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S W Ying

Publications and source records attributed to S W Ying.

13 recordsLinked to original sources
PubMed1998

Chronic exposure to melatonin receptor agonists does not alter their effects on suprachiasmatic nucleus neurons.

Previous studies have demonstrated that melatonin and a novel melatonin receptor agonist, S20098 (N-[2-(7-methoxy-1-naphthyl) ethyl] acetamide), regulate neuronal firing activity of photically responsive cells in the suprachiasmatic nucleus in vivo. In the present study, we used several different methods to investigate the effects of chronic daily treatment with melatonin, S20098 (1.0 mg/kg, s.c.) or control vehicle for 14 d on responsiveness of suprachiasmatic nucleus cells to these agonists. Both chronic and acute application of drugs were carried out during the day-night transition period. We confirmed that suprachiasmatic nucleus cells from control animals were most sensitive at this circadian phase. Chronic drug treatments did not alter sensitivity of photically responsive suprachiasmatic nucleus cells to S20098 or melatonin given intraperitoneally (i.p.) or iontophoretically in vivo. Suprachiasmatic nucleus cells studied in brain slice preparations also responded similarly to micropressure ejections of melatonin receptor agonists regardless of drug pretreatment. These results indicate that chronic melatonin receptor agonist pretreatment does not result in desensitization of suprachiasmatic nucleus neuronal responses to these agonists during the daily phase of maximum melatonin sensitivity.

Acetamides↗
PubMed1997

5-HT7 receptors mediate serotonergic effects on light-sensitive suprachiasmatic nucleus neurons.

Serotonin (5-HT) has been shown to phase shift circadian rhythms in mammals and to affect responses of the circadian system to light, but it is not clear which receptors are involved in these actions. We found that drugs which act as 5-HT1A receptor agonists suppressed photic responses of hamster SCN cells, but these drugs also exhibit high affinity for the recently cloned 5-HT7 receptor. We therefore studied the effects of 5-HT agonists and antagonists with differential affinities for 5-HT7 and 5-HT1A receptors on responses of hamster SCN cells to retinal illumination. We confirmed that the 5-HT receptor agonists 5-HT, 8-OH-DPAT and 5-CT, dose-dependently reduced photic activation of SCN cells. These effects could be blocked by co-application of antagonists with high affinities for 5-HT7 receptors: ritanserin or clozapine. The 5-HT1A/B/D antagonist, cyanopindolol, which is inactive at 5-HT7 receptors, did not antagonize the actions of 8-OH-DPAT. Selective 5-HT1A antagonists, WAY100635 and p-MPPI, had weak or no antagonist effects on the responses to 8-OH-DPAT in the SCN, but they effectively antagonized the actions of 8-OH-DPAT in the hippocampus. In the cerebellar cortex where few 5-HT7 receptors are present, ritanserin failed to antagonize the effects of 8-OH-DPAT. Our results indicate that the 5-HT7 receptor subtype plays a major role in mediating the effects of 5-HT on photic responses of SCN cells in the hamster.

Animals↗
PubMed1997

Cloning and characterization of the ori region of pSW1200 of Erwinia stewartii: similarity with plasmid P1.

The ori region of an Erwinia stewartii plasmid, pSW1200 (106 kb), has been cloned and sequenced. This region consists of a gene encoding a protein which has 91% similarity and 73% identity with the RepA protein of bacteriophage P1. The ori region also consists of eight copies of 19-bp iterons which are highly homologous to the iterons of P1. Similar to plasmid P1, pSW1200 replicon has a copy number of approximately 1. On the other hand, the copy number increases about ninefold if three of the iterons located downstream from repA gene are deleted. We also demonstrate that pGEM-5Z consisting of a copy of P1 iteron is incompatible with a pSW1200 derivative, pSW1201, suggesting that pSW1200 and P1 DNA are incompatible and both belong to the IncY group.

Base Sequence↗
PubMed1996

Melatonin analogues as agonists and antagonists in the circadian system and other brain areas.

