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Biomedical subjects

M Urbina

Publications and source records attributed to M Urbina.

At least 37 records · Page 2Linked to original sources

Synthesis of serotonin from 5-hydroxytryptophan in the post-crush retina: inhibition of in vitro outgrowth by the intraocular administration of the precursor.

Serotonin is present in the retina of many species, in which plays roles as a neurotransmitter, as a modulator of regeneration, and as the precursor of melatonin. The turnover of serotonin in the goldfish retina is modified by the lesion of the optic nerve and, in postcrush goldfish retinal explants, serotonin inhibits the outgrowth. In the present study, the modification of the serotonergic system of the retina induced by the process of regeneration was explored. The addition of the precursor of serotonin, 5-hydroxytryptophan, to retinal explants, increased the levels of serotonin in a concentration-dependent manner. The concentration of serotonin differentially increased in control and postcrush explants cultured in the presence of 5-hydroxytryptophan for various periods of time, indicating a greater accumulation of the indoleamine at early periods of time in the control than in the postcrush tissue culture. This observation, together with the fact that serotonin concentration in postcrush retina cultured in the absence of 5-hydroxytryptophan and exposed to the precursor for 60 min increased less than in the control, indicates a saturation of the serotonergic system produced by the lesion. The addition of imipramine or citalopram, serotonin uptake blockers, did not significantly change the concentration of serotonin in the cultures, thus, the elevation of serotonin accumulation, especially in the post-crush tissue, might not be due to the transport from the medium. The intraocular injection of 5-hydroxytryptophan after the crush of the optic nerve resulted in a decrease in the outgrowth of retinal explants, supporting the in vivo role of serotonin during the regenerating process in situ. The lesion of the optic nerve did not affect the specific cells, since the number of serotonin-immunoreactive neurons in the retina were not modified by the crush. Taken together, retinal serotonin system is regulated after producing a lesion of the optic nerve, a modulation which has been demonstrated in vivo and in vitro. Thus, there is a reciprocal interaction, since serotonin influences outgrowth in the postcrush retina and the serotonergic system is modulated by the crush, indicating a mechanism of feed-back regulation.

5-Hydroxytryptophan↗

5HT1A receptor agonist differentially increases cyclic AMP concentration in intact and lesioned goldfish retina. In vitro inhibition of outgrowth by forskolin.

5HT1A receptors occur in the retina of various species and the administration of 5HT1A agonists results in the inhibition of outgrowth from postcrush goldfish retinal explants. The levels of cyclic AMP (cAMP) play a role in the modulation of the outgrowth of the nevous system. Moreover, the stimulation of central 5HT1A receptors with the agonist 8-hydroxy-2-(di-n-propylamino)tetralin has been reported to produce an increase or decrease in the activity of adenylate cyclase. In the present investigation we studied the effect of adenylate cyclase stimulation by forskolin, as well as the modulatory effects of 5HT1A receptor agonists and antagonists on the production of cAMP in the goldfish retina, and on the outgrowth of this tissue in vitro. 8-Hydroxy-2-(di-n-propylamino)tetralin produced a significant and dose-dependent increase in cAMP concentration. This effect was not additive to the stimulation produced by forskolin. By contrast, as previously described, the 5HT1A agonist decreased cAMP concentration in the hippocampus of the rat. Both effects were significantly impaired by the 5HT1A antagonist WAY-100,135. A significant effect of the antagonist alone was observed only in the goldfish retina. The increase in cAMP levels was greater in the intact than in the postcrush retina. In addition, forskolin decreased the outgrowth of postcrush retinal explants in a dose-dependent manner, suggesting the importance of critical levels of cAMP in this process. Taken together, 5HT1A receptors seem to be positively coupled to adenylate cyclase in the goldfish retina, where cAMP plays a role as a modulator of outgrowth and regeneration. The inhibitory effect of 5HT1A receptor agonists on retinal outgrowth might be mediated through the production of cAMP. The activation of other subtypes of 5HT receptors positively coupled to adenylate cyclase by the 5HT1A agonist, such as 5HT7, cannot be discarded.

