The metabotropic glutamate receptor subtypes mGluR2 and mGluR5, which are thought to be coupled respectively to the inhibitory cyclic adenosine monophosphate (cAMP) cascade and the phosphatidylinositol hydrolysis/Ca2+ cascade, are known to be expressed on Golgi cells in the granular layer of the rat cerebellar cortex. In the present immunohistochemical study with a monoclonal antibody against mGluR2 and a polyclonal antibody for mGluR5, we examined whether or not mGluR2- and mGluR5-like immunoreactivities were both present in single Golgi cells in the rat cerebellar cortex. In double immunofluorescence histochemistry, no Golgi cells showed mGluR2- and mGluR5-like immunoreactivities simultaneously. Of the total number of Golgi cells immunoreactive for mGluR2 or mGluR5, about 90% were mGluR2-like immunoreactive, and about 10% were mGluR5-like immunoreactive. Golgi cells with mGluR2-like immunoreactivity were distributed evenly in the granular layer of all the cerebellar regions, while those with mGluR5-like immunoreactivity were distributed more frequently in the I, II, VII-X lobules of the vermis and the copula pyramidis of the hemisphere than in other cerebellar regions. The results indicate that Golgi cells containing mGluR2 are segregated from those possessing mGluR5. These two populations of Golgi cells, each equipped with a different metabolic glutamate receptor coupled to a different intracellular signal transduction system, may play different roles in the glutamatergic neuronal circuits in the cerebellar cortex.