This work was designed to study the effect of a 3-day mild hyperglycemia (5.3 vs. 3.3 mM) on the regulation of glucose metabolism in lactating goats. Glucose was intravenously infused at variable rates simultaneously with a constant potassium-amino acid infusion. Diet plus substrate infusion maintained net energy but not protein supply. Milk yield did not change. Skeletal muscle glucose transporter (GLUT-4) was analyzed before and after hyperglycemia. In addition, the acute effect of medium and high insulin doses on glucose turnover was measured in vivo during euglycemic and hyperglycemic hyperinsulinemic clamps under potassium and amino acid replacement. Hyperglycemia reduced the endogenous glucose appearance but increased glucose disposal. It decreased the total membrane-associated GLUT-4 protein in skeletal muscle. In contrast, it improved the acute insulin-stimulated glucose disposal. Both the level and duration (3 days) of hyperglycemia contributed to this improvement. We conclude that short-term mild hyperglycemia has similar effects in lactating goats as those already observed in nonlactating rodents or humans.