Skip to main content
Dryad logo

Glucose transporter expression and regulation following a fast in the ruby-throated hummingbird, Archilochus colubris

Citation

Ali, Raafay Syed et al. (2020), Glucose transporter expression and regulation following a fast in the ruby-throated hummingbird, Archilochus colubris, Dryad, Dataset, https://doi.org/10.5061/dryad.98sf7m0h4

Abstract

Hummingbirds, subsisting almost exclusively on nectar sugar, face extreme challenges to blood sugar regulation. The capacity for transmembrane sugar transport is mediated by the activity of facilitative glucose transporters (GLUTs) and their localisation to the plasma membrane (PM). In this study, we determined the relative protein abundance of GLUT1, GLUT2, GLUT3, and GLUT5 via immunoblot using custom antibodies in whole-tissue and PM fractions of flight-muscle, heart, and liver of ruby-throated hummingbirds (Archilochus colubris). GLUTs examined were detected in nearly all tissues tested. Hepatic GLUT1 was minimally present in whole-tissue and absent in PM fractions. GLUT5 was expressed in flight-muscles at levels comparable to that of their liver, consistent with hummingbird flight-muscles’ hypothesised uniquely high fructose-uptake and oxidation capacity. To assess GLUT regulation, we fed ruby-throated hummingbirds 1M sucrose ad libitum for 24 hours followed by either 1 hour of fasting or continued feeding until sampling. We measured relative GLUT abundance and concentrations of circulating sugars. Blood fructose concentration in fasted hummingbirds declined (∼5 mM to ∼0.18 mM), while fructose-transporting GLUT2 and GLUT5 abundance did not change in PM fractions. Blood glucose concentrations remained elevated in fed and fasted hummingbirds (∼30 mM), while glucose-transporting GLUT1 and GLUT3 in flight muscle and liver PM fractions, respectively, declined in fasted birds. Our results suggest that glucose uptake capacity is dynamically reduced in response to fasting, allowing for maintenance of elevated blood glucose levels, while fructose uptake capacity remains constitutively elevated promoting depletion of blood total fructose within the first hour of a fast.

Methods

This dataset is the raw normalised arbitrary intensity of Western blots of GLUT1, GLUT2, GLUT3, and GLUT5 from either whole-tissue homogenates ("ffGLUTdata_cell.csv") or plasma-membrane fractions ("ffGLUTdata_pm.csv") of the flight muscle, heart, and liver of fed or fasted ruby-throated hummingbirds. Each antibody band has been normalised to the corresponding total-protein stain (Sypro Ruby Red) using BioRad ImageLab software. "BirdID" refers to the internal numbering scheme for each unique hummingbird. "Blot" refers to unique immunoblot membranes. 

RScripts that were used process this data into mixed-effects models, estimate the model means, and plot the output are also provided ("GLUT ANALYSIS v2.R")

Data are also provided for the analysis of circulating sugars and metabolite concentrations with the R script entitled "ff2018 Metabolomics.R".

Usage Notes

Please make sure to have all R libraries loaded in the order they are presented in. Please note the the R scripts are not self-executable and are designed for use with R-Studio.

Funding

Natural Sciences and Engineering Research Council of Canada, Award: 386466

Human Frontier Science Program, Award: RGP0062/2016