Age-dependent outcomes following traumatic brain injury motivate the study of brain injury biomechanics in experimental animal models at different stages of development. Finite element models of the rat brain are used to better understand the mechanical mechanisms behind these age-dependent outcomes; however, age- and region-specific rat brain tissue mechanical properties are required for biofidelity in modeling. Here, we have used the atomic force microscope (AFM) to measure region-dependent mechanical properties for subregions of the cortex and hippocampus in P10, P17, and adult rats. Apparent elastic modulus increased nonlinearly with indentation strain, and a nonlinear Ogden hyperelastic model was used to fit the force-deflection data. Subregional heterogeneous distributions of mechanical properties changed significantly with age. Apparent elastic modulus was also found to increase overall with age, increasing by between P10 and adult rats. Unconfined compression tests were performed on whole slices of the hippocampus and cortex of P10, P17, and adult rats to verify the mechanical properties measured with the AFM. Mean apparent elastic modulus at an indentation strain of 30% from AFM measurements for each region and age correlated well with the long-term elastic modulus measured from 30% unconfined compression tests (slope not significantly different from 1, ). Protein, lipid, and sulfated glycosaminoglycan content of the brain increased with age and were positively correlated with tissue stiffness, whereas water content decreased with age and was negatively correlated with tissue stiffness. These correlations can be used to hypothesize mechanistic models for describing the mechanical behavior of brain tissue as well as to predict relative differences between brain tissue mechanical properties of other species, at different ages, and for different regions based on differences in tissue composition.