1. 1. We studied the lipid composition and the fluidity of small intestine brush border membrane (BBM) of rats of different age: ‘very young’ (5–7 weeks old), ‘young’ (9 weeks old), ‘adult’ (30 weeks old) and ‘old’ (85 weeks old). 2. 2. Fluorescence anisotropy, as assessed by 1,6-diphenyl-1,3,5-hexatriene probe (DPH), was increased from very young to adult rats. 3. 3. In agreement with these results the lipid composition in adult animals showed a lower lipid/protein ratio (derived mainly from a lower content of total polar lipids) and an increase of cholesterol esters and sphingomyelin (SM) saturation index. 4. 4. A marked decrease of the order parameter was observed in the ‘old’ group, accompanied by a decreased cholesterol/phospholipid ratio. 5. 5. The percentage distribution of membrane phospholipids significantly changed during development, but the modifications were not correlated with the anisotropy of DPH.