The 224-209 vote against the $142.5 billion spending bill disrupted plans by Republican leaders to finish up work on 11 spending bills funding government operations.
"The health care delivery system in rural Kansas and Missouri is very fragile," said Rep. Jerry Moran, R-Kan., who voted against the measure. "There has to be somebody that steps forward and says there is a real problem here in nearly zeroing out a number of programs related to rural health care."
CBS News correspondent Bob Fuss called the bill's rejection an "embarrassing and unexpected loss for House Republican leaders."
That's at least in part because the last time the House Republican leadership lost a vote on the final version of a major spending bill was 1995, Fuss reports.
In the Senate, Republicans beat back Democratic attempts Thursday to use a $60 billion tax bill to pinch oil and energy companies that have been reporting record profits while consumers pay high gasoline prices.
Fuss adds that the Senate will probably pass the bill, but that the House has not passed its version of a tax cut bill, which will be quite different.
The bill, which would prevent a number of individual and business tax breaks from expiring, already levies more than $4 billion in taxes on major oil companies.
House Democrats were unanimous in opposing the one-year appropriation bill, which included the first cut in education funding in a decade and slashed spending for key educational programs such as Head Start and No Child Left Behind.
But it was the 22 GOP defections that sent it down to defeat.
Rep. Jo Ann Emerson, R-Mo., said she was "fed up" with the dramatic cuts that would impact her rural district in southeastern Missouri. Funding for rural health research, for example, would drop from $8.8 million annually to zero. The bill would also eliminate a half million dollars in funding for rural emergency medical services and cut funds to attract health care professionals to rural areas.
"I see a huge erosion in the support for rural health care," she said. "I cannot sell out my district."
Other Missouri and Kansas Republicans all voted in favor of the bill.
Some Republicans speculated they may have lost votes because this year's bill included no special projects or earmarks for lawmakers. "You take those out and you lose the incentive," said Rep. Tom Davis, R-Va., who voted for the bill.
Democrats, meanwhile, hailed the vote as a major defeat for Republicans.