The Premier League has written to Wolves asking for an explanation after manager Mick McCarthy made 10 changes to his side to face Manchester United.
McCarthy decided to change all of his outfield players from Saturday's 1-0 win over Tottenham Hotspur for the trip to Old Trafford on Tuesday evening.
Wolves lost 3-0 at the hands of the champions and sportsbook second favourites for the Premier League title.
Under league rules, every club has an obligation to field its strongest possible team in order to maintain the integrity of the competition.
"The Premier League Board have written to Wolverhampton Wanderers to request their observations in relation to the team fielded in their League fixture against Manchester United," confirmed a statement.
"Once the observations are received the Board will decide whether any further action is warranted."
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Renault has ended the speculation surrounding its Formula One team after confirming they will definitely be on the grid for the 2010 season.
The French team has struck a deal with Luxembourg businessman Gerard Lopez and his Genii Capital investment company which has secured their future in the sport.
Renault, who had considered ending their lengthy association with Formula One, will retain a 25 per cent stake in the team, which will still be known as Renault F1.
Meanwhile, Mercedes chief executive Nick Fry has dropped a strong hint that Michael Schumacher will come out of retirement to race for them next season.
The seven-times world champion is seen as a perfect fit for the new guise of the Brawn GP team and is widely expected to end his role as a consultant with Ferrari to make the switch before the start of the 2010 Formula One season.
Harlequins have confirmed the appointment of Conor O'Shea as their new director of rugby.
The former London Irish and Ireland player is the replacement for Dean Richards, who was banned for his part in the 'Bloodgate' saga.
Meanwhile, Delon Armitage has declared himself fit to return to action after a shoulder injury suffered in September had threatened to end his career.
The 27-year-old London Irish player has been a key figure for both club and country over the past 18 months, and his return will be heralded in both camps.
The full-back dislocated his shoulder against Gloucester in the Guinness Premiership back in September and was forced to sit out England's much maligned autumn campaign.