Because the hepatic glycogen store is substantially reduced following hibernation and mating, spring frogs can accrue only modest amounts of these agents, and this difference purportedly is the cause of their reduced freeze tolerance [4-8].
Winter and spring frogs were frozen slowly and thawed gradually following a protocol that promotes survival by facilitating cryoprotective responses and presumably mimics natural freezing and thawing episodes (i.e., slow freezing followed by gradual warming).
We could not assay urea in the plasma of spring frogs that were frozen or frozen/thawed, as too little sample was available.
We examined freeze tolerance in spring frogs by subjecting them to experimental freezing and thawing as described in the preceding section, except that these frogs (n = 6 per group) were cooled to the prescribed [T.sub.b], -5[degrees]C, -7.5[degrees]C, or -10[degrees]C, over a period of 80, 130, and 180 h, respectively.
Morphometric and physiological variables were compared between winter and spring frogs using Student's 1-tests.
Spring frogs weighed ~55% more and were 14% longer than winter frogs (Table 1).
There was congruence between winter and spring frogs in several of the response variables (Table 1).
Frogs of the two groups had similar glycemic levels, but winter frogs had plasma urea levels almost 100 [micro]mol[mL.sup.-1] higher than those in spring frogs. Due in part to the abundance of this solute, winter frogs had a markedly higher (2.24-fold) plasma osmolality (Table 1).
Observations made during tissue harvesting attested that spring frogs sampled 48 h after freezing began contained substantial amounts of ice in the coelom, beneath the skin, and within the muscles.
This was also the case with spring frogs ([F.sub.2,13] = 66.9, P < 0.0001), although the pattern of change differed between the groups ([F.sub.5,29] = 59.0, P < 0.0001).