The largest five-digit number that can be formed using 0, 1, 2, 3, 4 exactly once is:
\[
\text{Maximum number} = 43210.
\]
The smallest five-digit number that can be formed using 0, 1, 2, 3, 4 exactly once is:
\[
\text{Minimum number} = 10324.
\]
The difference between the maximum and minimum numbers is:
\[
43210 - 10324 = 32976.
\]