Current data indicates that feeding dairy calves more nutrients in early life allows them to produce more milk in the future. Mechanisms responsible are poorly understood. Thirty-six Holstein heifer calves were fed either a restricted (R; 20.2% crude protein [CP], 19.8% fat, dry matter (DM) basis, fed at 0.44 kg/hd/d, DM basis) or an enhanced (EH; 28.9% CP, 26.2% fat, DM basis, fed at 1.08 kg/hd/day, DM basis) milk replacer (MR) and given either a placebo or estradiol (E2) implant to assess differential responses to E2. Our underlying hypothesis was that calves fed more nutrients are better able to respond to mammogenic stimuli and will have a more developed mammary gland as a result of imposed treatments. Enhanced-fed calves grew at a faster rate, were heavier at weaning, and had more functional mammary tissue (i. e., parenchyma; PAR) mass in the mammary gland at weaning (7.3-fold). Additionally, biochemical composition of the PAR was not impacted by the dietary treatments imposed. Furthermore, EH-fed calves had an increase in the number of actively dividing cells throughout the mammary PAR as well as increased intensity of estrogen receptor expression in the population of cells expressing the estrogen receptor. Enhanced-fed calves had an up-regulation of genes and pathways in the PAR related to metabolism, cellular signaling, and cellular growth. When given E2, EH-fed calves experienced the greatest overall mammary gland development and had the greatest PAR mass without compromised composition. When comparing EH- and R-fed calves given E2, differential expression of genes and pathways related to cell growth, cell signaling, and metabolism was observed. In summary, data indicates that enhanced feeding of calves in early life allows increased responsiveness to mammogenic stimuli and a corresponding increase in mammary development. We suggest that this may at least partly explain the improved future milk production in calves fed in this manner.