"The Origin of the World", a nude painted from 1866, was protected by a "glass pane" and the police were on site to assess the damage, the Centre Pompidou in the northeastern city of Metz told AFP.
The work had been on loan to the Centre Pompidou-Metz from the Musee d'Orsay in Paris as part of an exhibition centred on French psychoanalyst Jacques Lacan, who once owned the painting.
Metz prosecutor Yves Badorc said two women born in 1986 and 1993 had been arrested after five works in total, including Courbet's nude, had been sprayed with the words "MeToo".
A third person -- who has not been detained -- is believed to have stolen another artwork, he said.
The stolen piece -- red embroidery on white material by French artist Annette Messager -- is called "I Think Therefore I Suck".
- 'Women are the origin of the world' -
French-Luxembourgish performance artist Deborah de Robertis told AFP she had organised the spray painting in red, carried out by two other people, as part of a performance titled: "You Don't Separate the Woman from the Artist".
In a video sent to AFP by de Robertis, one woman tags Courbet's famous painting with red paint and then another one.
They then chant "MeToo" before being dragged away by security guards.
De Robertis explained that she wanted to "challenge the history of art", in particular by tagging "MeToo" on the famous painting "because women are the origin of the world".
In an open letter, de Robertis denounced the behaviour of six men in the art world, describing them as "predators" and "censors".
She said the actions were a feminist performance, carried out because "the very closed world of contemporary art has remained largely silent until now".
The artist said they had also targeted another work by Austrian artist Valie Export.
De Robertis already had work on display at the venue -- a photograph of a 2014 performance at the Musee d'Orsay in which she posed showing her vagina underneath Courbet's painting.
- 'Attack on culture' -
Culture Minister Rachida Dati said five works had been targeted.
"To 'activists' who think that art is not powerful enough to carry a message alone... An artwork is not a poster to colour in with the day's message," she wrote on X, formerly Twitter.
Metz mayor Francois Grosdidier condemned what he described as "a new attack on culture, this time by fanatic feminists".
Courbet's nude was first owned by a Turkish-Egyptian diplomat called Khalil Bey, a flamboyant figure in 1860s Paris who put together an art collection celebrating the female body before he was ruined by his gambling debts, according to the Musee d'Orsay.
It belonged to Lacan before it entered the museum's collection in 1995.
A French court in 2020 sentenced de Robertis to pay a 2,000-euro ($2,150) fine for appearing naked in 2018 in front of a cave in the town of Lourdes in southwest France, a Catholic pilgrimage site for those who believe the Virgin Mary appeared there.
A case against her was dropped in 2017 after she showed her vagina in front of Leonardo da Vinci's Mona Lisa at the Louvre museum in the French capital.
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