Abstract
Learning is critical for survival as it provides the capacity to adapt to a changing environment. At the molecular and cellular level, learning leads to alterations within neural circuits that include synaptic rewiring, synaptic plasticity, and protein level/gene expression changes. There has been substantial progress in recent years on dissecting how learning and memory is regulated at the molecular and cellular level, including the use of compact invertebrate nervous systems as experimental models. This progress has been facilitated by the establishment of robust behavioral assays that generate a quantifiable readout of the extent to which animals learn and remember. This chapter will focus on protocols of behavioral tests for associative learning using the nematode Caenorhabditis elegans, with its unparalleled genetic tractability, compact nervous system of ~300 neurons, high level of conservation with mammalian systems, and amenability to a suite of behavioral tools and analyses. Specifically, we will provide a detailed description of the methods for two behavioral assays that model associative learning, one measuring appetitive olfactory learning and the other assaying aversive gustatory learning.
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References
Magee JC, Grienberger C (2020) Synaptic plasticity forms and functions. Annu Rev Neurosci 43:95–117. https://doi.org/10.1146/annurev-neuro-090919-022842
Byrne JH, Hawkins RD (2015) Nonassociative learning in invertebrates. Cold Spring Harb Perspect Biol 7(5). https://doi.org/10.1101/cshperspect.a021675
Hawkins RD, Byrne JH (2015) Associative learning in invertebrates. Cold Spring Harb Perspect Biol 7(5). https://doi.org/10.1101/cshperspect.a021709
Ardiel EL, Rankin CH (2010) An elegant mind: learning and memory in Caenorhabditis elegans. Learn Mem 17(4):191–201. https://doi.org/10.1101/lm.960510
Rahmani A, Chew YL (2021) Investigating the molecular mechanisms of learning and memory using Caenorhabditis elegans. J Neurochem 159(3):417–451. https://doi.org/10.1111/jnc.15510
Corsi AK, Wightman B, Chalfie M (2015) A transparent window into biology: a primer on Caenorhabditis elegans. Genetics 200(2):387–407. https://doi.org/10.1534/genetics.115.176099
Nance J, Frokjaer-Jensen C (2019) The Caenorhabditis elegans transgenic toolbox. Genetics 212(4):959–990. https://doi.org/10.1534/genetics.119.301506
Stiernagle T (2006) Maintenance of C. elegans. In: Community TCeR, editor. WormBook: WormBook
Fang-Yen C, Alkema MJ, Samuel AD (2015) Illuminating neural circuits and behaviour in Caenorhabditis elegans with optogenetics. Philos Trans R Soc Lond Ser B Biol Sci 370(1677):20140212. https://doi.org/10.1098/rstb.2014.0212
Kerr RA (2006) Imaging the activity of neurons and muscles. WormBook 1–13. https://doi.org/10.1895/wormbook.1.113.1
Saeki S, Yamamoto M, Iino Y (2001) Plasticity of chemotaxis revealed by paired presentation of a chemoattractant and starvation in the nematode Caenorhabditis elegans. J Exp Biol 204(Pt 10):1757–1764. https://doi.org/10.1242/jeb.204.10.1757
Torayama I, Ishihara T, Katsura I (2007) Caenorhabditis elegans integrates the signals of butanone and food to enhance chemotaxis to butanone. J Neurosci 27(4):741–750. https://doi.org/10.1523/JNEUROSCI.4312-06.2007
Stein GM, Murphy CT (2014) C. elegans positive olfactory associative memory is a molecularly conserved behavioral paradigm. Neurobiol Learn Mem 115:86–94. https://doi.org/10.1016/j.nlm.2014.07.011
Fadda M, De Fruyt N, Borghgraef C, Watteyne J, Peymen K, Vandewyer E et al (2020) NPY/NPF-related neuropeptide FLP-34 signals from serotonergic neurons to modulate aversive olfactory learning in Caenorhabditis elegans. J Neurosci 40(31):6018–6034. https://doi.org/10.1523/JNEUROSCI.2674-19.2020
Kauffman A, Parsons L, Stein G, Wills A, Kaletsky R, Murphy C. C. elegans positive butanone learning, short-term, and long-term associative memory assays. J Vis Exp. 2011(49). https://doi.org/10.3791/2490
Bargmann CI (2006) Chemosensation in C. elegans. WormBook 1–29. https://doi.org/10.1895/wormbook.1.123.1
Sagasti A, Hisamoto N, Hyodo J, Tanaka-Hino M, Matsumoto K, Bargmann CI (2001) The CaMKII UNC-43 activates the MAPKKK NSY-1 to execute a lateral signaling decision required for asymmetric olfactory neuron fates. Cell 105(2):221–232. https://doi.org/10.1016/s0092-8674(01)00313-0
