They claim the bike route, part of a £4.4million road improvement scheme, is hardly used and has literally driven away customers who no longer have spaces to park.
The traders' crisis follows 18 months of traffic chaos while the lane was being built.
Stephen Thomas, 60, who owns Smith's Bakery and Confectioners said he was forced to close one of his two shops in Castleton, Greater Manchester.
He said: "I had to let my staff go and I had to close the shop because it just wasn't paying anymore.
"Tradesmen used to jump out of their vans and people with mobility issues could park outside and come in. Unfortunately all of that has been taken away from us.
"My family has been trading in this village since 1928 and I'm the third generation to own this business.
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