Creatine and phosphocreatine are required to maintain ATP needed for normal retinal function and development. The aim of the present study was to determine the distribution of the creatine transporter (CRT) to gain insight to how creatine is transported into the retina. An affinity-purified antibody raised against the CRT was applied to adult vertebrate retinas and to mouse retina during development. Confocal microscopy was used to identify the localization pattern as well as co-localization patterns with a range of retinal neurochemical markers. Strong labelling of the CRT was seen in the photoreceptor inner segments in all species studied and labelling of a variety of inner neuronal cells (amacrine, bipolar and ganglion cells), the retinal nerve fibers and sites of creatine transport into the retina (retinal pigment epithelium, inner retinal blood vessels and perivascular astrocytes). The CRT was not expressed in Muller cells of any of the species studied. The lack of labelling of Muller cells suggests that neurons are independent of this glial cell in accumulating creatine. During mouse retinal development, expression of the creatine transporter progressively increased throughout the retina until approximately post-natal-day 10, with a subsequent decrease. Comparison of the distribution patterns of the CRT in vascular and avascular vertebrate retinas, and studies of the mouse retina during development indicate that creatine and phosphocreatine are important for ATP homeostasis.