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Journal of Comparative Physiology A

, Volume 202, Issue 6, pp 399–409 | Cite as

Training Lymnaea in the presence of a predator scent results in a long-lasting ability to form enhanced long-term memory

  • Jeremy Forest
  • Hiroshi Sunada
  • Shawn Dodd
  • Ken Lukowiak
Original Paper

Abstract

Lymnaea exposed to crayfish effluent (CE) gain an enhanced ability to form long-term memory (LTM). We test the hypothesis that a single CE exposure and operant conditioning training leads to long lasting changes in the capability of snails to form LTM when tested in pond water four weeks later. We trained both juvenile and adult snails with a single 0.5 h training session in CE and show that LTM was present 24 h later. Snails trained in a similar manner in just pond water show no LTM. We then asked if such training in CE conferred enhanced memory forming capabilities on these snails four weeks later. That is, would LTM be formed in these snails four weeks later following a single 0.5 h training session in pond water? We found that both adult and juvenile snails previously trained in CE one month previously had enhanced LTM formation abilities. The injection of a DNA methylation blocker, 5-AZA, prior to training in adult snails blocked enhanced LTM formation four weeks later. Finally, this enhanced LTM forming ability was not passed on to the next generation of snails.

Keywords

Lymnaea Long-term memory Epigenetic change Predator scent 

Abbreviations

CE

Crayfish effluent

CNS

Central nervous system

CREB

cAMP response element-binding protein

F1

First generation

LTM

Long-term memory

MT

Memory test session

PW

Pond water

SEM

Standard error of the mean

TS

Training session

5-AZA

5-Aza-2′-deoxycytidine

Notes

Acknowledgments

Funded by NSERC.

Complicance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

Ethical statement

We have followed all applicable University of Calgary protocols necessary for these experiments.

