We offered 43 Holstein-Freisian dairy heifers that calved for the first time at either 2 or 3 yr of age ad libitum access to ryegrass silage and a standard concentrate allowance of either 2 or 7 kg/d from the middle to the end of their first lactation. All animals were given the same relatively poor quality dry period diet of a mixture of ryegrass silage and barley straw (63:37 dry matter basis) from 6 wk before their predicted second-calving date. Following their second calving, all animals received access to the same ration of ad libitum grass silage and concentrates at a rate of 8 kg/d to 120 d of lactation and 5 kg/d thereafter until the end of the experimental recording at about 150 d of lactation. Nitrogen balance was significantly higher at the end of the first lactation for animals that were given the higher concentrate allowance and tended to be higher for older animals. There was no effect of age or residual effect of concentrate allowance on N balance during the dry period or during the second lactation. Labile body protein reserves, as estimated by the depth of the muscle Longissimus dorsi (which was significantly correlated with body condition score), were similar for all animals during the dry period, but younger animals previously offered the lower concentrate allowance did not lose L. dorsi depth early in the second lactation as did other animals. Arterial plasma concentrations of amino acids Phe, Trp, and Leu were significantly higher in younger animals at wk 8 of the second lactation, and Gly was significantly lower, although mammary blood flow, arteriovenous differences, and rates of uptake of the AA measured were unaffected by treatment. It is concluded that differences in second-lactation milk yield were not mediated through the availability of labile body protein or the supply of nutrients to the mammary gland.