In 1972, the show was re-conceptualized as Josie and the Pussycats in Out Space, 16 episodes of which aired on CBS during the 1972-73 season and were rerun the following season until January 1974. Reruns of the original series alternated between between CBS, ABC, and NBC from 1974 through 1976. This brought its national Saturday morning TV run on three networks to six years.

Josie and the Pussycats featured a teenage all-girl group pop band the foured the world with their entourage, getting mixed up in strange adventures, spy capers, and mysteries. The group consisted of level-headed lead singer, songwriter and guitarist Josie, intelligent tambourine Valerie, and air-headed blonde drummer Melody. Other characters included their cowardly manager Alexander Cabot III, his conniving sister Alexandra, her cat Sebastian, and muscular roadie Alan.

The Show, more similar to Scooby-Doo, Where Are You! than the original Josie comic book, is remembered for its music, the girls' leopard print leotards (replete with "long tails and ears for hats", as the theme song states), and for featuring Valerie as the first regularly appearing female black character in a Saturday morning cartoon show. Each episode featured a Josie and the Pussycats song played over a chase scene, which, similarly to The Monkees, featured the group running after and away from a selection of haplessly villainous characters.

Josie and the Pussycats debuted on the CBS Saturday morning lineup on September 12, 1970, with the episode "The Nemo's a No-No Affair". The animated versino of Josie was an amaigam of plot devices, villain types, settings, moods, and tones from other Hanna-Barbera shows such as Scooby-Doo, Where Are You!, Jonny Quest, Space Ghost, and Shazzan.

Like Scooby-Doo, Where Are You!, Josie and the Pussycats was originally broadcast with a laugh track. Later home video and DVD releases omit the laugh track. Cartoon Network and Boomerang, however, have aired the show in its original broadcast format with the laugh track intact.

Each episode found the Pussycats and crew en route to perform a gig or record a song in some exotic location where, somehow, often due to something Alexandra did, they became mixed up in an adventure. The antagonist was always a diabotical mad scientist, spy, or criminal who wanted to take over the world using some high-tech device. The Pussycats usually found themselves in possession of the plans fo an invention, an item of interest to the villains, a secret spy message, etc., and the villains chased them to retrieve it. Eventually, the Pussycats would ruin the villain's plans, resulting in a final chase sequence set to a Pussycats song. With the villain captured, the Pussycats would return to their gig or recording session, and the final gag was always one of Alexander's failed attempts to interfere with the Pussycats' performance or steal Alan away form Josie.