Presentation Title

Comparative Studies of Myoglobin Regulation in Whales, Dolphins, and Porpoises

Start Date

November 2016

End Date

November 2016

Location

HUB 302-#111

Type of Presentation

Poster

Abstract

Myoglobin is an intracellular oxygen-carrying molecule, important for maintaining muscle function under conditions of oxygen stress. Muscle tissues from cetaceans - whales, dolphins, and porpoises – contain elevated levels of myoglobin, compared to their terrestrial relatives (cows, horses, and pigs). This is presumably due to their aquatic lifestyle, high levels of exertion, and need to dive for extended periods of time. We wish to compare the activity of cetacean myoglobin promoters to those of their terrestrial relatives to test the possibility of the occurrence of regulatory evolution in their promoter regions. This could therefore account, in part, for the elevated levels of myoglobin present in cetacean muscle tissue. Toward this end, we have cloned ~700 base pairs of the promoter region from several species, both cetacean and terrestrial, into a reporter vector expressing firefly luciferase (pGL4.10). The recombinant vectors are transfected into the mouse myoblast cell line, C2C12, through the use of Polyfect (Qiagen). The transfected cells are then grown under differentiating conditions (DMEM+2% horse serum) and assayed for firefly luciferase activity. Firefly luciferase activity is normalized to a co-transfected standard expressing Renilla luciferase. We find that the porpoise (Phocoenidae) promoter has the highest activity of the species tested, followed by baleen whales (Balaenoptera acutorostrata), humpback whales (Megaptera novaeangliae), and then dolphin (Delphinus capensis). The only terrestrial species tested extensively, the domestic pig (Sus scrofa), has 1/3 to 1/2 the activity of the cetaceans. Future directions of this study will be expanded to include other species, both cetacean and terrestrial.

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Comparative Studies of Myoglobin Regulation in Whales, Dolphins, and Porpoises

HUB 302-#111

Myoglobin is an intracellular oxygen-carrying molecule, important for maintaining muscle function under conditions of oxygen stress. Muscle tissues from cetaceans - whales, dolphins, and porpoises – contain elevated levels of myoglobin, compared to their terrestrial relatives (cows, horses, and pigs). This is presumably due to their aquatic lifestyle, high levels of exertion, and need to dive for extended periods of time. We wish to compare the activity of cetacean myoglobin promoters to those of their terrestrial relatives to test the possibility of the occurrence of regulatory evolution in their promoter regions. This could therefore account, in part, for the elevated levels of myoglobin present in cetacean muscle tissue. Toward this end, we have cloned ~700 base pairs of the promoter region from several species, both cetacean and terrestrial, into a reporter vector expressing firefly luciferase (pGL4.10). The recombinant vectors are transfected into the mouse myoblast cell line, C2C12, through the use of Polyfect (Qiagen). The transfected cells are then grown under differentiating conditions (DMEM+2% horse serum) and assayed for firefly luciferase activity. Firefly luciferase activity is normalized to a co-transfected standard expressing Renilla luciferase. We find that the porpoise (Phocoenidae) promoter has the highest activity of the species tested, followed by baleen whales (Balaenoptera acutorostrata), humpback whales (Megaptera novaeangliae), and then dolphin (Delphinus capensis). The only terrestrial species tested extensively, the domestic pig (Sus scrofa), has 1/3 to 1/2 the activity of the cetaceans. Future directions of this study will be expanded to include other species, both cetacean and terrestrial.