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. 2010 Jun;7(2):205-13.
doi: 10.1089/zeb.2009.0640.

Blood sugar measurement in zebrafish reveals dynamics of glucose homeostasis

Stefani C Eames  1 , Louis H Philipson, Victoria E Prince, Mary D Kinkel
Affiliations

Blood sugar measurement in zebrafish reveals dynamics of glucose homeostasis

Stefani C Eames et al. Zebrafish. 2010 Jun.

Abstract

The adult zebrafish has the potential to become an important model for diabetes-related research. To realize this potential, small-scale methods for analyzing pancreas function are required. The measurement of blood glucose level is a commonly used method for assessing beta-cell function, but the small size of the zebrafish presents challenges both for collecting blood samples and for measuring glucose. We have developed methods for collecting microsamples of whole blood and plasma for the measurement of hematocrit and blood glucose. We demonstrate that two hand-held glucose meters designed for use by human diabetics return valid results with zebrafish blood. Additionally, we present methods for fasting and for performing postprandial glucose and intraperitoneal glucose tolerance tests. We find that the dynamics of zebrafish blood glucose homeostasis are consistent with patterns reported for other omnivorous teleost fish.

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Figures

FIG. 1.
FIG. 1.
Anesthetic comparison. Treatment with MS-222 causes increased variability in blood glucose relative to treatment with cold water. Fish were fasted for 2 h before treatment. Glucose was measured with Meter B. MS-222: mean and standard deviation, 72.91 ± 39.21 mg/dL; coefficient of variation (CV) = 54%; n = 11. Hypothermia: mean and standard deviation, 60.18 ± 17.58 mg/dL; CV = 29%; n = 11.
FIG. 2.
FIG. 2.
Glucose meter comparisons. (A) Two different glucose meters give different results. A blood sample was collected from individual fish, and each sample (n = 31) was measured immediately with both meters. A broad range of glucose values was compared by measuring fish that had been recently fed (beginning at 15 min postprandial) as well as fish that had been fasted for up to 8 days. (B) Blood glucose measured with pyrroloquinoline quinone glucose dehydrogenase (Meters B–D) using different sample sizes, compared with glucose oxidase (Meter A). Test strip sample volumes: Meter A, 1.0 μL; Meter B, 0.3 μL; Meter C, 0.6 μL; Meter D, 1.5 μL. Samples (n = 10) from individual fish were measured once with each meter. Order of meter use was randomized. Tukey's multiple comparison posttest revealed that Meter B versus Meter C was not statistically significant (p > 0.05). All other comparisons were significant at p < 0.001, except Meter D versus Meter A was significant at p < 0.05. Data are mean and standard deviation.
FIG. 3.
FIG. 3.
Blood glucose response to fasting and feeding. (A) Fasting blood glucose at 24 h: CV = 30%, n = 35; 48 h: CV = 27%, n = 11; 72 h: CV = 31%, n = 25; 96 h: CV = 21%, n = 15. Kruskal–Wallis test with Dunn's posttest showed the following: 24 h versus 48 h, p > 0.05; 24 h versus 72 h, p < 0.01; 72 h versus 96 h, p > 0.05. Data are mean and standard deviation. (B) Postprandial blood glucose. Fish were fasted for 4 days, and then fed. Sample size is a minimum of 10 fish per time point. All fed groups are significantly different from fasted, p < 0.01; glucose at 30 min versus 15 min, p < 0.05; glucose at 45 min cannot be statistically discriminated from glucose at 15 or 30 min, p > 0.05. ANOVA with Tukey's posttest. Data are mean and standard deviation. Glucose was measured with Meter B for both experiments.
FIG. 4.
FIG. 4.
Glucose tolerance test. Fish were injected with 1 mg/g glucose solution or vehicle. Glucose was measured with Meter A. Samples sizes: glucose-injected, n = 4–6; vehicle-injected, n = 3–7; noninjected, n = 10. ANOVA with Bonferroni's posttest showed no significant difference between vehicle-injected and noninjected controls for any time point. All glucose-injected were significantly different from vehicle-injected at p < 0.001, except that for 360 min postinjection there was no significant difference (p > 0.05). Data are mean and standard deviation.
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