https://doi.org/10.5061/dryad.tb2rbp098

The PI (preference index) for the vanilla odor of individual crickets is described for each experiment.

Description of the data and file structure

We developed datasets to evaluate the effect of various types of conditioning in crickets for addressing a question of whether integration of competing appetitive and aversive information takes place during the encoding of the experience or during memory retrieval [1, 2]. This study is based on our accumulated knowledge on the roles of octopamine neurons and dopamine neurons in formational and retrieval of memory in Pavlovian conditioning and second-order conditioning in crickets. [3-10]. Relative preference between the conditioned odor or visual pattern and the control odor or pattern was collected using test apparatuses [3, 4] at each step of conditioning trainings in crickets. Relative preference was defined as the preference index (PI) for the conditioned odor or visual pattern, defined as Tc/ (Tc+Tn), where Tc was the time spent exploring the odor or pattern used in conditioning and Tn was the time spent exploring the odor or pattern not used in conditioning in the test. The name of each dataset (Data Figs. 2a, 2b, 2c, 2d, 3a, 3b, 4a, 4b, 4c, S2a, S2b) is the name of the figure of our associated paper. The experiments for each data are as follows:

Data Fig. 2a-d. We investigated retention of appetitive and aversive memories after conditioning of an odor with water and then with sodium chloride solution as evaluated by injection of epinastine (OA receptor antagonist) or flupentixol (DA receptor antagonist). The time schedules of the experiments are shown above the graphs. (a, b) Two groups of crickets received appetitive and aversive conditioning for the same odor (CS). Odor preference was tested before (Pre-test), 30 min after appetitive conditioning (Post-test 1) and 30 min and 24 hours after aversive conditioning (Post-tests 2 and 3). Each test took 85-90 min. Crickets in each group were then injected with epinastine (a) or flupentixol (b) and received the final test 30 min later (Post-test 4). (c, d) Two groups of crickets received appetitive and aversive conditioning. Odor preference was tested before (Pre-test), 30 min after appetitive conditioning (Post-test 1) and 30 min after aversive conditioning (Post-test 2). Then the crickets in each group were injected with epinastine (c) or flupentixol (d) and received the final test 30 min later (Post-test 3). The result of this experiment demonstrates formation and expression of parallel memories after sequential appetitive and aversive conditioning of a CS.  In addition, the experiment demonstrates that expression of previously formed appetitive memory is transiently inhibited after subsequent aversive conditioning of the same CS.

Data Fig. 3a, b. We investigated formation of appetitive and aversive memories by pairing a visual pattern (CS2) with an odor (CS1) that was paired with water and then with sodium chloride solution, evaluated by injection of epinastine (a) or flupentixol (b). The time schedules of the experiments are shown above the graphs. Four groups of crickets received pairing of an odor (CS1) with water and then sodium chloride solution, and they received pairing of a pattern (CS2) with the odor (CS1) 24 hours later. Then the crickets in each group were injected with either saline (a, b) or saline containing (a) epinastine or flupentixol (b). Pattern preference was tested before training (Pre-test), 1 hour after CS2-CS1 pairing (Post-test 1) and 30 min after injection (Post-test 2). The results of this study indicate that appetitive and aversive memories were formed simultaneously for CS2 by pairing CS2 with CS1. In other words, appetitive and aversive SOCs are achieved at the same time by this pairing.

Data Fig. 4a, b. We investigated reduction of the expression of appetitive memory by US (water) devaluation after pairing of an odor (CS1) with water and then with sodium chloride solution (a, b) and after pairing of a visual pattern (CS2) with the odor (CS1) (c), evaluated by injection of epinastine or flupentixol. The time schedules of the experiments are shown above the graphs. (a, b) Crickets in four groups received an odor preference test 2 hours before training (Pre-test), 30 min after providing water until satiation (Post-test 1), and 30 min after injection of saline (a, b) or saline containing epinastine (a) or flupentixol (b) (Post-test 2). (c). Crickets in another two groups received a pattern preference test before training (Pre-test), 1 hour after pairing of a pattern (CS2) with the odor (CS1) paired with water and sodium chloride solution (Post-test 1), 1 hour after providing water until satiation (Post-test 2), and 30 min after injection of saline or flupentixol (Post-test 3). The results of this experiment indicate that expression of appetitive memory is inhibited in water-satiated crickets and hence aversive memory governs the response to the CS2.

Data Fig. S2a, b. We investigated retention of aversive and appetitive memories after aversive-to-appetitive counterconditioning (a, b) Crickets in two groups received pairing of an odor with sodium chloride solution and then with water. Odor preference was tested before (Pre-test), 30 min after aversive conditioning (Post-test 1) and 30 min and 24 hours after appetitive conditioning (Post-tests 2 and 3). The crickets were then injected with epinastine (a) or flupentixol (b) and received the final test 30 min later (Post-test 4). The results of this experiment demonstrate that aversive-to-appetitive counterconditioning is achieved.

By evaluating the dataset, we conclude that that appetitive and aversive conditioning systems operate independently to form parallel appetitive and aversive memories, which compete to produce learned behavior in crickets.

References

[1] Das G, Klappenbach M, Vrontou E, Perisse E, Clark CM, Burke CJ, Waddell S. 2014 Drosophila learn opposing components of a compound food stimulus. Curr. Biol. *24*, 1723-1730.

[2] Toshima N, Weigelt MK, Weiglein A, Boetzl FA, Gerber B. 2019 An amino- acid mixture can be both rewarding and punishing to larval Drosophila melanogaster. J. Exp. Biol. 222, jeb209486.

[3] Unoki S, Matsumoto Y, Mizunami M. 2005 Participation of octopaminergic reward system and dopaminergic punishment system in insect olfactory learning revealed by pharmacological study. Eur. J. Neurosci. 22, 1409-1416.

[4] Unoki S, Matsumoto Y, Mizunami M. 2006 Roles of octopaminergic and dopaminergic neurons in mediating reward and punishment signals in insect visual learning. Eur. J. Neurosci. 24, 2031-2038.

[5] Nakatani Y, Matsumoto Y, Mori Y, Hirashima D, Nishino H, Arikawa K, Mizunami M. 2009 Why the carrot is more effective than the stick: Different dynamics of punishment memory and reward memory and its possible biological basis. Neurobiol. Learn. Mem. 92, 370-380.

[6] Terao K, Matsumoto Y, Mizunami M. 2015 Critical evidence for the prediction error theory in associative learning. Sci. Rep. 5, 8929.

[7] Terao K, Mizunami M. 2017 Roles of dopamine neurons in mediating the prediction error in aversive learning in insects. Sci. Rep. 7, 14694.

[8] Awata H, Wakuda R, Ishimaru Y, Matsuoka Y, Terao K, Katata S, Matsumoto Y, Hamanaka Y, Noji S. Mito T, Mizunami M. 2016 Roles of OA1 octopamine receptor and Dop1 dopamine receptor in mediating appetitive and aversive reinforcement revealed by RNAi studies. Sci. Rep. 6, 29696.

[9] Awata H, Watanabe T, Hamanaka Y, Mito T, Noji, S., Mizunami M. 2015 Knockout crickets for the study of learning and memory: Dopamine receptor Dop1 mediates aversive but not appetitive reinforcement in crickets. Sci. Rep. 5, 15885.

[10] Mizunami M, Unoki S, Mori Y, Hirashima D, Hatano A, Matsumoto Y. 2009 Roles of octopaminergic and dopaminergic neurons in appetitive and aversive memory recall in an insect. BMC Biol. 7, 46.

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