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Potential evidence of peripheral learning and memory in the arms of dwarf cuttlefish, Sepia bandensis


CREB (cAMP response element-binding) transcription factors are conserved markers of memory formation in the brain and peripheral circuits. We provide immunohistochemical evidence of CREB phosphorylation in the dwarf cuttlefish, Sepia bandensis, following the inaccessible prey (IP) memory experiment. During the IP experiment, cuttlefish are shown prey enclosed in a transparent tube, and tentacle strikes against the tube decrease over time as the cuttlefish learns the prey is inaccessible. The cues driving IP learning are unclear but may include sensory inputs from arms touching the tube. The neural activity marker, anti-phospho-CREB (anti-pCREB) was used to determine whether IP training stimulated cuttlefish arm sensory neurons. pCREB immunoreactivity occurred along the oral surface of the arms, including the suckers and epithelial folds surrounding the suckers. pCREB increased in the epithelial folds and suckers of trained cuttlefish. We found differential pCREB immunoreactivity along the distal–proximal axis of trained arms, with pCREB concentrated distally. Unequal CREB phosphorylation occurred among the 4 trained arm pairs, with arm pairs 1 and 2 containing more pCREB. The resulting patterns of pCREB in trained arms suggest that the arms obtain cues that may be salient for learning and memory of the IP experiment.

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CAMP response element-binding


Inaccessible prey


Long-term memory


Phosphate buffered saline




pCREB immunoreactivity




Short-term memory


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We would like to thank Bedore’s lab (Dr. Christine Bedore, Theresa Gunn, and Matt Levendosky) for their valuable resource sharing and advice on cuttlefish husbandry. We would also like to thank Bret Grasse for his advice on cuttlefish rearing.


This research was funded by Georgia Southern faculty research seed award and Georgia Southern graduate student research award.

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JB and VS participated in the design of this study. JB, JW and TN performed all the experiments and data analyses. JB and VS wrote and revised the manuscript.

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Correspondence to Vinoth Sittaramane.

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All the procedures in this study were conducted according to ARRIVE guidelines.

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Bowers, J., Wilson, J., Nimi, T. et al. Potential evidence of peripheral learning and memory in the arms of dwarf cuttlefish, Sepia bandensis. J Comp Physiol A 207, 575–594 (2021).

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  • Cuttlefish
  • CREB
  • Learning
  • Memory
  • Sepia