The activity of NADPH-diaphorase (NADPH-d), a marker for nitric oxide synthase (NOS), was examined histochemically in the rat and mice retina. Mice in which the neuronal NOS gene has been disrupted (nNOS- mice) were used for specificity controls. Light microscopically a few amacrine cells were heavily stained. Other cells were stained weakly or not at all. Under the electron microscope, formazan precipitates were detectable on membranes of endoplasmic reticulum, nuclear envelope, mitochondria, and, in a few cases, the Golgi complex. Bipolar, horizontal, and Müller cells, were if at all, sparsely labeled with formazan. Labeled mitochondria were observed in rod endings and in inner segments of photoreceptors. Outer segments of photoreceptors and ganglion cells were completely free of reaction product. The NADPH-d reaction in wild-type mice displayed a similar distribution pattern to that in rats. Retinae of nNOS- mice showed a complete lack of prominent NADPH-d stained (amacrine) cells. None or a very few labeled membranes were seen.