Microtubule dynamics regulates the level of endothelin-B receptor in rat cultured astrocytes.

Abstract

We investigated the effect of cytoskeleton modulators on endothelin-B (ET(B)) receptor expression in rat primary cultured astrocytes. Northern blot analysis and a binding study revealed that colchicine and nocodazole, microtubule-disrupting agents, decreased the levels of both ET(B) receptor mRNA and the number of ET-1 binding sites in quiescent astrocytes. Down-regulation of both ET(B) receptor mRNA and the number of binding sites for ET-1 was also observed in quiescent astrocytes treated with taxol, a microtubule-stabilizing agent. In contrast, neither beta-lumicolchicine, an inactive isomer of colchicine, nor cytochalasin D, a microfilament-disrupting agent, influenced ET(B) receptor expression. The level of ET(B) receptors in astrocytes was affected by the cell state, namely, proliferative, quiescent, or differentiated state. The order of ET(B) receptor expression according to the cell state was proliferative state < quiescent state << differentiated state induced by dibutyryl cyclic AMP. Also, in proliferative astrocytes and differentiated astrocytes, colchicine significantly down-regulated both ET(B) receptor mRNA and the number of binding sites for ET-1. However, thymidine assay revealed that colchicine did not change quiescent astrocytes and differentiated astrocytes to a proliferative state. Furthermore, the increase in glutamine synthetase activity in differentiated astrocytes was not affected by colchicine. These results suggest that microtubule dynamics possibly regulates ET(B) receptor expression in astrocytes without affecting the cell state.