Abstract
Cope's gray tree frog (Hyla chrysoscelis) can tolerate freezing of its extracellular water. While the exact mechanism of freeze tolerance is unknown, aquaporins may play a role by allowing water and solutes like glycerol to move between compartments prior to and during freezing. Previously, two aquaporins (HC1, HC2) and one glyceroporin (HC3) were identified in Hyla chrysoscelis in a tissue and temperature dependent manner. To further elucidate the role of aquaporins in freeze tolerance, three putative aquaporins (HC7, HC8 and HC0) have been identified. Degenerate PCR primers were developed using conserved amino acid regions from Xenopus tropicalis and compared to other species with high sequence homology. For the putative aquaporins, mRNA was isolated from the fat, large intestines, and the eye for HC7, HC8 and HC0 respectively. The templates were amplified using RT‐PCR and products (326 bp, 377 bp and 400 bp respectively) were examined by electrophoresis. Further experiments are underway to confirm these putative aquaporins, and to examine their tissue and temperature dependence. Supported by NSF IOB‐0517301.
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