The objective was to determine whether increased dietary protein would enhance mammary development in prepubertal heifers fed for rapid body growth (1.2 kg/d). Fifty-four Holstein heifers (weighing approximately 134 kg) were assigned to one of three treatments. Heifers were fed a total mixed ration with metabolizable energy at 2.85 Mcal/kg and metabolizable protein at low, standard, or high concentrations (37, 41, or 44 g/Mcal of metabolizable energy, respectively) from 3.5 mo of age until slaughter at approximately 46 d after puberty. Heifers fed low, standard, and high protein gained 1130, 1170, and 1180 g/d, respectively. Dietary protein did not affect age or weight of heifers at puberty or slaughter, withers height gain, or carcass composition. Average mammary parenchymal DNA content for heifers on diets of low, standard, and high protein was 595, 619, and 670 mg/100 kg of body weight, respectively, and was not significantly different. However, for heifers that attained puberty early, those fed low protein had 33% less parenchymal DNA than those fed high protein, even though their body growth and carcass composition were not compromised. We conclude that dietary protein does not have a major effect on mammary development of rapidly grown prepubertal heifers, provided the diet contains adequate protein for normal body growth. But we suggest that feeding low-protein diets increases the risk of impaired mammary development when heifers are fed for rapid growth and attain puberty early and that the new National Research Council guidelines for protein relative to energy seem adequate for optimal mammary development.