References

  1. Alexander J, Covich A (1991) Predator Avoidance by the freshwater snail Physella virgata in response to the crayfish Procambarus simulans. Oecologia 87:435–442CrossRefGoogle Scholar
  2. Braun M, Lukowiak K (2011) Intermediate and long-term memory differ at the neuronal level in Lymnaea. Neurobiol Learn Mem 6:403–416CrossRefGoogle Scholar
  3. Braun M, Lukowiak KS, Karnik V, Lukowiak K (2012) Differences in neuronal activity explain differences in memory forming abilities of different populations of Lymnaea stagnalis. Neurobiol Learn Mem 97:173–182CrossRefPubMedGoogle Scholar
  4. Crowl T, Covich A (1990) Predator-induced life-history shifts in a freshwater snail. Science 247:949–951CrossRefPubMedGoogle Scholar
  5. Dalesman S, Lukowiak K (2010) Acute exposure to low environmental calcium alters respiration and locomotion of Lymnaea stagnalis. J Exp Biol 213:1471–1476CrossRefPubMedGoogle Scholar
  6. Dalesman S, Lukowiak K (2011) Social snails: the effect of social isolation on cognition is dependent on environmental context. J Exp Biol 214:4179–4185CrossRefPubMedGoogle Scholar
  7. Dalesman S, Lukowiak K (2012) How stress alters memory in ‘smart’ snails. PloS One 7(2):e32334CrossRefPubMedPubMedCentralGoogle Scholar
  8. Dalesman S, Rundle S, Lukowiak K (2011) Microgeographic variation in memory formation following operant conditioning to a novel stimulus. Anim Behav 82:311–319CrossRefGoogle Scholar
  9. Dolinoy D, Huang D, Jirtle R (2007) Maternal nutrient supplementation counteracts bisphenol A-induced DNA hypomethylation in early development. Proc Natl Acad Sci 104:13056–13061CrossRefPubMedPubMedCentralGoogle Scholar
  10. Dominguez-Salas P, Moore SE, Baker MS, Bergen AW, Cox SE, Dyer RA, Fulford AJ, Guan Y, Laritsky E, Silver MJ, Swan GE, Zeisel SH, Innis SM, Waterland RA, Prentice AM, Hennig BJ (2014) Maternal nutrition at conception modulates DNA methylation of human metastable epialleles. Nat Comm 5:3746. doi: 10.1038/ncomms4746 CrossRefGoogle Scholar
  11. Gapp K, Jawaid A, Sarkies P, Bohacek J, Pelczar P, Prados J, Farinelli L, Miska E, Mansuy IM (2014) Implication of sperm RNAs in transgenerational inheritance of the effects of early trauma in mice. Nat Neurosci 17:667–669CrossRefPubMedPubMedCentralGoogle Scholar
  12. Giese KP, Mizuno K (2013) The roles of protein kinases in learning and memory. Learn Mem 20:540–552CrossRefPubMedGoogle Scholar
  13. Griffith J, Mahler H (1969) A ticketing of memory. Nature 223:580–582CrossRefPubMedGoogle Scholar
  14. Guan Z, Giustetto M, Lomvardas S, Kim J-H, Miniaci MC, Schwartz JH, Thanos D, Kandel ER (2002) Integration of long-term-memory-related synaptic plasticity involves bidirectional regulation of gene expression and chromatin structure. Cell 111:483–493CrossRefPubMedGoogle Scholar
  15. Haney J, Lukowiak K (2001) Context learning and the effect of context on memory retrieval in Lymnaea. Learn Mem 8:35–43CrossRefPubMedPubMedCentralGoogle Scholar
  16. Holloway T, Gonzalez-Maeso J (2015) Epigenetic mechanisms of serotonin signaling. ACS Chem Neurosci 6:1099–1109CrossRefPubMedGoogle Scholar
  17. Il-Han J, Janes T, Lukowiak K (2010) The role of serotonin in the enhancement of long-term memory resulting from predator detection in Lymnaea. J Exp Biol 213:3603–3614CrossRefPubMedGoogle Scholar
  18. Inoue T, Takasaki M, Lukowiak K, Syed N (1996) Inhibition of the respiratory pattern-generating neurons by an identified whole-body withdrawal interneuron of Lymnaea stagnalis. J Exp Biol 199:1887–1898PubMedGoogle Scholar
  19. Inoue T, Haque Z, Lukowiak K, Syed N (2001) Hypoxia-induced respiratory patterned activity in Lymnaea originates at the periphery. J Neurophys 86:156–163Google Scholar
  20. Landry C, Kandel E, Rajasethupathy P (2013) New mechanisms in memory storage: piRNAs and epigenetics. Trends Neurosci 36:535–542CrossRefPubMedGoogle Scholar
  21. Lukowiak K, Ringseis E, Spencer G, Wildering W, Syed N (1996) Operant conditioning of aerial respiratory behaviour in Lymnaea stagnalis. J Exp Biol 199:683–691PubMedGoogle Scholar
  22. Lukowiak K, Cotter R, Westly J, Ringseis E, Spencer G, Syed N (1998) Long term memory of an operantly conditioned respiratory behaviour in Lymnaea stagnalis. J Exp Biol 201:877–882PubMedGoogle Scholar
  23. Lukowiak K, Adatia N, Krygier D, Syed N (2000) Operant conditioning in Lymnaea: evidence for intermediate- and long-term memory. Learn Mem 7:140–150CrossRefPubMedPubMedCentralGoogle Scholar
  24. Lukowiak K, Sangh S, Scheibenstock A, Parvez K, McComb C, Rosengger D, Varshney N, Sadamoto H( (2003a) A molluscan model system in the search for the engram. J Physiol (Paris) 97:69–76CrossRefGoogle Scholar
  25. Lukowiak K, Sangha S, McComb C, Varshney N, Rosengger D, Sadamoto H, Scheibenstock A (2003b) Associative learning and memory in Lymnaea stagnalis: how well do they remember? J Exp Biol 206:2097–2103CrossRefPubMedGoogle Scholar
  26. Lukowiak K, Martens K, Rosenegger D, Browning K, de Caigny P, Orr M (2008) The perception of stress alters adaptive behaviours in Lymnaea stagnalis. J Exp Biol 211:1747–1756CrossRefPubMedGoogle Scholar
  27. Lukowiak K, Orr M, de Caigny P, Lukowiak KS, Il-Han J, Dalesman S (2010) Ecologically relevant stressors modify long-term memory formation in a model system. Behav Brain Res 214:18–24CrossRefPubMedGoogle Scholar
  28. Lukowiak K, Heckler B, Bennett T, Schriner E, Wyrick K, Jewett C, Todd R, Sorg B (2014) Enhanced memory persistence is blocked by DNA methyltransferase inhibitor in the snail Lymnaea stagnalis. J Exp Biol 217:2920–2929CrossRefPubMedGoogle Scholar
