The series, which ran from 2000 to 2002, starred Harley Quinn, Poison Ivy, Batgirl, Catwoman, Renee Montoya and Zatanna in short stories of varying length about the daily lives of the characters.

Dropping the magician Zatanna from its lineup and adding a host of new characters to the cast, including Detective Renee Montoya, each episode from this series tied into the next, forming a collective half-hour whodunit.

The storyline also dealt with issues not seen since Batman: The Animated Series, such as the corruption of the Gotham City Police Department, Poison Ivy's environmental crusade and Commissioner Gordon's wavering faith in costumed superheroes.

The final episode was dedicated to the memory of Hilary J. Bader, script writer for numerous DC Animated series, and who had died in November 7, 2002, of breast cancer.

Sayantan Gayen, writing for CBR, praised Gotham Girls for its women-led narrative, humor, and "unique flavor" within the DC Animated Universe.

While noting the dated Flash animation, Gayen highlighted the series' strong character dynamics, clever storytelling, and its evolution from lighthearted antics to darker, more ambitious themes in its final season.