The National Court in Madrid summoned Josep Luis Trapero and three other suspects to a hearing on Friday October 6th, said the spokesman, who asked not to be named.
The accusations refer to unrest in Barcelona on September 20 and 21 after national security forces raided regional government offices in a crackdown against the independence drive.
That police action was met with furious protests and the Catalan regional police were accused of failing to intervene.
Tensions between Catalonia and the national government have plunged Spain into its worst political crisis in decades.
The spokesman said the four suspects were accused of “a crime of sedition… in relation to the gatherings and demonstrations carried out to forcibly prevent the authorities and their officers from carrying out their duties.”
The crime of sedition is punishable by up to 15 years in prison if committed by a member of the authorities, under Spain's penal code.
The judge leading the judicial investigation, Carmen Lamela, also summoned the leaders of two prominent Catalan pro-independence civil groups: Jordi Cuixart of Omnium Cultural and Jordi Sanchez of the Catalan National Assembly (ANC).
Those two groups were involved in the September demonstrations. Protesters damaged Civil Guard police vehicles and stopped officers from leaving the building they were searching.
The fourth suspect placed under investigation was another senior Catalan police official, Teresa Laplana. The region's leaders defied Madrid by holding a banned referendum on independence on Sunday.
National security forces beat unarmed voters as officers shut down some polling stations.