Nongenomic actions of thyroxine modulate intermediate filament phosphorylation in cerebral cortex of rats.

The developmental effects of thyroid hormones (TH) in mammalian brain are mainly mediated by nuclear receptors regulating gene expression. However, there are increasing evidences of nongenomic mechanisms of these hormones associated with kinase- and calcium-activated signaling pathways. In this context, the aim of the present work was to investigate the signaling pathways involved in the mechanism of action of TH on cytoskeletal phosphorylation in cerebral cortex of 15-day-old male rats. Results showed that L-thyroxine (L-T4) increased the intermediate filament (IF) phosphorylation independently of protein synthesis, without altering the total immunocontent of these proteins. Otherwise, neither 3,5,3'-triiodo-L-thyronine (L-T3) nor neurotransmitters (GABA, ATP, L-glutamate or epinephrine) acted on the IF-associated phosphorylation level. We also demonstrated that the mechanisms underlying the L-T4 effect on the cytoskeleton involve membrane initiated actions through Gi protein-coupled receptor. This evidence was reinforced by the inhibition of cyclic adenosine 5'-monophosphate (cAMP) levels. Moreover, we showed the participation of phospholipase C, protein kinase C, mitogen-activated protein kinase, calcium/calmodulin-dependent protein kinase II, intra- and extracellular Ca2+ mediating the effects of L-T4 on the cytoskeleton. Stimulation of 45Ca2+ uptake by L-T4 was also demonstrated. These findings demonstrate that L-T4 has important physiological roles modulating the cytoskeleton of neural cells during development.

Rapid responses to thyroxine in the testis: active protein synthesis-independent pathway.

We investigated the involvement of protein synthesis in the stimulatory action of thyroid hormones on amino acid accumulation and characterized K(+) currents involved in the hyperpolarizing effect of thyroxine (T(4)) on Sertoli cells. Immature rat testes were incubated in Krebs Ringer-bicarbonate buffer (KRb) in the presence of [(14)C]methylaminoisobutyric acid with and without T(4), 3,5,3'-l-triiodothyronine (T(3)) and/or cycloheximide. Sertoli cells were monitored by intracellular recording in a chamber perfused with KRb with and without T(4), T(3) and/or blockers, and the membrane potential was monitored. T(4) and T(3) stimulated amino acid accumulation and protein synthesis. Treatment with cycloheximide diminished T(3) stimulatory actions on amino acid accumulation but had no effect on T(4) action. Both hormones elicited a hyperpolarization of the Sertoli cell membrane potential which involved K(+) channels, since TEA and apamin abolished this effect. These findings on rapid membrane actions of thyroid hormone in the testis suggest that some effects of T(4) are modulated by non-genomic mechanisms.

Effect of 3,5,3'-triiodo-L-thyronine on amino acid accumulation and membrane potential in Sertoli cells of the rat testis.

The purpose of this study was to investigate the effect of T3 on amino acid accumulation and on the membrane potential of Sertoli cells of immature rat testes. Testes of pre-pubertal and pubertal rats were pre-incubated (30 min) in Krebs-Ringer bicarbonate buffer and incubated in the presence of [14C]methylaminoisobutyric acid with and without T3 for 15, 45 and 60 min. The hormone (10(-6) M and 10(-7) M) significantly stimulated amino acid accumulation in 6 and 13-day old rat testes but did not have any effect in neonatal and pubertal animals. T3 produced a dose-dependent hyperpolarizing effect at concentrations of 10(-6) M, 10(-5) M, 2 x 10(-5) M and 10(-4) M. We conclude that T3 induces a membrane hyperpolarization in Sertoli cells and stimulates amino acid accumulation in immature rat testes, demonstrating that the hormone has a rapid plasma membrane action.