Laboratory breeding of the short-lived annual killifish Nothobranchius furzeri

Abstract

Turquoise killifish, Nothobranchius furzeri, have an intrinsically short life span, with a median life span of <6 months and a maximum (90%) life span of 9 months. This short life span, which is unique among vertebrates, evolved naturally and has resulted in N. furzeri becoming a widely used laboratory model species in aging research and other disciplines. Here, we describe a protocol for the maintenance and breeding of the species under laboratory conditions. We provide details for egg incubation, hatching, everyday care of juvenile and adult fish, breeding and treatment of most common diseases. Emphasis is given to the fact that the requirements of N. furzeri substantially differ from those of other fish model taxa; N. furzeri live brief lives and in nature undergo nonaquatic embryo development, with consequences for their laboratory culture.

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Figure 1: Annual fish Nothobranchius furzeri.
Figure 2: Developmental trajectories of N. furzeri in aquatic medium (egg incubation at 25 °C).
Figure 3: External appearance of fish infected by common N. furzeri pathogens.
Figure 4: Male N. furzeri infected with Glugea sp.
Figure 5: Schematic overview of the sequence and duration of different steps in the husbandry regime of N. furzeri.
Figure 6: Basic equipment needed for husbandry of N. furzeri.
Figure 7: Gross staging of N. furzeri eggs incubated in peat.
Figure 8: Setup for hatching peat-incubated N. furzeri eggs.
Figure 9: Differences between a fertilized and an unfertilized N. furzeri egg.

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Acknowledgements

Financial support for our current research on Nothobranchius comes from the Czech Science Foundation (P506/11/0112) to M.R. We thank M. Vrtílek and three anonymous referees for comments, E. Řehulková for help with illustrations, A. Dorn for providing Figure 4 and helpful comments, and R. Spence and S. White for correcting the English. All procedures described herein are in accordance with Czech legal regulations and have been approved by the ethical committee of the Institute of Vertebrate Biology, Czech Academy of Sciences.

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M.P., R.B. and M.R. developed the protocol over the last 10 years. M.R. initiated the paper. M.P. drafted the protocol. R.B. prepared photographic documentation. All authors contributed to the final text.

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Correspondence to Matej Polačik.

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Polačik, M., Blažek, R. & Reichard, M. Laboratory breeding of the short-lived annual killifish Nothobranchius furzeri. Nat Protoc 11, 1396–1413 (2016). https://doi.org/10.1038/nprot.2016.080

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