Biomedical subjects

R E Rosenstein

Publications and source records attributed to R E Rosenstein.

At least 19 recordsLinked to original sources

PubMed2006

Melatonin in the eye: implications for glaucoma.

Melatonin synthesis occurs in the retina of most animals as well as in humans. Circadian oscillators that control retinal melatonin synthesis have been identified in the eyes of different animal species. The presence of melatonin receptors is demonstrable by immunocytochemical studies of ocular tissues. These receptors may have different functional roles in different parts of the eye. In view that melatonin is a potent antioxidant molecule, it can be effective in scavenging free radicals that are generated in ocular tissues. By this mechanism melatonin could protect the ocular tissues against disorders like glaucoma, age-related macular degeneration, retinopathy of prematurity, photo-keratitis and cataracts. Although an increased intraocular pressure is an important risk factor in glaucoma, other concomitant phenomena like increased glutamate levels, altered nitric oxide metabolism and increased free radical generation seem to play a significant role in its pathogenesis. Data are discussed indicating that melatonin, being an efficient antioxidant with antinitridergic properties, has a promising role in the treatment and management of glaucoma.

Animals↗

PubMed2001

Circadian activity of the GABAergic system in the golden hamster retina.

Daily changes in gamma-aminobutyric acid (GABA) turnover rate were studied in the golden hamster retina. This parameter showed significant variations throughout the light-dark cycle, with minimal values during the day. Retinal glutamic acid decarboxylase (GAD) activity was higher at midnight than at noon. Moreover, [3H]GABA binding significantly varied throughout the 24-h cycle, with maximal values during the day. Saturation studies performed at 12:00 and 24:00 h indicated that the maximal concentration of [3H]GABA binding sites (Bmax) was significantly higher at noon, whereas the dissociation constant (Kd) remained unchanged. High K+-induced GABA release was significantly higher at midnight than at midday. Daily variations in retinal GABA turnover rate, GABA release, and in its specific binding persisted in golden hamsters exposed to constant darkness. In summary, these results support the idea of a circadian clock-controlled GABAergic activity in the hamster retina.

Animals↗

PubMed2001

Nitrosation of melatonin by nitric oxide: a computational study.

Melatonin is being increasingly promoted as a therapeutic agent for the treatment of jet lag and insomnia, and is an efficient free radical scavenger. We have recently characterized a product for the reaction of melatonin with nitric oxide (NO), N-nitrosomelatonin. In the present work, reaction pathways with N1, C2, C4, C6 and C7 as possible targets for its reaction with NO that yield the respective nitroso derivatives have been investigated using semiempirical AM1 computational tools, both in vacuo and aqueous solution. Specifically, two different pathways were studied: a radical mechanism involving the hydrogen atom abstraction to yield a neutral radical followed by NO addition, and an ionic mechanism involving addition of nitrosonium ion to the indolic moiety. Our results show that the indolic nitrogen is the most probable site for nitrosation by the radical mechanism, whereas different targets are probable considering the ionic pathway. These results are in good agreement with previous experimental findings and provide a coherent picture for the interaction of melatonin with NO.

Animals↗

PubMed2000

Dopamine decreases melatonin content in golden hamster retina.

Dopamine significantly decreased melatonin levels in Golden hamster retinas excised at noon and incubated under light. The effect of dopamine was reversed by spiperone and clozapine (selective antagonists for D(2) and for D(4)/D(2) dopaminergic receptors, respectively) but not by SCH 23390 (a selective D(1) dopamine receptor antagonist). Both clozapine and spiperone per se significantly increased melatonin levels, whereas SCH 23390 was ineffective. Quinpirole (an agonist for D(2)-subfamily dopaminergic receptor) decreased melatonin content in retinas excised at midday. Dopamine increased, whereas quinpirole decreased, cAMP accumulation in retinas excised at noon. Retinal dopaminergic turnover rate (assessed as the ratio of 3,4-dihydroxyphenylacetic acid to dopamine) was significantly higher at midday than at midnight. In retinas excised at midnight, melatonin content in vitro was unaffected by dopamine or quinpirole. At midnight, dopamine increased cAMP accumulation, whereas quinpirole was ineffective. When hamsters were kept under constant darkness for 48 h and sacrificed at subjective midday or midnight, dopamine increased cAMP accumulation at both times, whereas quinpirole decreased this parameter only at subjective midday. Dopaminergic turnover rate was significantly higher at subjective midday than at subjective midnight. These results show that dopamine regulates melatonin biosynthesis in the Golden hamster retina.

