Two complementary DNAs for the organic anion transporter subtypes oatp2 and oatp3, which transport thyroid hormones as well as taurocholate, were isolated from a rat retina cDNA library. The sequence of oatp2 is identical to that recently reported (Noé , B., Hagenbuch, B., Stieger, B., and Meier, P. J. (1997) Proc. Natl. Acad. Sci. U. S. A. 94, 10346 -10350), whereas the sequence of oatp3 is novel. oatp3 consists of 670 amino acid residues and exhibits a structural architecture common to the organic anion transporter family, possessing the 12 putative membrane-spanning segments. Oocytes injected with oatp2 and oatp3 cRNAs showed taurocholate uptake in a saturable manner. The oatp2 and oatp3 cRNAinjected oocytes also showed significant uptake of both thyroxine and triiodothyronine. Northern blot and in situ analyses showed that the oatp2 mRNA was widely expressed in neuronal cells of the central nervous system, especially in the hippocampus, cerebellum, and choroid plexus as well as in the retina and liver. The oatp3 mRNA was highly expressed in the kidney and moderately abundant in the retina. This suggests that oatp2 and oatp3 are multifunctional transporters involved in the transport of thyroid hormones in the brain, retina, liver, and kidney.A homeostatic system controls the fluid environment in the brain and keeps its chemical composition relatively constant compared with that of plasma. One mechanism is the bloodbrain barrier, which selectively transports chemical substances via capillary endothelial cells (1). A second essential component is the choroid plexus (blood-cerebrospinal fluid barrier), which secretes or takes up specific chemical substances (2). Although the presence of specific transporting mechanisms has long been postulated, little is known about their molecular identity. Recent molecular biological studies revealed the organic anion transporter family: the Na ϩ -independent organic anion-transporting polypeptide oatp1 from rat liver, which transports bile acid, bromosulfophthalein (BSP), 1 and conjugated and unconjugated steroid hormones (3, 4); the kidney-specific transporter OAT-K1, which transports methotrexate in the basolateral membrane of renal tubules (5); and the prostaglandin transporter (6). Moreover, physiological studies have suggested the presence of other members of the organic anion transporter family (7). Noé et al. (8) have recently reported that a new organic anion transporter subtype (oatp2) is present in rat brain and liver and that the oatp2-expressed oocytes transported cardiac glycoside as well as taurocholate. However, the endogenous substrate of oatp2 and the regional distribution in the brain have not been revealed.It has been suggested that thyroid hormones are transported into the brain via the blood-brain barrier (9) or via the choroid plexus (10). To reveal this mechanism, we focused on the retina. In the retina, the retinal pigment epithelium is the unique source of transthyretin synthesis, and it serves to transport thyroxine (T4) across the blood-retina barr...