Neurotensin and neuromedin N elevate the cytosolic calcium concentration via transiently appearing neurotensin binding sites in cultured rat cortex cells

Abstract

Through assessment of the changes in the intracellular free-calcium concentration ([Ca 2+] i), which was measured using the calcium sensitive dye, fura-2, the character of the neurotensin (NT) binding sites which appeared transiently during the early ontogenetic stage in the rat cerebral cortex was analyzed in primary cultures of cerebral cortex cells from neonatal rats. NT (1–1000 nM) elevated [Ca 2+] i of the cells even when extracellular calcium was chelated with 1 mM ethylene glycol-bis(β-aminoethylether)- N, N, N′, N′-tetraacetic acid (EGTA). These findings suggest that the transiently appearing NT-binding sites in the cortex are receptors for NT and that some of the NT-induced increase in [Ca 2+] i is due to mobilization from the intracellular calcium store. Further application of NT after 10 min washing caused an increase in [Ca 2+] i again. This is in contrast to the findings for cortical slices from adult rats and mRNA-injected oocytes; desensitization due to NT was of long duration and further application of NT failed to activate the neurons which had responded the first time to NT. These facts suggest that the character of the NT-binding sites in the cerebral cortex differs between neonatal and adult rats. In addition, we showed that neuromedin N had a similar property to NT as to mobilization of [Ca 2+] i and acted only on NT-responsive cells, suggesting the interaction between NT and neuromedin N at the postsynaptic level via the same receptor.