 Timothy FarrellDr. Alton / Narrator | Dr. Alton / Narrator |
 Bela LugosiScientist | Scientist |
 Edward D. Wood Jr.Glen / Glenda (as Daniel Davis) | Glen / Glenda (as Daniel Davis) |
 Lyle TalbotInspector Warren | Inspector Warren |
 Dolores FullerBarbara | Barbara |
| ''Tommy' HaynesAlan / Anne | Alan / Anne |
| CCharlie CraftsJohnny (as Charles Crafts) | Johnny (as Charles Crafts) |
 Conrad BrooksBanker / Reporter / Pickup Artist / Bearded Drag (as Connie Brooks) | Banker / Reporter / Pickup Artist / Bearded Drag (as Connie Brooks) |
| HHenry BederskiMan with Hat and Receding Hairline (uncredited) | Man with Hat and Receding Hairline (uncredited) |
| CCarol DaughertyWoman in Nightmare (uncredited) | Woman in Nightmare (uncredited) |
| ''Captain' DeZitaThe Devil / Glen's Father (uncredited) | The Devil / Glen's Father (uncredited) |
| HHelen Miles(uncredited) | (uncredited) |
| SShirley SperilMiss Stevens (uncredited) | Miss Stevens (uncredited) |
 Amzie StricklandMinor Role (uncredited) | Minor Role (uncredited) |
| HHarry ThomasMan in Nightmare (uncredited) | Man in Nightmare (uncredited) |
| WWilliam C. ThompsonJudge (uncredited) | Judge (uncredited) |
| ''Mr. Walter' HadjwiecyzPatrick / Patricia (uncredited) | Patrick / Patricia (uncredited) |
| GGeorge WeissMan at Transvestite's Suicide (uncredited) | Man at Transvestite's Suicide (uncredited) |
| EEvelyn WoodSheila, Glen's Sister (uncredited) | Sheila, Glen's Sister (uncredited) |
