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Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) have been implicated in normal brain development, adult stroke, and, more recently, perinatal brain injury. Here, our objective was to obtain comprehensive and comparative data on the ontogeny of MMP-2, MMP-9, TIMP-1, and TIMP-2 in the neocortex of male and female mice belonging to various strains, from embryonic life to adulthood. We used gelatin zymography, ELISA, and real-time PCR analyses. MMP-2, MMP-9, and TIMP-1 activity and/or expression peaked during embryonic life and the early neonatal period, whereas TIMP-2 peaked during the first two postnatal weeks. Comparable results were obtained in all the mouse strains except BALB/c, where MMP-2 levels were considerably lower at all ages compared with the other strains. No gender effect was observed on any of the study parameters. This comprehensive study will serve as a basis for future investigations into the role for MMPs and TIMPs in normal brain development and prenatal brain injury.