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First published online January 19, 2006
Journal of Experimental Biology 209, 399-406 (2006)
Published by The Company of Biologists 2006
doi: 10.1242/jeb.02001
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Rapid cold-hardening increases the freezing tolerance of the Antarctic midge Belgica antarctica

Richard E. Lee, Jr1,*, Michael A. Elnitsky1, Joseph P. Rinehart2, Scott A. L. Hayward2,3, Luke H. Sandro1 and David L. Denlinger2

1 Department of Zoology, Miami University, Oxford OH 45056, USA
2 Department of Entomology, Ohio State University, 318 W. 12th Avenue, Columbus, OH 43210, USA
3 School of Biological Sciences, Liverpool University, Crown Street, Liverpool, L69 7ZB, UK


Figure 1
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  		Fig. 1. Comparison of the cold tolerance of summer-acclimatized larvae and adults and cold-acclimated larvae of the Antarctic midge Belgica antarctica. Three replicates of 10 individuals were transferred from 4°C to the specified temperatures for 24 h. Survival was assessed following a 24 h recovery at 4°C, and all individuals that displayed at least some movement were deemed to have survived. Values are means ± s.e.m.

 

Figure 2
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  		Fig. 2. Effect of rapid cold-hardening on the freeze tolerance of (A) summer-acclimatized larvae and (B) adults of the Antarctic midge, Belgica antarctica. Cultures were maintained at 4°C prior to exposure to the indicated temperatures. * denotes a significant (P<0.05) difference in survival between groups directly exposed to -10°C and those that were first held at -5°C for 1 h prior to the -10°C exposure. Values are means ± s.e.m. based on six replicates of 10 individuals.

 

Figure 3
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  		Fig. 3. Effect of rapid cold-hardening on the freeze tolerance of cold-acclimated larvae of the Antarctic midge Belgica antarctica. * denotes a significant (P<0.05) difference in survival between groups directly exposed to -15 or -20°C and those that were first held at -5°C for 1 h prior to the lower temperature exposure. Values are means ± s.e.m. based on six replicates of 10 individuals.

 

Figure 4
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  		Fig. 4. The effect of rapid cold-hardening (RCH) on cellular survival of various tissues from cold-acclimated larvae of Belgica antarctica. Green indicates living cells, and red indicates dead cells. Scale bar, 300 µm for rapidly cold-hardened gut and Malpighian tubules, and Malpighian tubules frozen with no prior treatment; 150 µm for all other tissues.

 




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