Matrix metalloproteinases (MMPs) have been implicated in tissue remodelling and in regulation of cell-matrix interactions during organ development. The activity of MMPs is regulated by members of the TIMP (tissue inhibitors of metalloproteinase) family. We analyzed by in situ hybridization the expression of gelatinase A (MMP-2) and gelatinase B (MMP-9) as well as Timp-1, -2 and -3 during different stages of mouse tooth development. Gene expression was generally found in mesenchymal tissues except for Timp-3, which also was found in dental epithelial cells. During early tooth development, gelatinase A and Timp-2 were widely expressed in the branchial arch, while gelatinase B and Timp-1 and Timp-3 expression showed clear association with epithelial morphogenesis and was restricted to the mesenchyme at the tip of the growing tooth bud. Gelatinase A and Timp-1 showed transient expression in secretory odontoblasts at the time of basement membrane degradation, while Timp-2 expression continued throughout the dental papilla. At the time of tooth eruption, Timp-3 was expressed in most dental epithelial cells except secretory ameloblasts, and gelatinase B was intensely expressed in osteoclasts in the jaw bone. The exact co-localization of gelatinase A and Timp-1 in secretory odontoblasts, and the correlation between gelatinase B and Timp-3 during bone resorption may indicate interaction of the proteins during degradation of the basement membrane and in the control of ECM turnover in connection with tooth eruption.