Dozens of high school and college students rallied at the Torch of Friendship in Miami before boarding a bus to Tallahassee to protest against what many opponents have termed the "Don't Say Gay" bill.
The Florida Senate is scheduled to vote on the bill Monday.
"There are so many LGBT youths that are going to have their lives put at risk," one student said.
Miami-Dade County Mayor Daniella Levine Cava and other elected officials sent them on their way.
"It's shameful what is going on in Tallahassee. So many ways that they are trying to take away our rights, our human rights, and these young people are getting on a bus and riding all night to tell them, 'Enough is enough,'" Cava said.
The bill, formally called the Parental Rights in Education bill, prohibits classroom instruction on sexual orientation or gender identity in kindergarten through 3rd grade, though some fear the vagueness could make it broader.
One clause in the bill reads, "Classroom instruction by school personnel or third parties on sexual orientation or gender identity may not occur in kindergarten through grade 3, or in a manner that is not age-appropriate…"
Many Republican lawmakers insist that curriculum in early grades not include content about sexual orientation and gender identity.
Those matters, they say, should be discussed at home and not in school.
"I, as a queer person, feel silenced by the use of the 'Don't Say Gay' bill," one student said. "I hope to let my representatives know that this is not okay."
Supporters of the bill say that the headlines are not what they seem.
"If you read the bill, it is protecting parents rights and we're talking about kids in kindergarten through third grade. They shouldn't be talking about those topics. That's all it is," state Sen. Manny Diaz said.