The protesters handed out flyers while intermittently stopping traffic Thursday during the peaceful event, the Canadian Broadcasting Corp. reported.
The "Idle No More" protests center on perceptions that the Canadian government is moving to weaken environmental protections, particularly for waterways, to facilitate the construction of oil pipelines that would cross tribal lands.
Garden River First Nation member Darrell Boissineau said protesters worked with police to assure emergency vehicles could get through the blockade and for other vehicles to advance after about a 15-minute delay.
"The flow of traffic may be held up for 15 minutes and then it will be released but people know well in advance when the traffic is stopped and what they will be going through," Boissineau told the CBC.
"It may seem like a long 15 minutes, but what I've seen this morning, it seems to be well-organized."
Similar protests were carried out on other highways in northeastern Canada during the holiday period and Attawapiskat Chief Theresa Spence reached the 17h day of her hunger strike in Ottawa in her effort to get the government to negotiate with the tribes over the environmental issues.
"I'm really proud of youth who are standing up to the government and to your community member and saying that you want a healthy life and that we deserve a healthy life and a safe environment in our lives," she said.