Inhibition of glycogen synthase kinase 3beta activity regulates proliferation of cultured cerebellar granule cells.

Insulin-like growth factor I (IGF-I) is mitogenic for several types of neuronal progenitors including cerebellar granule neuron progenitors. The present study confirms that IGF-I can function as a mitogen in purified cultures of cerebellar granule cells and identifies intracellular signal transduction molecules that mediate this mitogenesis. In cultured granule cells, IGF-I inhibits GSK-3 activity and leads to phosphorylation of serine9 an inhibitory site on GSK-3beta. Phosphoinositide 3-kinase (PI3-K) activation by IGF-I can lead to phosphorylation and inactivation of GSK-3. A PI3-K inhibitor, LY294002, completely inhibited IGF-I-induced proliferation with half-maximal inhibition occurring at a concentration (1.5 micrograms) close to its reported IC50 value for inhibition of PI3-K. Lithium chloride (LiCl), a direct inhibitor of GSK-3beta, can alone stimulate granule cell proliferation and enhance proliferation induced by IGF-I. LiCl can reverse the inhibitory effect of LY294002 on granule cell proliferation suggesting that GSK-3 inhibition may be downstream of PI3-K activation in IGF-I's mitogenic pathway. Experiments further show that the expression of a dominant active form of GSK-3beta antagonizes IGF-I-induced mitogenesis. These studies support a role for inhibition of GSK-3beta activity in the signal transduction pathway by which IGF-I regulates granule neuron progenitor proliferation.