We studied the effects of drugs related to melatonin on neuronal firing activity in the suprachiasmatic nucleus, intergeniculate leaflet and other brain areas in urethane-anesthetized Syrian hamsters. We tested melatonin and two naphthalenic derivatives of melatonin, a putative agonist (S20098: N-[2-(7-methoxy-1-naphthyl)ethyl]acetamide), and a putative antagonist (S20928: N-[2-(1-naphthyl)ethyl]cyclobutyl carboxamide). Both melatonin and S20098 given intraperitoneally (i.p.) were able to suppress firing rates of cells in a similar dose-dependent manner, but the effects of S20098 were longer lasting. Iontophoresis of melatonin dose dependently depressed spontaneous and light-evoked activity of cells in the suprachiasmatic nucleus and intergeniculate leaflet, while iontophoresis of S20098 was relatively ineffective, probably because it is a poorly charged compound. S20928 (2.0-10 mg/kg, i.p.) alone decreased firing rates of light-sensitive cells by 25-50% for 5-30 min in the suprachiasmatic nucleus and intergeniculate leaflet; however, low doses (< 2.0 mg/kg) of S20928 partially blocked the effects of melatonin agonists on most cells. The non-selective serotonin antagonist metergoline did not block the effects of either melatonin agonist. Both melatonin agonists and antagonists were less effective when applied to cells in the hippocampus and dorsal lateral geniculate nucleus. These results indicate that S20098 is an agonist acting probably on melatonin receptors in the Syrian hamster brain. S20928 may have mixed agonist/antagonist properties, but at low doses appears to function as an antagonist at melatonin receptors in the suprachiasmatic nucleus and intergeniculate leaflet.

Acetamides↗
PubMed1994

Effects of serotonergic agonists on firing rates of photically responsive cells in the hamster suprachiasmatic nucleus.

Serotonergic neurons from the midbrain raphe nuclei innervate the suprachiasmatic nucleus (SCN) of the hypothalamus, which functions as the dominant pacemaker for mammalian circadian rhythms. We investigated the effects of serotonin (5-HT) on firing rates of light-activated SCN cells in urethane-anesthetized hamsters. Micro-iontophoretic application of 5-HT or 5-HT1A agonists (8-OH-DPAT and 5-CT) caused a dose-dependent inhibition of spontaneous activity and photic responses in the majority of SCN cells tested. Application of metergoline alone, a non-selective 5-HT antagonist, slightly increased firing rates during darkness and light exposure, suggesting a tonic serotonergic suppression of SCN activity. Metergoline also effectively attenuated suppression induced by the three 5-HT agonists. In addition, the effects of 8-OH-DPAT were blocked by a 5-HT1A antagonist, SDZ 216-525. However, other putative 5-HT antagonists were weak (propranolol and NAN-190) or ineffective (ketanserin) in blocking the action of 8-OH-DPAT. These results indicate that serotonin has a potent role in reducing photic effects on retinally activated SCN cells in hamsters, and that these effects are mediated by a receptor with properties similar to those of the 5-HT1A subtype.

8-Hydroxy-2-(di-n-propylamino)tetralin↗
PubMed1993

Effects of serotonin agonists and melatonin on photic responses of hamster intergeniculate leaflet neurons.

Retinal input to the suprachiasmatic nuclei (SCN) and the intergeniculate leaflet (IGL) is involved in photic entrainment of mammalian circadian rhythms. The activating effects of light on firing rates of IGL cells may be regulated by serotonin (5-HT), since the IGL receives a dense serotonergic input from the midbrain raphe. We investigated the effects of 5-HT agonists and melatonin (a derivative of 5-HT) on single-unit discharges of light-sensitive cells in the hamster IGL area, using a microiontophoretic technique. 5-HT and a 5-HT1A-selective agonist, 8-OH-DPAT, potently suppressed both spontaneous and light-induced activity of IGL cells in a dose-related manner. This suppression was unchanged or potentiated by concurrently applied Mg2+, suggesting a direct action. Furthermore, the suppressive effects of both agonists were antagonized by a nonselective 5-HT antagonist, metergoline, and a 5-HT1A-directed antagonist, pindobind-5-HT1A. However, other putative 5-HT1A antagonists were weak (propranolol) or ineffective (pindolol and spiperone) in blocking the effects of 8-OH-DPAT. Neither of two 5-HT2 antagonists tested was able to block the effects of 5-HT. Melatonin generally mimicked the effects of 5-HT agonists on IGL cells, but these effects were not attenuated by 5-HT antagonists. The results indicate that both 5-HT and melatonin exert inhibitory effects on spontaneous activity and photic responses of cells in the hamster IGL, and that these effects are mediated via a 5-HT1A-like receptor and a melatonin receptor, respectively.

