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Spain under-21 manager Julien Lopetegui on Thursday called up full internationals Isco and Thiago Alcantara as well as Manchester United's David de Gea for the European Championships next month in Israel.
Thiago and De Gea were also part of the squad that won the title in Denmark two years ago and their involvement means they and Isco will not be included in Vicente del Bosque's senior squad for the Confederations Cup which also takes place in June.
"La Furia Roja" have been unbeaten in competitive matches at under-21 level since losing to the Netherlands in 2009, as they also cruised through their qualifying group for the tournament for the loss of just two points from a possible 24.
The squad will meet up at the Spanish Football Federation's headquarters in Madrid on Monday for a week's training but the players will be released to play for the clubs for the final weekend of the Spanish league season on June 1.
They will then travel to Israel and begin the defence of their title against Russia on June 6 before going on to also face Germany and Holland in Group B.
Spain under-21 European Championship squad:
Goalkeepers: David De Gea (Manchester United), Joel Robles (Wigan), Diego Marino (Villarreal)
Defenders: Nacho Fernandez (Real Madrid), Martin Montoya, Marc Bartra, Marc Muniesa (Barcelona), Ínigo Martínez (Real Sociedad), Alberto Moreno (Sevilla), Dani Carvajal (Bayer Leverkusen), Alvaro Gonzalez (Real Zaragoza)
Midfielders: Koke (Atletico Madrid), Asier Illarramendi (Real Sociedad), Thiago Alcantara (Barcelona), Isco, Ignacio Camacho (Malaga), Iker Muniain (Athletic Bilbao), Pablo Sarabia (Getafe), Sergio Canales (Valencia)
Strikers: Alvaro Morata (Real Madrid), Cristian Tello (Barcelona), Rodrigo (Benfica), Alvaro Vazquez (Getafe)