The distribution of cathepsin D (CD) was surveyed in rat tissues by light microscopic immunocytochemistry. Although immunoreactive CD was detected in all tissues examined, there was a marked difference in the amount in the cytoplasm of different cell types of the same organ. In the retina large amounts of CD were present in the pigment epithelium, ganglion cells, and Müller cells. Moderate to large amounts of CD were also found in neuronal perikarya of the gastrointestinal tract and adrenal medulla; in macrophages in the lung, liver, and spleen; in some secretory cells of the submandibular and lacrimal glands; in parts of renal distal convoluted and collecting tubules; and in the surface transitional epithelium of the calyx, ureter, and urinary bladder. Other cells adjacent to cells containing large amounts of the enzyme had little or no detectable CD themselves. These included hepatocytes, the proximal tubular cells of the kidney, selected cells of the submandibular gland, cells of the zona glomerulosa of the adrenal cortex, and lymphocytes in lymphoid organs. The localization of CD indicates that its degradative effect is exerted preferentially in certain cell types and suggests that physiological influences on CD may have a variety of effects in different organs.