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Cope's gray treefrog (Dryophytes chrysoscelis) is one of few freeze-tolerant frogs that mobilize glycerol as a cryoprotectant, yet cold and freezing-induced accumulation of this and other osmolytes has received little attention in this species. This study investigated the development of freeze tolerance in D. chrysoscelis, analyzing the response of the cryoprotectant system to cold acclimation, freezing, and thawing. Glycerol production was low and unresponsive to acclimation temperature, or duration of acclimation to 5 °C, except for one cold-acclimated frog that presented elevated glycerol in plasma, liver, and skeletal muscle. Curiously, glycerol concentration in skeletal muscle was higher than that of plasma and liver, in both warm- and cold-acclimated frogs, suggesting glycerol synthesis in muscle. Urea concentration in plasma doubled in response to cold acclimation but did not change during freezing. Freezing induced hepatic glycogen catabolism and an increase in glycerol and glucose in several tissues, although the mobilization dynamics differed between these cryoprotectants, possibly as a result of different transport mechanisms. Although hepatic glucose mobilization was of considerable magnitude, glucose accumulation in peripheral tissues was low and was surpassed by that of glycerol and urea. The muscle production of glycerol and the cold-induced accumulation of urea imply a role for skeletal muscle metabolism in the mobilization of cryoprotective solutes in D. chrysoscelis. The cryoprotectant system of D. chrysoscelis is complex, highly variable, and unique, with glycerol, glucose, and likely urea serving as cryoprotectants.

Cope's gray treefrogs, Hyla chrysoscelis, accumulate glycerol during the period of cold acclimation that leads to the development of freeze tolerance. Glycerol must cross cell membranes in numerous processes during this time, including exit from hepatocytes where glycerol is synthesized and entry into other tissues, where glycerol is cryoprotective. Thus, we hypothesized that erythrocytes from H. chrysoscelis would be permeable to glycerol and that that permeability would be up-regulated during cold acclimation. Further, we hypothesized that glycerol permeability would be associated with the expression of aquaporins, particularly those from the glyceroporin sub-family. Erythrocytes from warm-acclimated treefrogs had high glycerol permeability at 20°C, as assessed by the time required for osmotic lysis following suspension in 0.2 M glycerol. That osmotic lysis, as well as uptake of radio-labeled glycerol, was inhibited by 0.3 mM HgCl(3). Permeability assessed via osmotic lysis was markedly reduced at 5°C. These properties were similar in animals deriving from northern (Ohio) and southern (Alabama) populations, although suggestive (through statistical interactions) of greater glycerol permeability in northern animals. Erythrocytes expressed mRNA and protein for a previously described glyceroporin, HC-3. In cold-acclimated animals, HC-3 protein expression was up-regulated, but we could not detect a concomitant enhancement of glycerol permeability.

Renal function and osmoregulation were studied in bobwhite quail (Colinus virginianus) raised with unrestricted water (chronically unrestricted group) or restricted water (chronically restricted group). There was no difference in urine concentrating ability between adult and juvenile (3.5 or 7.5 week-old) quail. A filtration marker (polyethylene glycol) was infused into adult quail via osmotic minipumps and responses to the following regimens studied: ad libitum water intake, short-term (4-day) water restriction, and acute (1-day) dehydration (withdrawal of all drinking water). Chronically restricted quail had higher urine-to-plasma ratios of polyethylene glycol and lower urine flow rates during short-term restriction. A greater proportion of the reduction in urine flow rate during dehydration was attributable to enhanced tubular reabsorption, rather than reduced rates of filtration, in chronically restricted than in chronically unrestricted birds. Chronically restricted birds also had higher maximum urine-to-plasma ratios of polyethylene glycol (but not higher urine osmolality). These differences occurred in the face of arginine vasotocin concentrations that were not different in the two groups of birds (approximately 15 pg.ml-1 during hydration, and 45 pg.ml-1 during water restriction or dehydration). These observations suggest that chronically restricted quail have an enhanced responsiveness of tubular reabsorption to dehydration, a finding consistent with previous observations of tubule hypertrophy and hyperplasia in these birds (Goldstein and Ellis 1991). Despite this, no difference was found in medullary cAMP levels, either basal or arginine vasotocin- or forskolin-stimulated, in the two groups.(ABSTRACT TRUNCATED AT 250 WORDS)

The effects of dehydration and hemorrhage on plasma ionic, osmotic, and antidiuretic hormone (arginine vasotocin) concentrations and of hemorrhage on salt gland secretion and glomerular filtration rate were evaluated in glaucous-winged gulls, Larus glaucescens. Dehydration for 24 h did not affect plasma ionic, osmotic or arginine vasotocin concentrations; 72 h dehydration significantly elevated plasma osmolality, plasma sodium and chloride concentrations, and plasma arginine vasotocin concentration, but did not affect plasma potassium concentration. Constant infusion of 0.8 mol.1-1 NaCl increased plasma arginine vasotocin concentration and produced salt gland secretion in seven gulls; four secreted well, while three secreted less well. Removal of 20% blood volume during saline infusion immediately reduced (P<0.001) salt gland secretion rate in all gulls. After bleeding, good secretors maintained glomerular filtration rate and urine flow rate; the poorer secretors increased glomerular filtration rate and became diuretic. Blood replacement returned salt gland secretion rate to the prebleeding level (P<0.05) without affecting salt gland secretions sodium concentration in gulls which secreted well, but did not restimulate salt gland secretion in gulls which secreted poorly. Reinfusion of blood had no effect on glomerular filtration rate. Bleeding and blood replacement did not affect plasma arginine vasotocin concentration.

The contributions of the kidneys, the small intestine and the lower intestine (rectum plus cloaca) to water conservation during dehydration in unanaesthetized, unrestrained house sparrows (Passer domesticus) were assessed. Thirty hours of acute dehydration resulted in a 12% loss in body mass and a significant increase in plasma osmolality. Glomerular filtration rate declined by 55%, from 7.7 to 3.5 ml/h, and urine flow rate delined by more than 80%, from 0.2 to 0.03 ml/h. These changes are likely attributable to a large increase in plasma levels of arginine vasotocin during dehydration, from less than 26 pg/ml in hydrated birds to greater than 200 pg/ml after 30 h dehydration. Flow of water from the ileum to the lower intestine was reduced during dehydration, primarily because of a reduced flow of dry matter (with no significant reduction in water content). The rate of water loss in the excreta declined from 0.2 ml/h in hydrated birds to 0.04 ml/h in dehydrated birds. The rate of water reabsorption in the lower intestine (equal to the rate of water loss in the excreta minus the combined rates of inflow into the lower intestine from the urine and the ileal contents) slightly exceeded the rate of water flow from the ileum in both hydrated and dehydrated birds. We suggest that much of the water reabsorbed in the lower intestine of hydrated birds derives from the urine, but that primarily water from ileal contents is reabsorbed in dehydrated birds. That is, urine undergoes significant post-renal modification in hydrated but not dehydrated house sparrows.