3,4-Dihydroxyphenylacetic Acid↗

PubMed2000

Effect of brimonidine on rabbit trabecular meshwork hyaluronidase activity.

PURPOSE: To study the presence of hyaluronidase activity in the rabbit trabecular meshwork and its regulation by brimonidine. METHODS: A spectrophotometric assay that consists of the assessment of N-acetylhexosamine groups released from hyaluronic acid was used to examine hyaluronidase activity. Cyclic adenosine monophosphate (cAMP) levels were assessed by radioimmunoassay. RESULTS: Hyaluronidase activity was detected in the rabbit trabecular meshwork. Its optimal activity was in the acid range of pH 3.8. Brimonidine significantly increased trabecular hyaluronidase-specific activity and decreased cAMP accumulation. Yohimbine significantly inhibited the effect of brimonidine on both hyaluronidase activity and cAMP accumulation. CONCLUSIONS: The finding of endogenous hyaluronidase activity in rabbit trabecular meshwork supports the hypothesis that this tissue can metabolize its own glycosaminoglycan (GAG) products. The present results suggest, however, that the hypotensive effect of brimonidine could be mediated, at least in part, by its ability to increase GAG catabolism, probably through a cAMP-independent mechanism.

Adrenergic alpha-Agonists↗

PubMed1999

Effect of GABA on melatonin content in golden hamster retina.

The effect of GABA on melatonin content in vitro was studied in the golden hamster retina. GABA significantly increased melatonin levels in a dose-dependent manner, its effect being reversed by a GABA(A) receptor antagonist, bicuculline, but not by saclofen, a GABA(B) antagonist. Moreover, an equimolar concentration of muscimol, a GABA(A) receptor agonist, significantly increased retinal melatonin content, whereas baclofen, a GABA(B) receptor agonist, was ineffective. The darkness-induced increase in melatonin content in vitro was inhibited by bicuculline, whereas saclofen was ineffective. Retinal GABA turnover rate was significantly higher at midnight than at midday. GABA significantly decreased cyclic AMP and increased cyclic GMP accumulation in the golden hamster retina. The effect of GABA on both nucleotide levels was reversed by bicuculline, but baclofen had no effect. Cyclic GMP analogues (i.e., 8-bromoguanosine 3',5'-cyclic monophosphate and 2'-O-dibutyrylguanosine 3',5'-cyclic monophosphate) significantly increased retinal melatonin content in vitro. Taken together, these results support the hypothesis that GABA may be important for the "dark message" in the hamster retina.

1-Methyl-3-isobutylxanthine↗

PubMed1999

GABA release mechanism in the golden hamster retina.

High K+ medium and glutamate elicited a significant [3H]-GABA release in the golden hamster retina. High K+ -induced GABA release was largely calcium-dependent, while the effect of glutamate was Ca2+ -independent. After replacing Na+ by Li+, glutamate-evoked [3H]-GABA release was abolished, while high K+ -evoked release remained unchanged. The effect of glutamate was completely blocked by DNQX but not by APV. Furthermore, kainate induced [3H]-GABA release, whereas NMDA was ineffective. Assessment of endogenous GABA efflux further confirmed results obtained for [3H]-GABA. GABA-like immunoreactivity was observed in amacrine cells, in neurons localized in ganglion cell layer, as well as in fibers and terminals at the inner plexiform layer. In addition a few horizontal cells showed GABA-like immunolabeling. The present results suggest the existence of at least two pools of GABA in the hamster retina, compatible with both vesicular and carrier-mediated mechanisms of transmitter release, being the amacrine cells the main gabaergic source in this tissue.

Animals↗

PubMed1998

Reactions of melatonin and related indoles with free radicals: a computational study.

Melatonin is being increasingly promoted as a therapeutic agent for the treatment of jet lag and insomnia and has been recently suggested to act as an efficient free-radical scavenger. In the present work, its mechanisms of action for scavenging hydroxyl radicals have been investigated using semiempirical AM1 and density functional theory (DFT) computational tools. Two different reactions were proposed as follows: one involving the abstraction of an indolic hydrogen to yield a neutral radical and another involving the addition of the hydroxyl radical to the indolic moiety. Our results show that, from a thermodynamical standpoint, melatonin may directly scavenge hydroxyl radicals both in vacuum and in aqueous solution. The structural requirements for free-radical-trapping ability have been examined comparing melatonin with related indoles. Computational data suggest that 5-methoxy and N-acetyl groups of melatonin do not significantly affect its thermodynamical capacity of free-radical trapping. The present results support experimental data on the potential of melatonin as a physiological or pharmacological antioxidant agent.