Action Potentials↗
PubMed1993

Human malignant melanoma cells express high-affinity receptors for melatonin: antiproliferative effects of melatonin and 6-chloromelatonin.

In order to explore the potential oncostatic properties of the pineal hormone, melatonin, we have investigated its binding characteristics and functional effects in a human malignant melanoma (M-6) cell line. Binding studies in M-6 membranes showed the coexistence of 2-[125I]iodomelatonin binding sites with picomolar and nanomolar affinities. Guanine nucleotides caused conversion of all high-affinity sites to a low-affinity state without a change in binding capacity. Melatonin induced a marked concentration-dependent reduction in forskolin-stimulated cAMP accumulation in intact M-6 cells, indicating that it binds to a functional receptor in this cell line. The in vitro proliferation of M-6 cells was significantly inhibited by melatonin and its analogues 6-chloromelatonin, and 2-iodomelatonin, at concentrations ranging from 10(-9) to 10(-4) M, as demonstrated by cell counts and measurements of DNA content. These findings indicate that M-6 cells express functional receptors for melatonin which may be involved in mediating the antiproliferative effects of this hormone.

Adenylyl Cyclase Inhibitors↗
PubMed1993

Neurophysiological analysis of circadian rhythm entrainment.

We review recent studies in our laboratory that have investigated the neural mechanisms underlying photic entrainment of the mammalian circadian system. The results from studies of extracellular single-unit recordings and of photic induction of Fos-like immunoreactivity (Fos-lir) indicate that excitatory amino acid (EAA) transmission, and particularly activation of the N-methyl-D-aspartate (NMDA) receptor subtype, is important for conveying photic information to suprachiasmatic nucleus (SCN) cells. We have also found that a subregion of the SCN still shows Fos-lir after blockade of EAA receptors, and we have evidence suggesting that these cells are innervated by a distinct subdivision of the retinal projection to the SCN. In addition, we have found that photic responses of cells in the intergeniculate leaflet (which projects to the SCN) and of SCN cells are modulated by serotonin (5-HT) via a receptor that resembles the 5-HT1A subtype.

Animals↗
PubMed1992

Involvement of multiple sulfhydryl groups in melatonin signal transduction in chick brain.

To gain insight into the molecular mechanism underlying melatonin binding and signal transduction in the chick brain, we have investigated the role of -SH groups, using a sulfhydryl alkylating reagent N-ethylmaleimide (NEM). At least two -SH groups are involved in the formation of the receptor-G protein complex: one is sensitive to and the other relatively insensitive to NEM. Alkylation of the sensitive group selectively abolishes high affinity binding of 2-[125I]iodomelatonin ([125I]MEL), similar to the effect induced by GTP, thus leading to a complete loss of sensitivity to nucleotides. Modification of both groups causes a marked reduction in binding capacity. Agonists with high affinity, but not other compounds with low affinity for the melatonin receptor, protect against alkylation by NEM. GTP gamma s does not significantly alter the reactivity of -SH groups towards NEM, but agonist-protected receptors remain sensitive to this nucleotide. Moreover, NEM pretreatment blocks the inhibitory effect of melatonin on forskolin-stimulated adenylate cyclase activity in chick brain. These data suggest that the -SH group modulating agonist affinity may lie within the coupling domain between the receptor and G protein but outside of the GTP binding site. In addition, sulfhydryl groups are essential for melatonin binding and signal transduction in chick brain.

