1. F exam 2 F learning in a natural environment F special case... flower learning F odor learning in the proboscis extension reflex F summary PART 4: BEHAVIORAL PLASTICITY #19: ASSOCIATIVE LEARNING IN HONEYBEES II

2. F exam 2 F learning in a natural environment F special case... flower learning F odor learning in the proboscis extension reflex F summary PART 4: BEHAVIORAL PLASTICITY #19: ASSOCIATIVE LEARNING IN HONEYBEES II

3.  Dr. Spence Behmer F Department of Entomology, Texas A & M University F Friday, April 6 th @ 3:30 WHI Auditorium F Grasshoppers and nutritional physiology: from learning to community structure F to supplement your enjoyment or if you can’t make the seminar… literature will be posted F be ready for one bonus question on the final SEMINAR

4. F PER from sucrose contact on proboscis or antennae F reliable in hungry bees F studied in the laboratory F used with odor for conditioning F some simple rules for learning... ODOR LEARNING & PROBOSCIS EXTENTION REFLEX

5. F odor by itself does not elicit PER F classical or Pavlovian conditioning F US = sucrose F UR = PER F CS = odor F training: US + CS F test: CR = PER F bees learn that odor predicts sucrose reward ODOR LEARNING & PROBOSCIS EXTENTION REFLEX

6. F in color experiment F classical or Pavlovian conditioning F US = sucrose F UR = feeding F CS = color F training: US + CS F test: CR =  preference F bees learn that color predicts sucrose reward ODOR LEARNING & PROBOSCIS EXTENTION REFLEX

7. F CS ( e.g., color) must precede US (sucrose) ODOR LEARNING & PROBOSCIS EXTENTION REFLEX

8. F CS ( e.g., pattern) must precede US ( e.g., sucrose) F can use 2 CS stimuli = differential conditioning F CS + = paired with US F CS – = not paired with US F CR = shift in preference for either  training  testing ODOR LEARNING & PROBOSCIS EXTENTION REFLEX

9. F CS ( e.g., odor) must precede US ( e.g., shock) F can use 2 CS stimuli = differential conditioning F CS + = paired with US F CS – = not paired with US F CR = shift in preference for either odor “A” (CS + ) + shock (US) odor “B” (CS – ) flies  choice point  training  testing ODOR LEARNING & PROBOSCIS EXTENTION REFLEX

10. F CS ( e.g., odor) must precede US ( e.g., shock) F timing critical F determine interstimulus interval function ODOR LEARNING & PROBOSCIS EXTENTION REFLEX

11. F interstimulus interval function determination in PER F CS = geraniol odor F US = sucrose F optimal response when CS preceeds US by 2-3s ODOR LEARNING & PROBOSCIS EXTENTION REFLEX

12. F interstimulus interval function determination in PER F similar to color learning in free flying bees F CS = color F US = sucrose F optimal response when CS preceeds US by 2-3s ODOR LEARNING & PROBOSCIS EXTENTION REFLEX

13. F acquisition function determination in PER F how fast do bees learn? F CS = odor F US = sucrose F asymptotic response with 3 trials F excitatory or faciliatory conditioning leads to CR  ODOR LEARNING & PROBOSCIS EXTENTION REFLEX

14. F CS as predictor of the absence of US F how does unpaired CS pre-exposure influence CR? F CS = odor F US = sucrose F inhibitory conditioning reduces effect of training F CS is a less reliable predictor of US for the bees ODOR LEARNING & PROBOSCIS EXTENTION REFLEX

15. F differential conditioning in PER F can it be reversed? F CS + = odor A paired F CS – = odor B unpaired F US = sucrose F 1 st generalize to odor, then differentiate with training F CR + & CR – are opposite F reversal training = retrain bees to alternate odors ODOR LEARNING & PROBOSCIS EXTENTION REFLEX

16. F insect brains ~ vertebrate brains F fewer neurons F ~ complexity ANATOMY OF PER

17. F insect brains ~ vertebrate brains F fewer neurons F ~ complexity F e.g., Drosophila ANATOMY OF PER

18. F insect brains ~ vertebrate brains F fewer neurons F ~ complexity F e.g., Drosophila F sensory input ANATOMY OF PER

19. F insect brains ~ vertebrate brains F fewer neurons F ~ complexity F e.g., Drosophila F sensory input F motor output ANATOMY OF PER

