Yes, and actually there are a lot of differences.
One of them is that in Spain people say "Vosotros" when they talk about "all of you", and in Mexico (as in all the Latinamerican countries) it's used the word "Ustedes" that has a different way to conjugate the verbs.
Vosotros sois = ustedes son
There also are differences between these (Spanishes?) and the Uruguayan/Argentinian Spanish.
There, people don't say "Tú", or "Usted".. they say "Vos", that also has a different way to conjugate the verbs
Tú eres = usted es = vos sos
Tú tienes = usted tiene = vos tenés
Obviously there also is a very different pronunciation in each country, in Uruguay our lenguage is very.. mm.. "regular" (it's almost always the same) but in Mexico they talk like if they were "singing" (for us), and in Spain they talk like if they were punishing you or something :P
And of course different vocabulary too:
In Spain the verb "coger" means "to take something", but in Uruguay or Argentina, it means "to f*ck". In Spain "¡joder!" means "f*ck!" but in Uruguay/Argentina it means "to annoy someone" or "kidding".
Sorry if my English has some mistakes but I'm from Uruguay :)