New perspectives in melatonin uses.

This review summarizes the metabolism, secretion, regulation and sites of action of melatonin. An updated description of the melatonin receptors, including their signal transduction mechanisms, distribution and characterization of receptor genes, is given. Special emphasis is focused on the clinical aspects and potential uses of melatonin in the sleep-wake rhythms, in the immune function, in cancer therapy, in neuroprotection against oxidative damage and antioxidant activities in different tissues. Finally, combined effects of melatonin with other drugs are discussed.

Effect of retinal ablation on the expression of calbindin D28k and GABA in the chick optic tectum.

The effect of retinal ablation on qualitative and quantitative changes of calbindin D28k and GABA expression in the contralateral optic tectum was studied in young chicks. Fifteen days old chicks had unilateral retinal ablation and after 7 or 15 days, calbindin expression was analyzed by Western blot and immunocytochemistry. Neuronal degeneration was followed by the amino-cupric silver technique. After 15 days, retinal lesions produced a significant decrease in calbindin immunostaining in the neuropil of layers 5-6 and in the somata of neurons from the layers 8 and 10 of the contralateral tectum, being this effect less marked at 7 days post-lesion. Double staining revealed that 50-60% of cells in the layers 8 and 10 were calbindin and GABA positive, 30-45% were only calbindin positive and 5-10% were only GABAergic neurons. Retinal ablation also produced a decrease in the GABA expression at either 7 or 15 days after surgery. At 7 days, dense silver staining was observed in the layers 5-6 from the optic tectum contralateral to the retinal ablation, which mainly represented neuropil that would come from processes of retinal ganglion cells. Tectal neuronal bodies were not stained with silver, although some neurons were surrounded by coarse granular silver deposits. In conclusion, most of calbindin molecules are present in neurons of the tectal GABAergic inhibitory circuitry, whose functioning apparently depends on the integrity of the visual input. A possible role of calbindin in the control of intracellular Ca2+ in neurons of this circuit when the visual transmission arrives to the optic tectum remains to be studied.

DL-Buthionine-S,R-sulfoximine affects intestinal alkaline phosphatase activity.

The susceptibility of intestinal alkaline phosphatase to DL-buthionine-S,R-sulfoximine was investigated in chicks fed a commercial diet. The results show that DL-buthionine-S,R-sulfoximine produced inhibition of intestinal alkaline phosphatase activity. This effect showed dose- and time-dependency and it was caused by either in vivo DL-buthionine-S,R- sulfoximine administration or in vitro DL-buthionine-S,R-sulfoximine incubation with villus tip enterocytes. DL-Buthionine-S,R-sulfoximine did not act directly on intestinal alkaline phosphatase but it provoked glutathione depletion which led to changes in the redox state of the enterocyte as shown by the production of free hydroxyl radicals and an incremental increase in the carbonyl content of proteins. The reversibility of the buthionine sulfoximine effect on intestinal alkaline phosphatase was proved by addition of glutathione monoester to the duodenal loop.

Vitamin D affects Krebs cycle NAD-linked oxidoreductases from chick intestinal mucosa.

Vitamin D3 administration affects the NAD-linked oxidoreductase activities of Krebs cycle from intestinal mucosa of vitamin D-deficient chicks. Vmax values were increased in all of them, while K0.5 for substrate remained unchanged except for 2-oxoglutarate dehydrogenase, which showed lower affinity for oxoglutarate. Addition of Ca2+ to the incubation medium increased the affinity of 2-oxoglutarate dehydrogenase and NAD-isocitrate dehydrogenase for their substrates either in the vitamin D3 treated group or in the control one. The activity of succinate dehydrogenase, a FMN-dependent oxidoreductase, was not modified by vitamin D3 administration. The oxygen consumption of the intestinal mitochondria was not altered by cholecalciferol treatment to vitamin D-deficient chicks. The reason why vitamin D3 selectively affects the NAD-linked oxidoreductase activities of the Krebs cycle remains unknown. The vitamin D hormone, 1,25(OH)2D3, appears to be the mediator of the response.