Daniele De Rossi, Giorgio Chiellini and Andrea Barzagli all said they would retire from international duty after Italy failed to qualify for the World Cup on Monday.
In stepping away after the goalless draw with Sweden, they will join goalkeeper Gianluigi Buffon, who had previously said he would be done following the World Cup.
De Rossi, 34, scored 21 international goals over his career, and his 117th cap on Monday moved him past Andrea Pirlo into fourth place on Italy's all-time appearance list.
"It's a dark moment for our football, very dark for those of us who were part of this two-year era," De Rossi told RAI. "There will be the time to evaluate everything, but I think the only thing I can say now -- which is banal -- is to build on the spirit the lads showed today and the desire to take part in this adventure.
"We will begin again, just as we did after other desperately disappointing moments. I don't believe we deserved to go out considering the 180 minutes, Sweden deserve credit for their performance, but it was fairly even.
"Now the next generation is ready to take flight and we must begin again from them. It was almost an absurd moment to associate with a football match. There was a funereal atmosphere in the locker room, yet nobody died.
"I've been wandering around Coverciano [the training ground] and all over the world with this shirt for over a decade, so to take it off for the last time is a strange feeling. The material is there and we can build for the future."
De Rossi was spotted arguing with his coach over a substitution, urging Gian Piero Ventura to put on forward Lorenzo Insigne as the Italians needed a late goal.
"We have this tendency to warm up three at a time, then after five minutes we change and another three go," he explained. "I just said we were near the end and had to win, so send the strikers to warm up! I pointed to Insigne too.
"It wasn't up to me whether it was a tactical issue and the prof is a lovely guy, so I'm sorry if I offended anyone. At the time I just thought perhaps it was better that Insigne come on instead."
Chiellini, 33, made 96 appearances for Italy, and the defender found the net eight times, and he encouraged fans to embrace the next generation of players.
"We have many young lads who over the next few years will need this same love and support," he said. "It's a long road ahead and after a fall like this there is much work to be done.
"Over the last three years or so we have asked for this love to never leave the Nazionale. Italy will begin again from Marco Verratti and many other lads born in the 1990s who will be mature for the next Euros, which fortunately we'll have some games at home.
"I am certain that if what we saw tonight can be built on, it will be seen as a Zero Hour. Italy hadn't missed the World Cup in 60 years, so clearly it is a sea change. Football belongs to everyone and we all want to create a new era."
Barzagli, 36, said it was an "end of an era" after he appeared in 73 international games.
"In football terms, it's the biggest disappointment of my life," he said. "The Italy side gives you unique sensations. It gives you goosebumps, creates friendships between all the Italy fans, they are finally united. It's a real shame to end like this.
"I don't know what we missed, all I know is we're out of the World Cup, it's a unique disappointment and leaving this group of lads is painful. Everyone has their own feelings, but it's really hard to accept right now.
"At least at Euro 2016 we were crying because we came so close, but not even qualifying is a heavy blow for my whole career. With Gigi and Daniele we had the adventure in the 2006 World Cup, but also had many adventures with Giorgio.
"It's immensely disappointing. I hope the younger players can take the situation in hand and do better than us. The era of four or five veterans comes to a close, the one of the hungry young players coming through begins and that's how it should be."