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Am J Physiol Regul Integr Comp Physiol 261: R1346-R1350, 1991;
0363-6119/91 $5.00
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AJP - Regulatory, Integrative and Comparative Physiology, Vol 261, Issue 6 1346-R1350, Copyright © 1991 by American Physiological Society

ARTICLES

Freeze-thaw injury in erythrocytes of the freeze-tolerant wood frog, Rana sylvatica

J. P. Costanzo and R. E. Lee Jr
Department of Zoology, Miami University, Oxford, Ohio 45056.

Erythrocytes from the freeze-tolerant wood frog (Rana sylvatica) were subjected to in vitro tests of freeze tolerance, cryoprotection, and osmotic fragility. The responses of cells from frogs acclimated to 4 or 15 degrees C were similar. Erythrocytes that were frozen in saline hemolyzed at -4 degrees C or lower. The addition of high concentrations (150 and 1,500 mM) of glucose or glycerol, cryoprotectants produced naturally by freeze-tolerant frogs, significantly reduced cell injury at -8 degrees C, but concentrations of 1.5 or 15 mM were ineffective. Hemolysis was reduced by 94% with 1,500 mM glycerol and by 84% with 1,500 mM glucose; thus glycerol was the more effective cryoprotectant. Mean fragility values for frog erythrocytes incubated in hypertonic and hypotonic saline were 1,938 and 49 mosM, respectively. Survival in freeze tolerance and cryoprotection experiments was comparable for erythrocytes from frogs and humans, suggesting that these cells may respond similarly to freezing-related stresses. However, the breadth of osmotic tolerance, standardized for differences in isotonicity, was greater for frog erythrocytes than for human erythrocytes. Our data suggest that erythrocytes from R. sylvatica are adequately protected by glucose under natural conditions of freezing and thawing.


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J. P. Costanzo, J. A. Mugnano, H. M. Wehrheim, and R. E. Lee Jr.
Osmotic and freezing tolerance in spermatozoa of freeze-tolerant and -intolerant frogs
Am J Physiol Regulatory Integrative Comp Physiol, September 1, 1998; 275(3): R713 - R719.
[Abstract] [Full Text] [PDF]


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