LMPD said a passerby called police to report seeing what they thought was a human body floating in the water.
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The crash involved a semi and another vehicle, according to images from TRIMARC cameras.
One lane reopened just before 6 p.m., and all lanes had reopened by 8:15 p.m. Monday.
Interstate 264 will be closed in both directions from the I-264/I-64 interchange at mile point 18.7 to the I-264/I-71 interchange at mile point 22.7 from 7 a.m. to 5 p.m.
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The semi hit the wall on the interstate, causing the trailer to come unattached from the semi.
Right now on I-64 between the Gene Snyder and Simpsonville, there is a nine-mile stretch without an exit.
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According to LMPD spokesperson Dwight Mitchell, officers responded to a call of a vehicle collision around 1:30 a.m. on Interstate 64 East at mile marker 21.
Louisville police said the crash happened around 7:30 p.m. Tuesday.
I-64 westbound is currently scheduled to be closed until 4 p.m. at the I-264 interchange near St. Matthews to Grinstead Drive near Cherokee Park.
The busy stretch of interstate is one of the most heavily traveled in Kentucky.