| Kyle MacLachlan | Dale Cooper |
| Jane Adams | Constance Talbot |
| Joe Adler | Roger |
| Stephanie Allynne | Soccer Mom |
| Chrysta Bell | FBI Agent Tammy Preston |
| Jim Belushi | Bradley Mitchum |
| Richard Beymer | Benjamin Horne |
| John Billingsley | Doctor Ben |
| Larry Clarke | Detective T. Fusco |
| Catherine E. Coulson | Margaret Lanterman (The Log Lady)
(as Catherine Coulson) |
| Giselle DaMier | Sandie |
| Jan D'Arcy | Sylvia Horne |
| David Dastmalchian | Pit Boss Warrick |
| Eric Edelstein | Detective 'Smiley' Fusco |
| Eamon Farren | Richard Horne |
| Miguel Ferrer | FBI Agent Albert Rosenfield |
| Patrick Fischler | Duncan Todd |
| Pierce Gagnon | Sonny Jim Jones |
| Michael Horse | Deputy Chief Tommy 'Hawk' Hill |
| Caleb Landry Jones | Steven Burnett |
| David Patrick Kelly | Jerry Horne |
| Robert Knepper | Rodney Mitchum |
| David Koechner | Detective D. Fusco |
| Andrea Watrouse | Mandie
(as Andrea Leal) |
| Sheryl Lee | Laura Palmer
(archive footage) |
| Sarah Jean Long | Miriam Sullivan |
| David Lynch | FBI Deputy Director Gordon Cole |
| Greg Mills | Paul |
| Moby | Musician |
| John Pirruccello | Deputy Chad Broxford |
| Kimmy Robertson | Lucy Brennan |
| Wendy Robie | Nadine Hurley |
| Erik Rondell | Johnny Horne |
| Amanda Seyfried | Rebecca 'Becky' Burnett |
| Amy Shiels | Candie |
| Tom Sizemore | Anthony Sinclair |
| Sara Sohn | Sheena |
| Harry Dean Stanton | Carl Rodd |
| Russ Tamblyn | Dr. Lawrence Jacoby |
| Naomi Watts | Janey-E Jones |
| Christophe Zajac-Denek | Ike 'The Spike' Stadtler |
| Rebekah Del Rio | Rebekah Del Rio |
| Mick Flowers | Drummer
(uncredited) |
| Nick Launay | Keyboardist
(uncredited) |
| Grace Zabriskie | Sarah Palmer
(voice) (uncredited) |