The authors studied the spatial expression and regulation of messenger RNA for the alpha subunit of collagen type I in crushed rat sciatic nerve to provide a basis for future therapeutic manipulation. Sciatic nerves in 20 male or female adult Lewis rats were crushed for 60 seconds; the unharmed contralateral sciatic nerves served as controls. Twenty-one days after injury the experimental animals were killed and their tissue was harvested. The spatial expression of collagen type I was determined by using in situ hybridization techniques. Quantification of fibroblast number and total signal was performed through computerized morphometry. Collagen upregulation was evident in epineurial and perineurial layers, with the epineurium displaying higher activity. The cells responsible for procollagen type I production were fibroblasts. No activity was seen in the endoneurium. Morphometric findings indicated that collagen upregulation in the epineurium and perineurium occurred at both pretranscriptional and posttranslational levels when compared to controls; a paired t-test analysis confirmed statistical significance for all comparisons between injured and control tissues. Epineurial fibroblasts are responsible for the collagen production associated with crushed peripheral nerve injury in the rat. Regulation occurs pretranscriptionally as well as posttranslationally. It is interesting to speculate that the delivery of agents directed against collagen production (such as neutralizing antibodies to growth factors) into epineurial tissues proximate to the time and location of clinical nerve injury might mitigate later deleterious effects of excess collagen production in axonal regeneration.