Abstract
Cope's gray treefrog, Hyla chrysoscelis is a freeze‐tolerant anuran which accumulates and distributes glycerol as a cryoprotectant before freezing. We hypothesize that HC‐3, an ortholog of mammalian aquaporin 3, functions as an aquaglyceroporin to facilitate transmembrane flux of water and glycerol. HC‐3 protein is in higher abundance and is preferentially localized to the plasma membrane in RBCs from cold‐acclimated treefrogs as compared to warm‐acclimated animals. The objective of this study is to determine the signal for HC‐3 membrane translocation that occurs during the cold‐acclimation period. After 48 hrs, cultured erythrocytes from H. chrysoscelis were exposed to cAMP (1 uM; 30 minutes), vasopressin (10 IU; 30 minutes), or epinephrine (1 uM; 60 minutes), isolated from the culture media, and fixed on microscope slides. Fluorescent immunocytochemistry showed enhanced HC‐3 membrane localization in cells exposed to epinephrine and cAMP as compared to controls. These data indicate HC‐3 translocation and membrane localization is enhanced in a cAMP‐dependent pathway. This research was supported by NSF Research Grant IOS‐1121457 and the UD University Honors Program.
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