Joe Anderson said Scots should vote No and work together with cities like Liverpool instead of voting for independence.
Mr Anderson, a Labour supporter, said if cities across the UK had more power over investment and development, they could create benefits for each other, while remaining within the UK.
He said: "I question what difference it will make to cities like Glasgow if they become independent.
"It will disconnect cities like Glasgow and Liverpool when we should be better connected. I want better transport links, links though our universities and our businesses.
"To split the UK wouldn't benefit Scotland or the rest of the UK. We should fight for more devolution."
Mr Anderson also questioned First Minister's Alex Salmond's understanding of Glasgow.
Speaking outside the hall where Mr Salmond was addressing business leaders, he said the links between Liverpool and Glasgow were as strong as any that existed within Scotland and said a walk around either city was enough to confirm that view
He said: "I have more in common with Glaswegians than Alex Salmond. They are people who are proud of their city and Liverpool people are as well.
"I don't think Alex Salmond knows what the needs of Glasgow are any more than David Cameron does.
"It is the people in Glasgow who know best how to shape the city, more than the Scottish Government."