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

Serotonin turnover rate, [3H]paroxetine binding sites, and 5-HT1A receptors in the hippocampus of rats subchronically treated with clonazepam.

Selective central benzodiazepine agonists, such as clonazepam, are known to modify serotonin and 5-hydroxyindoleacetic content in the brain. In order to further study the effect of this benzodiazepine on serotonin turnover rate, rats received clonazepam, 10 mg/kg for 10 days, and the concentrations of serotonin and 5-hydroxyindoleacetic acid were determined in the hippocampus after inhibition of monoamineoxidase with pargyline. The results indicate a reduction in the turnover rate of the monoamine. In addition, the systemic administration of clonazepam produced a decrease in the Bmax of [3H]DPAT binding to 5-HT1A sites in the hippocampus. By contrast, this effect was not observed if clonazepam was delivered into the dorsal raphe nucleus by osmotic minipumps. The binding of [3H]paroxetine to 5-HT reuptake sites was increased by the treatment with clonazepam. The present observations indicate that clonazepam produces a reduction of serotonin turnover rate in the hippocampus of the rat concomitant with a down-regulation of 5-HT1A binding sites, probably by an effect at the forebrain projections. There is also an up-regulation of the serotonin transporter, which might contribute to a reduction in the synaptic availability of serotonin during clonazepam treatment.

Animals↗

Increase of the serotonin metabolite, 5-hydroxyindoleacetic acid, in cerebro-spinal fluid and spinal cord of bovines affected with the paraplegic syndrome.

The paraplegic syndrome of bovines is a condition characterized by impairment of locomotion, hypoalgesia and finally death within 72 h. The pathogenesis of the syndrome has not been established. In the present work we determined the levels of monoamines and their metabolites in cerebro-spinal fluid and spinal cord of affected animals in order to investigate the functional state of these neurotransmitters. The content of the main metabolite of serotonin, 5-hydroxyindoleacetic acid, was elevated in the cerebro-spinal fluid and in the gray matter of the spinal cord of paraplegic bovines. Serotonin content in the spinal cord did not differ with respect to control animals, but was decreased in the cerebro-spinal fluid of affected animals. Modifications in the noradrenergic system were also observed, but were less consistent, for which reason further studies are needed. These observations indicate an increase in the turnover rate of serotonin in the paraplegic syndrome. The meaning of the described alterations is unknown at the moment.

Animals↗

Dopamine and serotonin turnover rate in the retina of rabbit, rat, goldfish, and Eugerres plumieri: light effects in goldfish and rat.

The concentration of dopamine, and its metabolites 3,4-dihydroxyphenylacetic and homovanillic acids, as well as serotonin and its metabolite 5-hydroxyindoleacetic acid, were determined in the retina of two teleosts, C. auratus (goldfish) and E. plumieri (mojarra), and two mammals, R. norvegicus (rat) and O. cuniculus (rabbit). The turnover rate of these monoamines were investigated in the four species by the calculation of the ratio monoamine/metabolite as an indirect index, and in goldfish and rat by the inhibition of the synthesis with alpha-methyl-p-tyrosine or p-chlorophenylalanine, by the increase in dopamine or serotonin by the corresponding precursors, 3,4-dihydroxyphenylalanine or 5-hydroxytryptophan, and by inhibition of monoaminooxidase with pargyline. The modulation by light and dark stimulation was studied in the goldfish and the rat. Differences in the concentration and turnover rate were observed among the species. Serotonin concentration was higher in the teleosts. The administration of inhibitors of dopamine and serotonin synthesis differentially decreased the levels of the monoamines in the retina of goldfish and rat. The rate of formation of dopamine and serotonin by the corresponding precursors was much higher in the goldfish than in the rat. Pargyline administration decreased 3,4-dihydroxyphenylacetic and 5-hydroxyindoleacetic acids at different rates and time dependency in the retina of goldfish and rat. Dopamine and serotonin concentration did not exhibit high modifications by the inhibitor, suggesting the function of regulatory mechanisms or additional effect of pargyline at other sites different from monoaminooxidase.(ABSTRACT TRUNCATED AT 250 WORDS)

3,4-Dihydroxyphenylacetic Acid↗

Serotonin inhibits outgrowth of goldfish retina and impairs the trophic effect of taurine.

