In this study, we investigated the tissue distribution and expression of monocarboxylate transporter 1 (MCT1) along the gastrointestinal tract of sheep. Western blot analysis suggested the presence of MCT1 as a 43-kDa protein in immunoblots of membranes from the various tissues examined. The results of Western blotting were further confirmed by immunohistochemical studies, which revealed intense immunoreactivity for the MCT1 protein in the forestomach (rumen, reticulum and omasum) and large intestine (caecum, proximal and distal colon). Moderate reactivity, however, was detected in the abomasum, while no immunoreactivity could be seen in any regions of the small intestine examined. Furthermore, MCT1 was expressed at the mRNA level as determined by reverse transcriptase polymerase chain reaction (RT-PCR), which showed a band of the expected size (300 bp) in all tissues examined. From these results we concluded that MCT1 protein is highly expressed and distributed in the stomach and large intestine of sheep suggesting that MCT1 may play a significant role in the transport of short chain fatty acids and their metabolites in the gastrointestinal tract of ruminants.