- Alan Tudyk as House of Tomorrow
- Allison Janney as Glittering Goldie
- April Winchester as Clarabelle Cow
- Ariel Winter as Roxie Squirrel
- Audrey Wasilewski as Ortensia
- Avalon Robbins as Millie Mouse and Melody Mouse
- Benjamin Diskin as Junior Bear
- Bill Farmer as Goofy, Pluto, Fifi the Peke, Pluto's puppies, Horace Horsecollar, Practical Pig, and Penguini the Magnificent
- Billy West as Elemer Fudd
- Bob Bergen as Tweety Bird, Gabby Goat, and Clyde Bunny
- Bret Iwan as Mickey Mouse, Count Mickula, and Photo-Negative Mickey
- Brian Hull as Willie the Giant
- Candi Milo as Granny, Sweetie Bird, and Montana Max
- Carlos Alazraqui as Panchito Pistoles
- Catherine Tate as Magica De Spell
- Chris Diamantopoulos as Morty Fieldmouse, Ferdie Fieldmouse, and Steamboat Willie
- Chris Edgerly as Gyro Gearloose
- Clancy Brown as Big Bad Wolf
- Corey Burton as Ludwig von Drake, Zipper, Yen Sid, and Chernabog
- Cree Summers as Elmyra Duff and Mary Melody
- Dave Gasman as Beagle Boys
- David Errigo Jr. as Hamton J. Pig and Plucky Duck
- David Tennant as Scrooge McDuck
- Daran Norris as Wile E. Coyote
- Dee Bradley Baker as Gus Goose and Toro the Bull
- Eric Bauza as Bugs Bunny, Daffy Duck, Porky Pig, Foghorn Leghorn, Elmer Fudd, Roadrunner, José Carioca, Marvin the Martian, Buster Bunny, Tom Cat, Jerry Mouse, Egghead, Hubie & Bertie, Big Chungus, and Calamity Coyote
- Frank Welker as Oswald the Lucky Rabbit, Figaro, Road Runner, Bookworm, Bosko the Ink-Kid, Ralph the Security Guard, Sam Sheepdog, and Little Beeper
- Fred Tatasciore as Humphrey the Bear and Gossamer
- Gabriel Iglesias as Speedy Gonzales
- Giancarlo Esposito as Phantom Blot
- Grey DeLisle as Roxanne and Harietta Bear
- Harvey Guillen as Funny the Funhouse
- Hugh Davidson as Slowpoke Rodriguez
- Jason Mantzoukas as Huckster McBackstabber
- Jason Marsden as Max Goof
- Jeff Bennett as Monterey Jack and J. Audubon Woodlore
- Jeff Bergman as Sylvester the Cat, Yosemite Sam, Ralph Wolf, and Droopy McPoodle
- Jeffrey Ross as Lonesome Ghost
- Jennifer Esposito as Tina Russo
- Jess Harnell as Wakko Warner, Pete Puma, and Tosh
- Jessie Flower as Minima De Spell
- Jim Cummings as Tasmanian Devil, Tasmanian She-Devil, Pegleg Pete, Darkwing Duck / Drake Mallard, Negaduck / Jim Starling, Julius (Dr. Frankenollie's monster), Beaky Buzzard, Uncle Mortimer, and Blacque Jacque Shellacque
- Joe Hanna as Marcus Mouse
- Joe Ochman as Jiminy Cricket
- John DiMaggio as Hugo the Abominable Snowman
- Julie Brown as Minerva Mink
- Kaitlyn Robrock as Minnie Mouse, Clara Cluck, Madeline Mouse, Fifer Pig, and Fiddler Pig
- Kath Soucie as Felicity Fieldmouse, Fifi La Fume, Honey Bunny, Sniffles, and Margie Mouse
- Keith Ferguson as Flintheart Glomgold
- Kellie Martin as Mona
- Kelsey Grammer as Dr. Frankenollie and Frank Fieldmouse
- Kimiko Glenn as Lena De Spell
- Kristen Wiig as Lola Bunny
- Lara Jill Miller as Petunia Pig
- Martin Freeman as Poe de Spell
- Maurice LaMarche as Mortimer Mouse, Pepe Le Pew, The Brain, Henry Bear, and Dizzy Devil
- Natalie Palamides as Shirley the Loon and Furrball
- Nika Futterman as Cuckoo-Loca
- Paige Brewster as Della Duck
- Paul Rudish as Culpepper Mouse
- Rob Paulsen as Yakko Warner, Dr. Scratchansniff, Sweetie Bird's monster form, Mac, Steelbeak, Toodles, Pinky, and Concord Condor
- Sherri Stoner as Slappy Squirrel
- Spike Brandt as Spike the Bulldog and Tyke the Bulldog
- Tony Anselmo as Donald Duck, Donna Duck, Huey Duck, Dewey Duck, Louie Duck, and Phooey Duck
- Tress MacNeille as Daisy Duck, Dot Warner, Chip 'n' Dale, Clarice, Penelope Pussycat, Heloise Nerz, Babs Bunny, Witch Hazel, Honey the Ink-Kid, and Gadget Hackwrench
- Toks Olagundoye as Mrs. Beakley
- Tom Kenny as Mad Doctor and Fethry Duck
- "Weird Al" Yankovic as Abner "Whitewater" Duck
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