Adenylyl Cyclases↗
PubMed1991

Pertussis toxin blocks melatonin-induced inhibition of forskolin-stimulated adenylate cyclase activity in the chick brain.

The high-affinity guanine nucleotide-sensitive receptor sites for melatonin in the mammalian hypothalamus and pars tuberalis mediate inhibition of adenylate cyclase (AC) activity. Therefore, we have examined whether similar sites in the chick brain and retina also modulate AC activity. Melatonin did not alter basal or forskolin-stimulated AC activity in whole forebrain or retinal homogenates. In contrast, melatonin significantly inhibited forskolin-stimulated AC activity in forebrain synaptosomal membranes and partially purified retinal membranes in a concentration-dependent manner. Maximal inhibition (approximately 25-30%) of stimulated AC activity was observed at 10-100nM melatonin, while the concentrations (EC50's) which caused half-maximal effects were 22 +/- 6 pM and 30 +/- 5 pM in the brain and retina respectively. Pretreatment of forebrain slices with pertussis toxin abolished the inhibitory effect of melatonin on stimulated AC activity. These data provide the first evidence that melatonin suppresses AC activity in the chick CNS via a pertussis toxin-sensitive G-protein.

Adenylate Cyclase Toxin↗
PubMed1991

3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate-solubilized binding sites for 2-[125I]iodomelatonin in chick brain retain sensitivity to guanine nucleotides.

Binding of 2-[125I]iodomelatonin to 3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate (CHAPS)-solubilized sites from chick forebrain was rapid. reversible, saturable, of high affinity, and of pharmacological selectivity. Scatchard analyses showed that 2-[125I]iodomelatonin binds to a single site with equilibrium dissociation constant (KD) values of 328 +/- 22 (n = 4) and 302 +/- 26 pM (n = 3) and a maximal number of binding sites (Bmax) of 36.2 +/- 2.0 and 49.5 +/- 6.6 fmol/mg of protein in solubilized and membrane fractions, respectively. The KD values obtained from the ratio of kinetic constants (k2/k1) in solubilized and membrane preparations were 228 and 216 pM, respectively. Inhibition studies indicated the following order of pharmacological affinities for both membrane and solubilized sites: 2-iodomelatonin greater than melatonin greater than 6-chloromelatonin much greater than prazosin greater than N-acetylserotonin much greater than serotonin greater than metergoline greater than ketanserin greater than propranolol greater than phentolamine greater than cyproheptadine. Guanyl nucleotides inhibited binding of 2-[125I]iodomelatonin to solubilized and membrane fractions, by converting binding sites from a high-affinity to a low-affinity state. These findings show that solubilized binding sites for melatonin exhibit the specific binding and pharmacological characteristics present in membrane-bound sites. Moreover, the retention of sensitivity to guanine nucleotides in fractions solubilized with CHAPS suggests that this solubilization procedure is suitable for further studies aimed at the isolation, purification, and molecular characterization of active melatonin binding sites.

Animals↗
PubMed1990

[Effects of the pineal body and melatonin on sensitivity to pain in mice].

Sensitivity to painful stimuli was measured by hot plate, writhing tests and electric caudal stimulation in mice. The mice were kept under light-dark 12/12 cycle with light out at 18:00 for at least 2 wk. Both basal analgesia and meperidine (pethidine)-induced analgesic effect exhibited parallel circadian rhythms, with the marked peak and through occurring at mid-dark and mid-light phases, respectively. The day-night differences in pain threshold 10 d after pinealectomy were not evident, especially in the loss of dark time augmentation of analgesic responses, but persisted in sham operated mice. Melatonin (MT) 50-200 mg/kg ip during light phase produced analgesic activity. MT 250 mg/kg ip resulted in a loss of the righting reflex. In pinealectomized mice, the pre-treatment of MT 5 mg/kg potentiated levels of analgesia induced by meperidine 10 mg/kg or morphine 5 mg/kg. It is possible that MT is one of the endogenous nocifensor inhibitors in CNS.

Animals↗