US Defence Secretary Leon Panetta says a video which appears to show US Marines urinating on the corpses of Afghans is "utterly deplorable".
Those who had taken part in the incident would be held accountable "to the fullest extent", he said.
The video, which was posted online, purports to show four US Marines standing over the bodies of several Taliban fighters, at least one of whom is covered in blood.
The origin of the video is not known.
Mr Panetta said he had ordered the commander of US and Nato forces in Afghanistan to investigate the incident.
"I have seen the footage, and I find the behaviour depicted in it utterly deplorable," he said in a statement.
The Pentagon says it is checking the authenticity of the video, but there is nothing to indicate that the film is not genuine.
The BBC's Steve Kingstone, in Washington, says senior officials have wasted no time in going into damage-limitation mode.
Our correspondent says the alleged incident is deeply embarrassing for a US administration that had promised to project a more sensitive image of American military power.
It also comes at a time when tentative moves are being made towards peace, ahead of the planned withdrawal of international combat forces by the end of 2014.
The Taliban said last week that they were working to set up a political office, possibly in Qatar, that would help to facilitate negotiations with the Afghan government and Nato countries.
The video has not yet been circulated widely in Afghanistan, but there are fears that it could provoke further violence against international forces.
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