Rob Paulsen as Yakko Warner, Pinky and Dr.Recurring segments new to the revival include Starbox and Cindy, which follows a miniature alien who is part of a fleet who want to destroy the Earth that ends up in the hands of a young girl, The Incredible Gnome in People's Mouths, centered on the titular gnome who ventures into different people's mouths to speak for them, Math-terpiece Theater, which involves dramatized math lessons taught by Dot, and Everyday Safety, a parody of 1980's safety videos featuring the Warner siblings. Episodes are composed of several shorts, with each episode consisting of segments following the adventures of Yakko, Wakko and Dot the vast majority of episodes also include a segment featuring fan-favorite characters Pinky and the Brain-two lab mice, one of whom is intelligent and wants to take over the world, while the other is dim-witted and clumsy, often messing up his friend's plans.

Animaniacs continues to focus on the adventures of the Warner siblings, Yakko, Wakko, and Dot-three inseparable, irascible kids-as they embark on further adventures after being absent from television for 22 years, bringing with them the usual wackiness and mayhem they create while adapting to the changes and life of the 21st century.

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