Saddleback Journal of Biology - Saddleback College

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Spring 2010 Biology 3B Paper investigation were acclimated to temperatures averaging about 20 ˚C ± 5 ˚C; therefore, investigators hypothesized that Hyla regilla collected from more temperate climates would not exhibit significant increases in plasma glucose associated with cold temperature. Blood glucose levels significantly increased as the temperature decreased (Figure 2). The mean glucose level of the control group after protocol was 31.70 ± 2.29 mg • dL -1 (±SEM, n=10), and the mean glucose level of the near freezing group was 50.20 ± 3.10 mg • dL -1 (±SEM, n=10). The significant findings in this experiment were unexpected and did not support the investigators hypothesis. The increase in blood glucose levels suggests that glucose, a cryoprotectant, accumulates in order to prevent cellular damage by stabilizing proteins and maintaining membrane structure. Glucose may also limit intracellular dehydration via osmotic and water-binding effects. As a result, the investigators were interested in furthering their research to determine why Hyla regilla, living in Southern California, were able to survive exposure to near freezing temperatures when they had not previously been acclimated to these conditions. Therefore, the questions is, why is this adaptation observed in species that are not subjected to colder temperatures? Investigators wanted to determine if Hyla regilla increased their blood glucose levels as a response to environmental changes that may cause great stress on their body. They wanted to expose the Hyla regilla to a high saline environment, a different environmental variable that the species had not been acclimated to, in order to determine if the frogs would increase blood glucose levels. The investigators hypothesized that the blood glucose levels of Hyla regilla would increase as a response to an environment of high salinity as a protective mechanism against cellular damage. Twelve Hyla regilla were collected at a freshwater pond in Irvine, California on April 7, 2010. One control group (freshwater) and three experiment groups, consisting of three frogs each, were used in this experiment. The frogs were placed in individual petri dishes containing either water from the freshwater pond or dilutions of normal saline (50 mM NaCl, 100 mM NaCl, and 150 mM NaCl). The blood glucose levels of all the frogs were recorded before experimentation, after three hours and after 48 hours of continuous exposure to either pond water (control) or saline solutions (experimental groups). Multiple one-tailed, paired t-tests (p 0.05) were used to analyze the data. P values were analyzed to determine if the changes in blood glucose levels were significant. Investigators looked at p values that were obtained by comparing the initial blood glucose levels of each group and the blood glucose levels after three hours of continuous exposure to the solutions. In addition, p values were calculated between the initial blood glucose levels and the blood glucose levels after 48 hours of continuous exposure. There was no significant increase in the blood glucose levels in any group. Figure 3 illustrates the mean blood glucose levels of the control and experimental groups. Although unexpected, the insignificant results found in the study of exposure to a saline environment may actually provide more support towards the preliminary study. The evidence of the initial study suggested that frogs in warmer climates possess the same freeze adaptation as frogs in cold climates. However, the investigators did not put the frogs in any other stressful situations and this left the investigators’ speculation vulnerable to other arguments. The new data, showing the frogs subjected to a different environmental change with no significant increase in glucose, may reinforce the researchers’ results that suggest that a widespread distribution of the freeze tolerance adaptation exists in this species. 43 Saddleback Journal of Biology Spring 2010
Spring 2010 Biology 3B Paper Blood Glucose Concentrations (mg/dL) 50 40 30 20 10 0 Pond Water 50mM NaCl 100mM NaCl 150mM NaCl Initial Blood Glucose Blood Glucose after 3 hrs Blood Glucose after 48 hrs Control and Experimental Groups Figure 3. The mean blood glucose levels of the control group (pond water) and the experimental groups (50mM NaCl, 100mM NaCl, 150mM NaCl) before, after three hours, and after 48 hours of continuos exposure to different solutions. Literature Cited Costanzo et al. (1993). Glucose Concentration Regulates Freeze Tolerance in the Wood Frog Rana sylvatica. Physiological and Biochemical Zoology, 76(3), 331-338. Croes, S.A. & Thomas, R.E. (2000). Freeze Tolerance and Cryoprotectant Synthesis of the Pacific Tree Frog, Hyla regilla. Copeia, 2000(3), 863-868. Cunningham, J.D. & Mullally, D.P. (1956). Thermal Factors in the Ecology of the Pacific Treefrog. Herpetologica, 12(1), 68-79. Storey, K.B. & Storey, J.M. (1987). Persistence of Freeze Tolerance in Terrestrially Hibernating Frogs after Spring Emergence. Copeia, 1987(3), 720-726. Voituron, et al. (2002). To Freeze or Not to Freeze? An Evolutionary Perspective on the Cold- Hardiness Strategies of Overwintering Ectotherms. The American Naturalist, 160(2), 255-270. 44 Saddleback Journal of Biology Spring 2010
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Page 5 and 6: Author(s) Title Page Angela C. Park
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