Visualisation of somatostatin receptor sst(3) in the rat central nervous system

Somatostatin actions are mediated through G-protein coupled receptors named sst(1) to sst(5). We used an affinity-purified polyclonal antibody AS-69, directed against a specific N-terminal peptide sequence of sst(3) to determine the immunohistochemical distribution of the sst(3) receptor in the rat and human brain. The specificity of the antibody was shown by Western blotting experiments using an N-terminal sst(3) fusion protein. Enzymatic deglycosylation experiments were combined to blotting experiments on a sst(3)-transfected cell line and rat brain membrane proteins and with immunocytochemistry on the sst(3)-transfected cell line. These studies showed that the antibody detected the deglycosylated sst(3) receptor protein. Immunohistochemical staining showed that sst(3) immunoreactivity recognised by this N-terminal antiserum was widely distributed throughout the brain with cells and processes labelled in the cerebral cortex, regions of the limbic system (including the hippocampal formation, some amygdaloid regions, some basal ganglia nuclei and regions from the nucleus basalis complex), the habenula, the hypothalamus, the thalamus, different mesencephalic structures (substantia nigra, zona incerta, superior colliculus), the reticular formation, the cerebellum. The distribution of immunoreactivity was in good general agreement with that predicted from the localisation of sst(3) mRNA and radio-ligand binding studies; however, due to the preference of AS-69 towards the deglycosylated receptor, it appears that the sst(3) immunoreactivity detected may correspond largely to the deglycosylated receptor. This study on the immunohistochemical distribution of the sst(3) receptor in the brain may provide a better understanding of the central actions of somatotropin release-inhibiting factor (SRIF).