Some of Pete's lines from this short are also heard in "Get a Horse!".
There is a video of both Walt Disney and Billy Bletcher in a recording session for this short.
In this short, Mickey's shoes are brown rather then the trademark yellow.
The train station that Mickey and Pluto depart from is located in Burbank, California which also happens to be the home of Walt Disney Productions headquarters.
"Mr. Mouse Takes a Trip" from 1940 is unique among Disney shorts in that film footage exists of the voice-over session, which included Walt Disney and Billy Bletcher. According to film historian Leonard Maltin, the footage was not known to exist and only discovered (as of 2004) "not too many years ago". The black-and-white film, which is about ten minutes in length, is the only known footage of Disney performing as Mickey Mouse on Camera. It was included as an extra on the 1997 VHS and 2000 DVD of "Bongo e i tre avventurieri (1947)" from 1947, and on the 2004 DVD release "Walt Disney Treasures: Mickey Mouse in Living Color, Volume Two" as an easter egg.