Regulation of cytoskeletal proteins by thyroid hormone during neuronal maturation and differentiation.

Primary cultures of neurons from 16- to 17-day-old embryonic rat cerebra were maintained for 3 weeks in thyroid hormone deficient (THdef) and thyroid hormone supplemented (THsup) media to investigate how TH regulates the cytoskeletal (CSK) proteins during neuronal differentiation and maturation. Two distinct phases of regulation of triton-insoluble CSK-proteins by TH were discernible--an early phase (days 4-8 of culture) when TH-deficiency resulted in down-regulation of these proteins and a late phase (days 16-20) involving up-regulation of these proteins. In contrast, the triton-soluble non-CSK proteins always remained up-regulated by TH. The two main effects of TH-deficiency were retarded neurite outgrowth and altered neuronal morphology. Of all the CSK proteins, actin was found to be predominantly sensitive. Alterations in the level of CSK actin during neuronal maturation were found to be parallel to changes in steady-state level of actin mRNA as well as actin synthesis. However, these TH-induced changes (up-regulation of actin during the early phase and down-regulation during the late phase) did not lead to parallel changes in the level of soluble G-actin which was comparable at both days 8 and 16 in THdef and THsup cultures. Quantitation of different forms of intracellular actin revealed that G-actin level declined by about 50% between days 8 and 16. In the case of THsup neurons, this reduction in G-actin was accompanied by a parallel increase in the non-CSK F-actin, whereas TH-deficiency resulted in a corresponding increase in CSK F-actin during the terminal differentiation of neurons. Thus TH regulates the biogenesis of CSK-proteins with a predominant effect on actin and the transformation of G-actin into non-CSK F-actin appears to be the key step in neuronal maturation which is affected by hypothyroidism.