Insulin Release and the Microtubular System of the Islets of Langerhans: Effects of Insulin Secretagogues on Microtubule Subunit Pool Size

Abstract

An assay system was devised to estimate the pool size of microtubule subunits in islet cells, and to study the importance of the equilibrium between polymerised microtubules and their subunits in the regulation of insulin release. The assay was based on the observation that colchicine binds specifically and quantitatively to microtubule protein subunits, but not to intact microtubules.

Vinblastine and cold treatment, which have been shown to cause disaggregation of microtubules into subunits and to inhibit insulin release from islets, increased the number of colchicine binding subunits. D₂O, which promotes stability of microtubules and increases their number in islet cells, caused a decrease in subunit concentration. These results suggest that changes in the equilibrium between polymerised microtubules and their subunits could be studied by measuring the size of the subunit pool.

When insulin release was stimulated, by incubating islets in high glucose concentrations or by increasing the intracellular concentration of cyclic AMP there was a reduction in the content of subunit protein. Conversely, when insulin release was inhibited by removal of calcium from the incubation medium there was a shift in the microtubule subunit equilibrium to give an increase in the number of subunits assayed. These results indicate that changes in the equilibrium between subunits and microtubules may play an important role in regulating rates of insulin secretion.

Key words

Islets of Langerhans - Colchicine Binding Assay - Vinblastine - D₂O - Microtubule Subunit Equilibrium - Glucose - Cyclic AMP - Calcium

1 Present address: Department of Biochemistry, The University of Leicester, Leicester LE 1 7RH, U.K.