Computer Simulation↗

PubMed1998

Distribution of GABA in the golden hamster retina. An immunocytochemical study.

Distribution of the inhibitory neurotransmitter GABA was studied in the golden hamster retina using immunocytochemistry at cellular and subcellular levels. GABA-immunoreactivity was observed in somata of amacrine, displaced amacrine and horizontal cells. GABA immunoreactive fibers were abundant in the inner plexiform layer. Ultrastructural analysis exhibited dense GABA-immunoreactive deposits in amacrine cell somata, processes and terminals. Immunolabelling was also observed in the cytoplasm of horizontal or interplexiform cells and displaced amacrine cells. In every case DAB deposits were observed in the cytosolic compartment, attached to the inner surface of cell membranes and to outer mitochondrial membranes. Immunolabeled terminals predominated in the inner plexiform layer and immunoprecipitates were also observed attached to the outer face of vesicle membranes as well as completely filling synaptic vesicles. Both clear and dense core vesicles were observed. The present results are similar to those obtained in other mammalian species showing GABA immunoreactivity in amacrine, displaced amacrine and horizontal cells.

Animals↗

PubMed1997

Photic control of nitric oxide synthase activity in golden hamster retina.

A characterization of nitric oxide synthase (NOS) in the golden hamster retina was performed. Enzymatic activity was partially Ca2+ and calmodulin-dependent, required NADPH, and was inhibited by L-arginine analogs. Retinal NOS activity was higher at midnight than at midday. When the hamster were placed under constant darkness for 24 or 48 h, and killed at equivalent time points, representing subjective day and subjective night respectively, the differences in NOS activity disappeared. One hour of darkness during the day increased, while the same period of light during the night decreased, retinal NOS activity. From these results, it might be presumed that hamster retinal NOS activity is regulated by the photic stimulus.

Animals↗

PubMed1997

Aging attenuates diurnal variation in hamster locomotion, anxiolysis and GABA turnover.

A decrease in amplitude of wheel running circadian rhythms was found in old (18 month old) Syrian hamsters, as compared with young (3 month old.) animals. In a plus-maze paradigm, amplitude of variation of anxiety-related variables (2400 vs. 1600 h) was significantly impaired in aged hamsters. Cerebral cortex, hypothalamic and pineal gamma-aminobutyric acid (GABA) turnover was higher at night, amplitude of variation being significantly smaller in aged hamsters. The results further support the existence of impaired amplitude of circadian rhythms in aged Syrian hamsters.

Age Factors↗

PubMed1997

Melatonin site and mechanism of action: single or multiple?

By affecting the entrainment pathways of the biologic clock, melatonin has a major influence on the circadian and seasonal organization of vertebrates. In addition, a number of versatile functions that far transcend melatonin actions on photoperiodic time measurement and circadian entrainment have emerged. Melatonin is a free radical scavenger and antioxidant and it has a significant immunomodulatory activity, being presumably a major factor in an organism's defense toxic agents and invading organisms. Besides affecting specific receptors in cell membranes to exert its effects, the interaction of melatonin with nuclear receptor sites and with intracellular proteins, like calmodulin or tubulin-associated proteins, as well as the direct antioxidant effects of melatonin, may explain many general functions of the pineal hormone.

Animals↗

PubMed1996

Melatonin effect on the cyclic GMP system in the golden hamster retina.

Melatonin effect on retinal cyclic GMP accumulation, guanylate cyclase activity, cyclic GMP content and cyclic GMP phospho-diesterase activity was examined in the Syrian hamster retina. Melatonin increased significantly cyclic GMP accumulation at picomolar concentrations and in a time-dependent manner. The kinetic analysis of guanylate cyclase activity revealed a significant increase of both apparent Vmax and K(m), induced by 10 nM melatonin. The effect of melatonin was higher in the absence, than in the presence of the phoshodiesterase inhibitor (IBMX), suggesting an effect on cyclic GMP catabolism. Phosphodiesterase activity was significantly decreased by melatonin. The results show a dual effect of melatonin on cyclic GMP levels, i.e. by increasing the synthesis and inhibiting the degradation, both resulting in an increase of cyclic GMP levels. Taking into account the key role of cyclic GMP in visual mechanisms, the results would suggest the participation of melatonin in retinal physiology.

Animals↗

PubMed1996

Daily variations in cGMP, guanylate cyclase and phosphodiesterase in the golden hamster retina.

