The insulin family of peptides and their receptors influence cellular growth in very early preimplantation embryos. In this study their expression and role in renal organogenesis was investigated. By immunofluorescence microscopy and in situ hybridization, insulin receptor (IR) expression was seen in the ureteric bud branches and early nephron precursors in mouse metanephroi harvested at day 13 of gestation. The expression gradually decreased in successive stages of gestation, and it was confined mainly to renal tubules in 1-week-old mice. Similar developmental regulation of the IR and insulin was observed by reverse transcriptase-polymerase chain reaction (RT-PCR) analyses. Addition of insulin into the culture medium at low concentrations, ranging from 40 to 400 ng͞ml, induced trophic changes and increased [ 3 H]thymidine incorporation in the embryonic renal explants, and inclusion of IR -subunitspecific antisense oligodeoxynucleotide caused marked dysmorphogenesis and growth retardation of the metanephroi. Specificity of the antisense effect was reflected by immunoprecipitation experiments in which translational blockade of the  subunit of the IR was observed. RT-PCR analyses revealed that the ␣ subunit of the IR was unaffected by the antisense treatment of metanephric explants. Concomitantly, de novo synthesis of morphogenetic regulatory extracellular matrix proteins, especially the proteoglycans, was decreased. Gel-shift analyses indicated a failure in the activation of c-fos promoter region binding protein(s) by insulin in the antisense oligodeoxynucleotidetreated explants. These studies suggest that insulin and its putative receptor are developmentally regulated in the murine embryonic metanephros, and they play a role in renal organogenesis, possibly by affecting other modulators of morphogenesis-i.e., extracellular matrix proteins and protooncogenes.Organogenesis in embryonic life constitutes a series of intricate processes which involve differentiation and rapid proliferation of pluripotent cells under the influence of a number of growth factors, including insulin and insulin-like growth factors (IGFs) (1-3). The IGFs and several other growth factors, upon binding to their putative transmembrane receptors, induce a multitude of effects-e.g., cell proliferation and induction of immediate-early genes (IEGs), and thus play a vital role in the development of various organs (3-5). However, the role of insulin and its receptor in embryonic development has received limited attention, which in part may be related to the fact that insulin and its receptor are primarily known for their regulation of glucose metabolism rather than as growth modulators (6). Nonetheless, the binding of insulin to the preimplantation mouse embryos, and expression of insulin receptor (IR) in the prepancreatic͞precardiac stage of avian embryos and in early mid-gestational human fetuses raises the possibility for the growth potential of insulin (7-10). Furthermore, the expression of insulin during the prepancreatic stage at si...