After a reflection on the last year, accompanied by choirs singing the song, Tennant - one of the presenters on the Red Nose Day programme - began to well up and his voice was cracking as he urged viewers to donate.
“I’m crying too David Tennant,” tweeted one fan.
“David Tennant prob the most loved person in the UK,” said another.
A third added: “I’m sobbing david tennant is the most beautiful human being in this world.”
Earlier in the show, Staged stars Tennant and Michael Sheen reunited for a Zoom comedy sketch.
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Daniel Craig and Catherine Tate will appear later in the show for a James Bond sketch.
Find out everything you need to know about Comic Relief 2021 here.
Last year Tennant compared Doctor Who fame to “losing a layer of skin”. He starred as the Time Lord from 2005 to 2010, alongside Billie Piper as his companion Rose Tyler.
In an episode of David Tennant Does a Podcast With, the actor told Piper: “Entering into that madness... and particularly that loss of anonymity, which I had not really experienced before... because I had you to go on that journey with, I think I survived it much better than I might have done.”
He added: “You knew what it was to lose that layer of skin. It was very helpful to be on that journey with someone who understood.”
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