[Show abstract][Hide abstract] ABSTRACT: Serotonin (5-hydroxytryptamine, 5-HT), a precursor for melatonin production, is produced abundantly in the pineal gland of all vertebrate animals. The synthesis of 5-HT in the pineal gland is rate limited by tryptophan hydroxylase 1 (TPH1) whose activity displays a twofold increase at night. Earlier studies from our laboratory demonstrate that pineal 5-HT secretion exhibits dynamic circadian rhythms with elevated levels during the early night, and that the increase is controlled by adrenergic signaling at night. In this study, we report that (a) 5-HT total output from the pineal gland and TPH1 protein levels both display diurnal rhythms with a twofold increase at night; (b) stimulation of cAMP signaling elevates 5-HT output in vivo; (c) 5-HT total output and TPH1 protein content in rat pineal gland are both acutely inhibited by light exposure at night. Consistent with these findings, molecular analysis of TPH1 protein revealed that (a) TPH1 is phosphorylated at the serine 58 in vitro and in the night pineal gland; and (b) phosphorylation of TPH1 at this residue is required for cAMP-enhanced TPH1 protein stability. These data support the model that increased nocturnal 5-HT synthesis in the pineal gland is mediated by the phosphorylation of TPH1 at the serine 58, which elevates the TPH1 protein content and activity at night.
Journal of Pineal Research 09/2008; 45(4):506-14. DOI:10.1111/j.1600-079X.2008.00627.x · 9.60 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: This study describes the development of a new technique for long-term measurement of daily 5-hydroxytryptamine (5-HT) and melatonin contents in the pineal gland of freely moving rats. The technique features a number of novel improvements over previous protocols. It allows visualization of the pineal gland for accurate targeting of the guide cannula, which minimizes bleeding; incurs no direct injury to the surrounding brain tissues; and causes no interference with the sympathetic innervation from the superior cervical ganglia. Robust releases of melatonin and indole precursors were continuously monitored quantitatively and reproducibly for more than 2 wk in the same animal. In addition, effects of pharmacological agents on in vivo pineal circadian rhythms can be studied reproducibly over time, and gene expression profiles can be correlated with physiological consequences in single animals. Using these approaches, it is found that beta-adrenergic activation leads to decreased release of 5-HT, and that increased cAMP signaling in vivo results in activation of N-acetyltransferase gene induction and melatonin production. These studies will enhance the understanding of signaling pathways that regulate pineal 5-HT and melatonin synthesis and secretion.
Journal of Pineal Research 10/2003; 35(2):118-24. DOI:10.1034/j.1600-079X.2003.00064.x · 9.60 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The 3-O-sulfotransferases (3OSTs) catalyze the addition of sulfate groups at the 3-OH site of glucosamine in heparan sulfate proteoglycans, which serve as critical mediators of various biological functions. We demonstrate that the 3OST2 isoform is expressed at high levels in the rat pineal specifically during the daylight hours. The dramatic diurnal rhythm of 3OST2 is regulated by central clock-controlled activities of the superior cervical ganglion, persists in constant darkness, and is inducible by light at nighttime. Importantly, 3OST2 transcription is blocked by beta-adrenergic agonists that activate the pineal melatonin formation and is induced by beta-adrenergic antagonists, which block melatonin production in vivo. Because of the inverse expression and regulation patterns of 3OST2 with serotonin N-acetyltransferase, the enzyme controlling the melatonin rhythm in the pineal, we tested the effects of forced expression of 3OST2 in the night pineals on N-acetyltransferase gene expression and melatonin production and found that, surprisingly, 3OST2 expression at night fails to interfere with melatonin synthesis. These data suggest 3OST2 may serve a unique function in the pineal that may be independent of melatonin formation.
[Show abstract][Hide abstract] ABSTRACT: Using on-line microdialysis, we have characterized in vivo dynamics of pineal 5-hydroxytryptamine (5-HT; serotonin) release. Daily pineal 5-HT output is triphasic: (i) 5-HT levels are constant and high during the day; (ii) early in the night, there is a novel sharp rise in 5-HT synthesis and release, which precedes the nocturnal rise in melatonin synthesis; and (iii) late in the night, levels are low. This triphasic 5-HT production persists in constant darkness and is influenced strongly by intrusion of light at night. We demonstrate that both diurnal 5-HT synthesis and 5-HT release are activated by sympathetic innervation from the superior cervical ganglion and show that these processes are controlled by distinct receptors. The increase in 5-HT synthesis is controlled by beta-adrenergic receptors, whereas the increase in 5-HT release is mediated by alpha-adrenergic signaling. On the other hand, the marked decrease in 5-HT content and release late at night is a passive process, influenced by the extent of melatonin synthesis. In the absence of melatonin synthesis, the late-night decline in 5-HT release is prevented, reaching levels roughly twice as high as that of the day value. In summary, our results demonstrate that 5-HT levels display marked circadian rhythms that depend on adrenergic signaling.
Proceedings of the National Academy of Sciences 05/2002; 99(7):4686-91. DOI:10.1073/pnas.062585499 · 9.67 Impact Factor