Wes PD, Bargmann CI (2001) C. elegans odour discrimination requires asymmetric diversity in olfactory neurons. Nature 410(6829):698–701. https://doi.org/10.1038/35070581
Bargmann CI, Hartwieg E, Horvitz HR (1993) Odorant-selective genes and neurons mediate olfaction in C. elegans. Cell 74(3):515–527. https://doi.org/10.1016/0092-8674(93)80053-h
Tsunozaki M, Chalasani SH, Bargmann CI (2008) A behavioral switch: cGMP and PKC signaling in olfactory neurons reverses odor preference in C. elegans. Neuron 59(6):959–971. https://doi.org/10.1016/j.neuron.2008.07.038
Kodama E, Jurado P (2007) Butanone: the memory of a scent. J Neurosci 27(20):5267–5268. https://doi.org/10.1523/JNEUROSCI.1322-07.2007
Nagashima T, Iino Y, Tomioka M (2019) DAF-16/FOXO promotes taste avoidance learning independently of axonal insulin-like signaling. PLoS Genet 15(7):e1008297. https://doi.org/10.1371/journal.pgen.1008297
Baugh LR, Hu PJ (2020) Starvation responses throughout the Caenorhabditis elegans life cycle. Genetics 216(4):837–878. https://doi.org/10.1534/genetics.120.303565
Lim JP, Fehlauer H, Das A, Saro G, Glauser DA, Brunet A et al (2018) Loss of CaMKI function disrupts salt aversive learning in C. Elegans. J Neurosci 38(27):6114–6129. https://doi.org/10.1523/JNEUROSCI.1611-17.2018
Suzuki H, Thiele TR, Faumont S, Ezcurra M, Lockery SR, Schafer WR (2008) Functional asymmetry in Caenorhabditis elegans taste neurons and its computational role in chemotaxis. Nature 454(7200):114–117. https://doi.org/10.1038/nature06927
White JG, Southgate E, Thomson JN, Brenner S (1986) The structure of the nervous system of the nematode Caenorhabditis elegans. Philos Trans R Soc Lond Ser B Biol Sci 314(1165):1–340
Tomioka M, Adachi T, Suzuki H, Kunitomo H, Schafer WR, Iino Y (2006) The insulin/PI 3-kinase pathway regulates salt chemotaxis learning in Caenorhabditis elegans. Neuron 51(5):613–625. https://doi.org/10.1016/j.neuron.2006.07.024
Ohno H, Sakai N, Adachi T, Iino Y (2017) Dynamics of presynaptic diacylglycerol in a sensory neuron encode differences between past and current stimulus intensity. Cell Rep 20(10):2294–2303. https://doi.org/10.1016/j.celrep.2017.08.038
Satoh Y, Sato H, Kunitomo H, Fei X, Hashimoto K, Iino Y (2014) Regulation of experience-dependent bidirectional chemotaxis by a neural circuit switch in Caenorhabditis elegans. J Neurosci 34(47):15631–15637. https://doi.org/10.1523/JNEUROSCI.1757-14.2014
Gray JM, Hill JJ, Bargmann CI (2005) A circuit for navigation in Caenorhabditis elegans. Proc Natl Acad Sci U S A 102(9):3184–3191. https://doi.org/10.1073/pnas.0409009101
Ogg S, Ruvkun G (1998) The C. elegans PTEN homolog, DAF-18, acts in the insulin receptor-like metabolic signaling pathway. Mol Cell 2(6):887–893. https://doi.org/10.1016/s1097-2765(00)80303-2
Lackner MR, Nurrish SJ, Kaplan JM (1999) Facilitation of synaptic transmission by EGL-30 Gqalpha and EGL-8 PLCbeta: DAG binding to UNC-13 is required to stimulate acetylcholine release. Neuron 24(2):335–346. https://doi.org/10.1016/s0896-6273(00)80848-x
Brenner S (1974) The genetics of Caenorhabditis elegans. Genetics 77(1):71–94. https://doi.org/10.1002/cbic.200300625
Acknowledgments
The authors gratefully acknowledge the Caenorhabditis Genetics Center, which is supported by the National Institutes of Health (P40 OD010440), for providing strains used in data presented here. We also acknowledge our funding sources: Y.L.C is funded by the National Health and Medical Research Council (NHMRC) (GNT1173448), the Rebecca L Cooper Medical Research Foundation (PG2020652), the Flinders University Impact Seed Funding Grant (2022), and the Flinders Foundation (Mary Overton Senior Research Fellowship). A.R is funded by a PhD scholarship supported through the Australian Graduate Research Training Program. A.L.M. is supported by a Flinders University Research Scholarship (Flinders University).
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Rahmani, A., McMillen, A., Allen, E., Minervini, C., Chew, Y.L. (2024). Behavioral Tests for Associative Learning in Caenorhabditis elegans. In: Dworkin, S. (eds) Neurobiology. Methods in Molecular Biology, vol 2746. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3585-8_2
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DOI: https://doi.org/10.1007/978-1-0716-3585-8_2
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