She looks almost identical to the modern-day Mrs. Puff: a pufferfish with short, stubby fins and blonde hair color. The only difference between her and Mrs. Puff are their hairstyle and clothing. She wears a light purple dress and a long magenta feather in her hair, clipped in with a blue barrette. Her hair is longer and curlier than Mrs. Puff's.
In the song "Dead Eye," Polene tells SpongeBuck that Dead Eye makes all the girls cry. Later, she, William, and Hopalong try to ride their carriage out of town to avoid Dead Eye. However, Dead Eye catches up to them and steals the clothes off their backs, forcing them to wear their pajamas. Polene gives up and tells SpongeBuck that they've lost because Dead Eye took everything.
SpongeBuck rallies the others together and tells them if they all team up together, they can stand up to Dead Eye Plankton and run him right out of Dead Eye Gulch for good. Polene takes part in the group huddle and agrees. SpongeBuck gets the encouragement from the others to defeat Dead Eye, but they decide to hide during the actual fight.
At the end of the episode, Polene lets out her anger and stomps on Dead Eye Plankton multiple times.
Polene also appears in the book adaptation of the episode. The books Chuckle and Cringe and Unidentified Laughing Objects also feature appearances from Polene, but her design is just used as Mrs. Puff.