Re-oxygenation after anoxia induces brain cell death and memory loss in the anoxia-tolerant crucian carp

J Exp Biol. 2017 Nov 1;220(Pt 21):3883-3895. doi: 10.1242/jeb.165118.


Crucian carp (Carassius carassius) survive without oxygen for several months, but it is unknown whether they are able to protect themselves from cell death normally caused by the absence, and particularly return, of oxygen. Here, we quantified cell death in brain tissue from crucian carp exposed to anoxia and re-oxygenation using the terminal deoxy-nucleotidyl transferase dUTP nick-end labelling (TUNEL) assay, and cell proliferation by immunohistochemical staining for proliferating cell nuclear antigen (PCNA) as well as PCNA mRNA expression. We also measured mRNA and protein expression of the apoptosis executer protease caspase 3, in laboratory fish exposed to anoxia and re-oxygenation and fish exposed to seasonal anoxia and re-oxygenation in their natural habitat over the year. Finally, a behavioural experiment was used to assess the ability to learn and remember how to navigate in a maze to find food, before and after exposure to anoxia and re-oxygenation. The number of TUNEL-positive cells in the telencephalon increased after 1 day of re-oxygenation following 7 days of anoxia, indicating increased cell death. However, there were no consistent changes in whole-brain expression of caspase 3 in either laboratory-exposed or naturally exposed fish, indicating that cell death might occur via caspase-independent pathways or necrosis. Re-oxygenated crucian carp appeared to have lost the memory of how to navigate in a maze (learnt prior to anoxia exposure), while the ability to learn remained intact. PCNA mRNA was elevated after re-oxygenation, indicating increased neurogenesis. We conclude that anoxia tolerance involves not only protection from damage but also repair after re-oxygenation.

Keywords: Caspase 3; Cell proliferation; Fish; Gene expression; Learning.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anaerobiosis
  • Animals
  • Apoptosis
  • Brain / physiology*
  • Carps / physiology*
  • Caspase 3 / genetics
  • Caspase 3 / metabolism
  • Cell Death*
  • Female
  • Fish Proteins / genetics
  • Fish Proteins / metabolism
  • In Situ Nick-End Labeling / veterinary
  • Male
  • Memory*
  • Seasons
  • Spatial Learning*


  • Fish Proteins
  • Caspase 3