odor response of conditioned moths to conditioning odor Mean post-conditioning wind tunnel odor response:Forward-paired vs Odor-only 0 20 40 60 Odor- mediated Flight Reach Source Proboscis Extension at Odor Source Observed behavior % Response Forward-paired Odor-only N=20/group

11. 80 70 60 50 40 30 20 10 0 Percent CR cyclohexanone methyl jasmonate 12345 6 Olfactory learning is conditional: biologically meaningful odors resist learning N=40/group MJ –/CH +CH –/MJ + Delta spike frequency 14 12 10 8 6 4 2 0 -2 N=40/group 14 12 10 8 6 4 2 0 -2 MS –/CH +CH –/MS + Delta spike frequency N=40/group

12. Olfactory learning can be used to assay ability: Generalization of CR to odors of increasing difference Conditioning odor Odor dimension (carbon chain length) Generalization gradient | Similar odors || Different odorsDifferent odors | Response probability

13. .60.50.40.30.20.10 0 123456 Conditioning Trials Response probability 2-hexanone 1-hexanol 1-decanol Acquisition of CR to the conditioning odor N=80/group

14. 1-Hexanol 1-Heptanol 2-Heptanone 1-Octanol 2-Octanone 1-Decanol 2-Hexanone Cyclohexanone.80.70.60.50.40.30.20.10 0 Response probability 1-Hexanol 1-Heptanol 2-Heptanone 1-Octanol 2-Octanone 1-Decanol 2-Decanone Cyclohexanone Response probability.80.70.60.50.40.30.20.10 0 2-Hexanone 1-Heptanol 2-Heptanone 1-Octanol 2-Octanone 1-Decanol 2-Decanone Cyclohexanone Response probability.80.70.60.50.40.30.20.10 0 CS ketone alcohol cyclohexanone Generalization of the CR to conditioning and test odors N=80/group

15. Inhibitory gradient Summation of + and – gradients along a single odor dimension Summed inhibitory and excitatory gradients Excitatory generalization gradient Theoretical odor dimension

16. .60.50.40.30.20.10 0 12 34 56 Conditioning Trials Response probability 1-octanol (A) 1-hexanol (B).80.70.60.50.40.30.20.10 0 1-Heptanol 1-Octanol 1-Decanol 1-Hexanol 1-nonanol Response probability (CS+) (CS-) Differential conditioning produces + and – gradients which overlap along a single dimension and summate in predictable ways.

17. .60 50.40.30.20.10 0 12 34 56 Conditioning Trials A. Response probability 1-heptanol (+) 1-hexanol (-) 1-Heptanol 1-Octanol 1-Decanol 1-Hexanol.80.70.60.50.40.30.20.10 0 (+) (-) B. Response probability a b a a Differential conditioning produces + and – gradients which overlap along a single dimension and summate in predictable ways.

18. Learning related change in the Al implies feedback