Glen ou Glenda
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Bela LugosiScientist
Edward D. Wood Jr.Glen / Glenda (as Daniel Davis)
Lyle TalbotInspector Warren
Timothy FarrellDr. Alton / Narrator
Dolores FullerBarbara- 'Tommy' HaynesAlan / Anne
- Charlie CraftsJohnny (as Charles Crafts)
Conrad BrooksBanker / Reporter / Pickup Artist / Bearded Drag (as Connie Brooks)- Henry BederskiMan with Hat and Receding Hairline (sem créditos)
- Carol DaughertyWoman in Nightmare (sem créditos)
- 'Captain' DeZitaThe Devil / Glen's Father (sem créditos)
- Helen Miles(sem créditos)
- Shirley SperilMiss Stevens (sem créditos)
Amzie StricklandMinor Role (sem créditos)- Harry ThomasMan in Nightmare (sem créditos)
- William C. ThompsonJudge (sem créditos)
- 'Mr. Walter' HadjwiecyzPatrick / Patricia (sem créditos)
- George WeissMan at Transvestite's Suicide (sem créditos)
- Evelyn WoodSheila, Glen's Sister (sem créditos)