Some 470 parliamentary staff have signed a petition criticising the decision to convert a Commons bar into a nursery.
Presenting the petition to the Commons late last night, Tory MP Christopher Chope said the collection of so many signatures over just two days demonstrated the "depth of concern about the proposals".
Parliamentary authorities are planning to turn Bellamy's bar on the Commons estate into a crèche, designed to provide child care for up to 40 children.
The move, supported by John Bercow, aims to make Parliament more family friendly and encourage a wider variety of people to become MPs.
But some MPs and staff are unhappy at seeing one of their watering-holes closed when they believe other suitable empty accommodation is available.
Earlier in the day Chope had raised a point of order with the Speaker in the Commons, claiming that staff had been unfairly urged not to sign the petition.
"It has been drawn to my attention that the departmental communications officer of the Department of Facilities on Friday sent an email to staff saying that they should express their views through their line manager and by implication not sign the petition," he said.
"Please will you confirm that the democratic right to petition Parliament extends to all staff working in Parliament and they are free to sign the petition without fear of victimisation."
Bercow said he would "make inquiries" into the matter.
"I stand by what I said about the right to petition - I will look further into this matter and the very particular charge of criticism that you make," he added.
Expressing his support in December for the creation of Parliament's first nursery, Bercow said it was time the House of Commons "caught up" with the rest of the world.
"Our Parliament has sadly been behind the times in providing practical support to parents who work here. If Parliament is to be truly representative of the community it serves then it must do more to encourage parents to stand as MPs.
"Many other Parliaments around the world provide nursery facilities for members and staff. It is time one of the oldest Parliaments in the world caught up with the rest. I am delighted that the Commission has taken this historic decision."
But the petition argues that the estimated £400,000 cost of conversion is "unacceptable in the present economic climate."
Signatories are also unhappy that authorities appeared to backtrack on a commitment to consult the Commons administration committee, of which Chope is a member, on the proposals.
Chope's petition calls for the nursery to be located in the Derby Gate building, located a few minutes walk from the House of Commons, just off Parliament Street.
The space is currently earmarked for conversation in to accommodation for use by MPs and their staff as part of a series of parliamentary office moves.
The Derby Gate premesis is currently occupied by Department of Information Services staff, who are moving into 7 Millbank, offices that are soon to be vacated by staff who deal with MPs allowances and are being transferred to the Independent Parliamentary Standards Authority.
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