Daily variations in cGMP, guanylate cyclase and phosphodiesterase activity in golden hamster retina were studied. Cyclic GMP content exhibited significant variations throughout the 24-hr cycle with maximal values during the dark phase. In order to establish the relative participation of nucleotide synthesis and breakdown during a 24-hr cycle, guanylate cyclase and phosphodiesterase activity were measured in hamsters killed at eight intervals. Guanylate cyclase activity increased at night, peaking at 22.00 hr. Phosphodiesterase activity did not change significantly throughout the light-dark cycle. Light exposure during the night inhibited the nocturnal increase in cGMP content and guanylate cyclase activity, while phosphodiesterase remained unchanged. From these results, it might be presumed that in response to continuous (in a range of hr) light or dark stimuli, the retina would process the photic signal in a different way from that in the short term (in a range of msec).

Animals↗

PubMed1996

Melatonin effect on [3H] glutamate uptake and release in the golden hamster retina.

The effect of melatonin on [3H]glutamate uptake and release in the golden hamster retina was studied. In retinas excised in the middle of the dark phase, i.e., at 2400 h, melatonin (0.1 and 10 nM) significantly increased [3H]glutamate uptake, and this effect persisted in a Ca2+-free medium. On the other hand, melatonin significantly increased [3H]glutamate release in retinas excised at 2400 h, but this effect was Ca2+ sensitive. Melatonin significantly increased 45Ca2+ uptake by a crude synaptosomal fraction from retinas of hamsters killed at 2400 h. In retinas excised at 1200 h, melatonin had no effect on [3H]glutamate uptake, [3H]glutamate release, or 45Ca2+ uptake at any concentration tested. Cyclic GMP analogues, i.e., 8-bromoguanosine 3',5'-cyclic monophosphate and 2'-O-dibutyrylguanosine 3',5'-cyclic monophosphate, significantly increased [3H]-glutamate uptake, [3H]glutamate release, and 45Ca2+ uptake by tissue removed at 1200 and 2400 h, suggesting that the effects of melatonin could correlate with a previously described effect of melatonin on cyclic GMP levels in the golden hamster retina. Taking into account the key role of glutamate in visual mechanisms, the results suggest the participation of melatonin in retinal physiology.

Animals↗

PubMed1995

Daily variations in 2-[125I]melatonin specific binding in the golden hamster retina.

Daily variations in melatonin content and 2-[125I]melatonin specific binding in retinas of golden hamsters were studied. Both parameters showed significant variations throughout the 24 h period. Maximal specific binding was observed at 24.00 h, while melatonin content peaked at 04.00 h. Saturation studies performed at 12.00 and 24.00 h indicated that the maximal concentration of 2-[125I]melatonin binding sites (Bmax) was significantly higher at 24.00 h than at 12.00 h, whereas the dissociation constant (Kd) remained unchanged. As 2-[125I]melatonin specific binding decreased at times when retinal melatonin content was high, these findings suggest that daily variations in retinal melatonin levels may be implicated in the regulation of the density of melatonin binding sites, probably through mechanisms of up- and down-regulation induced by melatonin on its own binding sites.

Analysis of Variance↗

PubMed1995

Daily variations in GABA receptor function in Syrian hamster cerebral cortex.

The existence of diurnal changes in postsynaptic expression of gamma-aminobutyric acid (GABA) type A receptors was assessed in cerebral cortex of Syrian hamsters by measuring [3H]GABA binding and the influx of 36Cl- in synaptoneurosomes. A diurnal variation in dissociation constant of [3H]GABA binding to cerebral cortex membranes, and the absence of diurnal differences in maximal number of sites, were found. When the nycthemeral changes in muscimol-stimulated 36Cl- uptake by cortical synaptoneurosomes were assessed, a maximum occurred late at night (i.e. 0400 h). At 1600 h, micromolar concentrations of flunitrazepam potentiated significantly the influx of chloride induced by muscimol, while at 0400 h flunitrazepam did not exert any significant effect on 36Cl- uptake. The results indicate that postsynaptic type A GABAergic activity peaked at nocturnal hours in the cerebral cortex of Syrian hamsters.

Animals↗

PubMed1995

Evidence for local synthesis of melatonin in golden hamster retina.

Daily variations in melatonin content of retinas of pinealectomized and sham-operated golden hamsters were studied. Melatonin content showed significant daily variations with maximal values at night (i.e. early in the night in pinealectomized hamsters and late at night in sham-operated animals). Moreover, mean retinal melatonin levels augmented significantly after pinealectomy. In vitro the augmented melatonin levels found in retinas incubated in darkness for 8 h was suppressed by exposure to light, indicating the ability of hamster retina to regulate melatonin synthesis in isolated conditions. Taken together, the in vivo and in vitro results support daily variations of melatonin content of exclusive retinal origin.

Analysis of Variance↗