Adapted from an Archie spin-off comic, the animated version differs from the actual comic most significantly in the characters of Alexandra, Sebastian, and Alexander. In the cartoon, Alexandra and Sebastian are simply conniving and sneaky; in the comic book, Sebastian is the reincarnation of a witchcraft-practicing ancestor of the Cabots, and Alexandra can cast powerful magic spells while she is holding him. In the original Archie comic, Alex is a snide braggart who lords his wealth over everyone and sees Alan M. as his rival for Josie's attention. In the animated series, Alex is a meek coward who brags much more about the band's talent than his wealth, and expresses no romantic interest in Josie.
Cheryl Ladd (credited as Cherie Moor) provided the singing voice for Melody Valentine. Patrice Holloway, sister of Motown singer Brenda Holloway, provided the singing voice of Valerie and sings lead on the theme song.
Hanna-Barbera wanted to change Valerie to a Caucasian even though she was already established as African-American in the Archie comics, so they wanted to fire Patrice Holloway who had been cast as Valerie's singing voice. But Danny Janssen, who was producing the real-life group's Capitol album, refused to fire Holloway and Hanna-Barbera relented.
As with Scooby-Doo, dove sei tu? (1969), Josie and the Pussycats originally aired with a laugh track. Remastered prints omit the laugh track. The second season of Scooby-Doo, produced concurrently with Josie and using much of the same creative team, features musical interlude "chase songs" just as Josie does.