Immunocytochemical studies in the past, using alternate serial sections to localize individual antigens, concluded that there was no close relationship between renin- and kallikrein-containing structures in the rat kidney. We have investigated this relationship by simultaneously localizing renin and kallikrein in the same section using immunoperoxidase with two different chromogens. Analysis of serial kidney sections from three rats indicated that kallikrein-containing late distal tubular cells corresponded in their distribution to connecting tubule cells. They were observed in the proximity (less than 3 micrograms) of renin-containing JG cells in 66.6% of the superficial (N = 30), 46.6% of the midcortical (N = 15) and 26.7% of the juxtamedullary (N = 15) afferent arterioles surveyed. When traced through serial sections, 90% of the afferent arterioles from superficial glomeruli (N = 30), 86.7% of the afferent arterioles from midcortical glomeruli (N = 15) and 73.3% of those from juxtamedullary glomeruli (N = 15) came within 3 micrograms of a late distal tubule showing some kallikrein-positive cells. These cells were adjacent to the afferent arteriole in 67 to 80% of the arterioles surveyed. This spatial relationship suggests an anatomical basis for a possible interaction between the afferent arteriole, containing renin-positive JG cells, and kallikrein-positive late distal tubular cells.