20. F insect brains ~ vertebrate brains F fewer neurons F ~ complexity F e.g., Drosophila F sensory input F motor output F processing ANATOMY OF PER

21. F insect brains ~ vertebrate brains F fewer neurons F ~ complexity F e.g., Drosophila F sensory input F motor output F processing F neuropile F cell bodies ANATOMY OF PER

22. F honeybee brain F ~ 10 6 neurons F complex F what neuropiles mediate PER? ANATOMY OF PER

23. F honeybee brain F ~ 10 6 neurons F complex F what neuropiles mediate PER? F CS ANATOMY OF PER

24. F honeybee brain F ~ 10 6 neurons F complex F what neuropiles mediate PER? F CS F US + “VUM” ANATOMY OF PER

25. F honeybee brain F ~ 10 6 neurons F complex F what neuropiles mediate PER? F CS F US F CS + US convergence? ANATOMY OF PER

26. F CS + US convergence? F 3 regions tested using bilateral cooling: F antennal lobe F mushroom body  -lobe F mushroom body  -lobe ANATOMY OF PER

27. F CS + US convergence? F 3 regions tested using bilateral cooling: F antennal lobe F mushroom body  -lobe F mushroom body  -lobe ANATOMY OF PER

28. F CS + US convergence? F 3 regions tested using bilateral cooling: F antennal lobe F mushroom body  -lobe F mushroom body  -lobe F distributed function F MB most critical ANATOMY OF PER

29. F CS + US convergence? F two specific neurons tested for PER functions: F PE1 F VUMmx1 ANATOMY OF PER

30. F CS + US convergence? F PE1 F dye filled ANATOMY OF PER

31. F CS + US convergence? F PE1 F intracellular recordings F reduced response after training ANATOMY OF PER

32. F CS + US convergence? F PE1 F intracellular recordings F qualitative change in response with multiple trials F may be change in PE1 or change in KC input F suggest PE1 involvement in memory acquisition ANATOMY OF PER

33. F CS + US convergence? F VUMmx1 F ventral unpaired medial F dye filled F VERY extensive pattern of arborization F all areas of CS & US input ANATOMY OF PER

34. F CS + US convergence? F VUMmx1 F intracellular recording F fires in response to sucrose application F triggers neural circuit  proboscis extension* ANATOMY OF PER

35. F CS + US convergence? F VUMmx1 F substitute sugar with injected current F measurement of PER* F VUMmx1 provides US F current alone does  UR F separates sensory from US at neural level ANATOMY OF PER

36. F CS + US convergence? F VUMmx1 F intracellular recordings F  spikes with forward conditioning only ANATOMY OF PER

37. F CS + US convergence? F VUMmx1 F differential conditioning F intracellular recordings F responds to CS + only ANATOMY OF PER

38. F CS + US convergence? F VUMmx1 F 1 st order conditioning F train: CS1 (odor A) + US1 (sugar) F test: CS1  CR1 (PER) F 2 nd order conditioning F train: CS2 (odor B) + CS1 (odor A = US2) F test: CS2  CR2 (PER = CR1) ANATOMY OF PER

39. F CS + US convergence? F VUMmx1 F octopamine transmitter F substitute sugar with octopamine injection @ F calyx F antennal lobe F measurement of PER* F VUMmx1 provides US ANATOMY OF PER

40. F learning necessary for survival in nature F several forms of learning F several modalities of learning F rapid learning F permanent memory in 3 trials F temporal requirements of learning critical F waggle dance F integrate stimuli F can learn complex patterns F can learn general environmental features LEARNING IN HONEYBEES: SUMMARY

41. F proboscis extension reflex (PER) F show all features of learning described in vertebrates F temporal requirements of learning F differential conditioning F inhibitory learning F 2 nd order conditioning F structures correlated with learning F mushroom bodies & antennal lobes F neurons correlated with learning F PE1 & VUMmx1 LEARNING IN HONEYBEES: SUMMARY