Freddy Guevara urged people to protest tomorrow -- May 1 or May Day, a traditional workers' holiday -- with marches to the Supreme Court and the National Electoral Council's offices.
"We want to summon all Venezuelans, across the country's 24 states, to hit the dictatorship with a one-two punch," Guevara said, evoking simultaneous peaceful marches in the two locations.
A month of demonstrations in April in Venezuela has left 28 people dead in clashes between riot police and anti- government protesters, according to prosecutors.
The center-right-led opposition is demanding elections to remove Maduro, an elected socialist. It blames him for an economic crisis that has caused shortages of food, medicine and other basic items.
"On May 1, after a month of resistance, we need to show that the people refuse to give up," Guevara said.
Rights group Amnesty International urged the government on Wednesday to stop the "persecution" and "arbitrary detention" of protesters.
Earlier in the day, Pope Francis said the Vatican was ready to intervene as a mediator to try and end Venezuela's deadly political crisis under "clear conditions".
"There was an intervention by the Holy See following strong pressure" by four former leaders of Spain, the Dominican Republic, Panama and Colombia, he said.
"This did not work because the proposals were not accepted. They were diluted. It was a 'yes, yes but no, no'", he said.
The pontiff said the four mediators were trying to relaunch the process, adding "they are looking for a venue" but the opposition was against the process.
"We should do all that we can for Venezuela with necessary guarantees," he said.
Just Wednesday Venezuela said it was quitting the Organization of American States in anger at pressure from the bloc over the government's handling of a its political crisis.
Echoed by the United States and European Union, the OAS has led an international chorus of concern over the economic and political chaos in the major oil-exporting country.
Maduro says the shortages and the protests are part of a US-backed plot to topple him.
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