Connective tissue growth factor (CTGF) is a 38 kDa modular protein that is involved in processes such as cell proliferation, survival, migration, adhesion and extracellular matrix production. Target cells for CTGF include fibroblasts, smooth muscle cells and endothelial cells. Using a specific peptide antibody, CTGF was localized in tissue sections obtained from mouse embryos between days 7.5 and 14.5 of gestation. On day 7.5, CTGF was present at high levels in the embryonic ectoderm, mesoderm, and chorion, with weaker levels in the squamous endoderm. No CTGF was detected in Reichert's membrane, parietal endoderm, and visceral endoderm. Decidual cells of the maternal uterus stained strongly for CTGF. There was no specific staining for CTGF on day 11.5 but by day 13.5, the protein was detectable in a limited number of structures, most notably the liver, thymus, lung, and intestine. By day 14.5, staining for CTGF had become extensive and was present at high or moderate levels in the thymus, aorta, trachea, liver, lung, adrenal gland, kidney, iliac sinus, olfactory region, tongue, pharynx, esophagus, thyroglossal duct, choroid plexi, Rathke's pouch, stomach, pancreas, and midgut. Relatively weak staining was present in the heart and dorsal root ganglia. There was no staining in the vitelline duct or in the outer cortical or medullary regions of the brain. Among specific cell types, CTGF was localized at high levels in secretory and absorptive epithelial cells and liver parenchyma cells, at moderate levels in vascular endothelial cells and myoblasts and at low levels in mesenchymal and connective tissue cells. These data show that various tissues and organ systems produce CTGF in a specific temporo-spatial pattern during embryogenesis and support a role for CTGF in cellular differentiation and development during prenatal life.