  29. McComb C, Meems R, Syed N, Lukowiak K (2003) Electrophysiological differences in the neuronal circuit controlling aerial respiratory behaviour between juvenile and adult Lymnaea. J Neurophys 90:983–992CrossRefGoogle Scholar
  30. McComb C, Varshney N, Lukowiak K (2005) Juvenile Lymnaea ventilate, learn and remember differently than do adult Lymnaea. J Exp Biol 208:1459–1467CrossRefPubMedGoogle Scholar
  31. Orr MV, Lukowiak K (2008) Electrophysiological and behavioral evidence demonstrating that predator detection alters adaptive behaviors in the snail Lymnaea. J Neurosci 28:2726–2734CrossRefPubMedGoogle Scholar
  32. Orr MV, El-Bekai M, Lui M, Watson K, Lukowiak K (2007) Predator detection in Lymnaea stagnalis. J Exp Biol 210:4150–4158CrossRefPubMedGoogle Scholar
  33. Orr MV, Hittel K, Lukowiak K (2009a) “Different strokes for different folks”: geographically isolated strains of Lymnaea stagnalis only respond to sympatric predators and have different memory forming capabilities. J Exp Biol 212:2237–2247CrossRefPubMedGoogle Scholar
  34. Orr MV, Hittel K, Lukowiak KS, Han J, Lukowiak K (2009b) Differences in LTM-forming capability between geographically different strains of Alberta Lymnaea stagnalis are maintained whether they are trained in the lab or in the wild. J Exp Biol 212:3911–3918CrossRefPubMedGoogle Scholar
  35. Orr MV, Hittel K, Lukowiak K (2010) Predator detection enables juvenile Lymnaea to form long-term memory. J Exp Biol 213:301–307CrossRefPubMedGoogle Scholar
  36. Rosenegger D, Lukowiak K (2010) The participation of NMDA receptors, PKC, and MAPK in the formation of memory following operant conditioning in Lymnaea. Mol Brain 3:24. doi: 10.1016/j.bbr.2010.05.011 CrossRefPubMedPubMedCentralGoogle Scholar
  37. Rosenegger D, Lukowiak K (2013) The participation of NMDA receptors, PKC, and MAPK in Lymnaea memory extinction. Neurobiol Learn Mem 100:64–69CrossRefPubMedGoogle Scholar
  38. Rosenegger D, Parvez K, Lukowiak K (2008) Enhancing memory formation by altering protein phosphorylation balance. Neurobiol Learn Mem 90:544–552CrossRefPubMedGoogle Scholar
  39. Rosenegger D, Wright C, Lukowiak K (2010) A quantative proteomic analysis of long-term memory. Mol Brain 3:9. doi: 10.1186/1756-6606-3-9 CrossRefPubMedPubMedCentralGoogle Scholar
  40. Rundle S, Bronmark C (2001) Inter-and intraspecific trait compensation of defence mechanisms in freshwater snails. Proc R Soc Lond B 268:1463–1468CrossRefGoogle Scholar
  41. Rundle SD, Spicer JI, Coleman RA, Vosper J, Soane J (2004) Environmental calcium modifies induced defences in snails. Proc R Soc Lond B 271:S67–S70CrossRefGoogle Scholar
  42. Sadamoto H, Sato H, Kobayashi S, Murakami J, Aonuma H, Ando H, Fujito Y, Hamano K, Awaji M, Lukowiak K, Urano A, Ito E (2003) CREB in the pond snail Lymnaea stagnalis: cloning, gene expression and function in identifiable neurons of the central nervous system. J Neurobiol 58:455–466CrossRefGoogle Scholar
  43. Sangha S, Scheibenstock A, Lukowiak K (2003a) Reconsolidation of a long-term memory in Lymnaea requires new protein and RNA synthesis and the soma of RPeD1. J Neurosci 23:8034–8040PubMedGoogle Scholar
  44. Sangha S, Scheibenstock A, Morrow R, Lukowiak K (2003b) Extinction requires new RNA and protein synthesis and the soma of the cell RPeD1 in Lymnaea stagnalis. J Neurosci 23:9842–9851PubMedGoogle Scholar
  45. Scheibenstock A, Krygier D, Haque Z, Syed S, Lukowiak K (2002) The soma of RPeD1 must be present for LTM formation of associative learning in Lymnaea. J Neurophys 88:1584–1591Google Scholar
  46. Spencer G, Syed N, Lukowiak K (1999) Neural changes following operant conditioning of aerial respiratory behaviour in Lymnaea stagnalis. J Neurosci 19:1836–1843PubMedGoogle Scholar
  47. Spencer G, Kazmi M, Syed N, Lukowiak K (2002) Changes in the activity of a CPG neuron after reinforcement of an operantly conditioned behavior in Lymnaea. J Neurophys 88:1915–1923Google Scholar
  48. Sunada H, Horikoshi T, Lukowiak K, Sakakibara M (2010) Increase in intrinsic excitability of RPeD11 in memory enhancement of juvenile and adult Lymnaea by predator-induced stress. Neurobiol Learn Mem 94:269–277CrossRefPubMedGoogle Scholar
  49. Syed N, Bulloch AGM, Lukowiak K (1990) In vitro reconstruction of the respiratory central pattern generator of the mollusk Lymnaea. Science 250:282–285CrossRefPubMedGoogle Scholar
  50. Syed NI, Ridgway RL, Lukowiak K, Bulloch AGM (1992) Transplantation and functional integration of an identified respiratory interneuron in Lymnaea stagnalis. Neuron 8:767–774CrossRefPubMedGoogle Scholar
  51. Takigami S, Sunada H, Lukowiak K, Sakakibara M (2014) Protein kinase C mediates memory consolidation of taste avoidance conditioning in Lymnaea stagnalis. Neurobiol Learn Mem 111:9–18CrossRefPubMedGoogle Scholar
  52. Waterland R, Jirtle R (2003) Transposable elements: targets for early nutritional effects on epigenetic gene regulation. Mol Cell Biol 15:5293–5300CrossRefGoogle Scholar
  53. Yu N, Baek S, Kaang B (2011) DNA methylation-mediated control of learning and memory. Mol Brain 4:5–11CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Jeremy Forest
    • 1
    • 2
  • Hiroshi Sunada
    • 1
  • Shawn Dodd
    • 1
  • Ken Lukowiak
    • 1
  1. 1.Hotchkiss Brain Institute, Cumming School of Medicine, University of CalgaryCalgaryCanada
  2. 2.University Claude BernardLyonFrance

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