Julen Lopetegui has Andres Iniesta back in the Spain squad for the upcoming World Cup qualifiers against Italy and Albania, but there is no space for Juan Mata or Cesc Fabregas.
Barcelona captain Iniesta missed last month's international wins against Belgium and Liechtenstein with a knee injury but has since returned to fitness.
Spain take on Italy, the team that knocked them out of Euro 2016 in the round of 16, on Oct. 6 in Turin before facing a trip to Albania three days later.
"The return of Andres is great news for us," Lopetegui said at a news conference. "He was injured. We are happy to have him back. He has been playing regularly. We will need our best players to face Italy."
Lopetegui has included Napoli winger Jose Callejon as one of five changes.
Callejon, who last featured for Spain in November 2014, has been rewarded for an impressive start to the campaign with Napoli, having scored five goals in six Serie A appearances.
The return of Callejon sees forward Paco Alcacer, who has so far not made much of an impact since joining Barca this summer from Valencia, lose his place.
"Callejon was close to being included last month," Lopetegui said. "We think he is going through a good spell and we have decided to bring him."
Sevilla goalkeeper Sergio Rico and Real Madrid pair Isco and Nacho also return, despite both having limited playing time this season.
Concluye la conferencia de prensa de @julenlopetegui. Turno para Celades y la Sub-21 ¡Te recordamos la lista! pic.twitter.com/q8xoE0L3vE
- Selección Española (@SeFutbol) September 30, 2016
Asked about Isco, Lopetegui said: "It's not the same being a sub at Real Madrid than in another club. I can't deny that I like Isco. He has played 58 minutes in the league and that's very little. But he is a player that I like and that I trust. We have decided to bring him because he can help us in these games."
Madrid midfielder Marco Asensio has been named in Spain's under-21 squad, while Manchester United midfielder Mata, Adrian of West Ham and Chelsea's Cesar Azpilicueta are among those dropped.
"There are many players that we like but not all of them can come," Lopetegui said. "Adrian is not coming because Sergio [Rico] is playing at a great level."
On overlooking Fabregas, Lopetegui added: "Cesc is a great player, but not everyone can come."
Athletic Bilbao defender Aymeric Laporte was this week called up to the France squad amid talk that he could also be selected by Spain.
But Lopetegui said: "Laporte was never an option because he wasn't a player eligible for selection."
Spain demolished Lichtenstein 8-0 on Sept. 5 and are level on three points in their World Cup qualifying group with Italy, who won their opener 3-1 at Israel.
"We face two difficult away games," Lopetegui said. "We need to go into each game with the maximum ambition. They are two important games but they are not crucial. We know the challenges we will face but are enthusiastic."
Lopetegui succeeded Vicente del Bosque, who stepped down after eight years in charge following Spain's elimination from Euro 2016.
The full Spain squad:
Goalkeepers: David de Gea (Manchester United), Pepe Reina (Napoli), Sergio Rico (Sevilla)
Defenders: Nacho Fernandez (Real Madrid), Jordi Alba (Barcelona), Dani Carvajal (Real Madrid), Sergio Ramos (Real Madrid), Gerard Pique (Barcelona), Javi Martinez (Bayern Munich)
Midfielders: Koke (Atletico Madrid), Sergi Roberto (Barcelona), Saul Niguez (Atletico Madrid), Sergio Busquets (Barcelona), David Silva (Manchester City), Lucas Vazquez (Real Madrid), Thiago Alcantara (Bayern Munich), Andres Iniesta (Barcelona), Isco (Real Madrid)
Forwards: Jose Callejon (Napoli), Vitolo (Sevilla), Alvaro Morata (Real Madrid), Diego Costa (Chelsea), Nolito (Manchester City)