Ivan Trojan as Myšpulín (voice)

Ondřej Gregor Brzobohatý as Piňďa (voice)

Tereza Bebarová as Fifinka (voice)

Bohdan Tůma as Bobík (voice)

Václav Postránecký as Rudolf II. (voice)

Vladimír Brabec as Sendivoj (voice)

Jan Přeučil as Zimozel (voice)

Jiří Lábus as kouzelník Kelley (voice)

Miroslav Táborský as komoří (voice)

Petr Rychlý as Tycho de Brahe / náčelník (voice)

Anna Brousková as Reportérka Káťa Hvozdíková (voice)

Vladimír Javorský as Primář (voice)

Lucie Bílá as sestřička (voice)

Gabriela Osvaldová as Dr. Malebná (voice)

Zlata Adamovská as magistra Stříbrná (voice)

Martin Písařík as Tonda (voice)

Tomáš Juřička as Franta (voice)

Kryštof Hádek as Lojzík (voice)

Martha Issová as pošťačka (voice)

Ota Jirák as lev Aurix (voice)