The regeneration of explants prepared from goldfish retinas with a prior crush of the optic nerve is stimulated by the sulphur amino acid, taurine. Serotonin has been reported to modify survival, proliferation, and outgrowth of nervous tissue. In the present work we evaluated the effect of serotonin and some serotonergic agonists on the neuritic outgrowth from goldfish retinal explants. Serotonin, its precursor, 5-hydroxytryptophan, and the 5HT1A receptor agonists, 8-hydroxy-2-(di-n-propylamino)tetralin and buspirone, inhibited the outgrowth. The blockers of serotonin uptake, imipramine and citalopram, were also inhibitors of neurite sprouting. Imipramine favoured the inhibitory effect of serotonin at 10 days in culture. The concentration of serotonin and its metabolite, 5-hydroxyindoleacetic acid, decreased in the retina at 3 and 5 days after the crush of the optic nerve. Serotonin levels started to recover after 5 days post-lesion, and the metabolite also increased. This indicates that the lesion increases the turnover rate of serotonin and this may be related to its role in regeneration. Serotonin concentration was elevated by the intraocular administration of its precursor, 5-hydroxytryptophan, indicating that the capacity for synthesis was preserved after the crush, but that it was smaller in the post-lesioned retinas. The trophic effect of taurine was impaired by a low concentration of serotonin, probably by opposing the final effect on growth via different targets. These results support a role of serotonin in the regeneration of goldfish retina probably through 5HT1A receptors.

5-Hydroxytryptophan↗

8-[3H]hydroxy-2-(di-n-propylamino) tetralin binding sites in goldfish retina.

The binding sites of 8-[3H]hydroxy-2-(di-n-propylamino)tetralin ([3H]DPAT) were characterized in the retina of goldfish in order to evaluate the selectivity of the ligand for serotonin1A (5HT1A) receptors. Specificity of the binding was performed in the presence of serotonergic and dopaminergic agonists and antagonists. Buspirone, spiroxatrine and 5-methoxy-N,N-dimethyltryptamine were potent inhibitors, followed by propranolol, citalopram, imipramine and desipramine. Serotonin was not a potent inhibitor, and its interaction with the binding sites of [3H]DPAT was complex. Nomifensine displayed an important inhibition, however, other dopamine uptake blockers, such as bupropion and GBR-12909, were less potent. Haloperidol was also a good inhibitor, but the D1 receptor agonist, SKF-38393, the D2 receptor antagonist, sulpiride, and dopamine did not inhibit the binding. GppNHp inhibited the binding in the micromolar range. The analysis of saturation experiments by isotopic dilution, using buspirone to determine nonspecific binding, revealed two sites. The number of binding sites defined by buspirone were higher than the ones defined by nomifensine. The specific binding, using buspirone for definition, was reduced by the intraocular injection of 6-hydroxydopamine. This investigation demonstrates that [3H]DPAT labels 5HT1A receptors in goldfish retina, but also interacts with a non-5HT receptor site. These receptors seem to be localized in dopaminergic neurons.

Animals↗

Asymptomatic congenital syphilis and auditory brainstem response.

Untreated congenital syphilis is a known cause of hearing loss. In order to ascertain if infants are at risk of hearing loss at birth, 75 neonates with serologic evidence of syphilis and APGAR scores greater than 9 at 5 min were examined by auditory brainstem response (ABR) audiometry. All of the infants had normal ABRs. Ophthalmologic examination revealed no evidence of interstitial keratitis. Lumbar puncture was attempted on all the neonates. Cerebrospinal fluid (CSF) was successfully obtained from 41 infants and revealed no abnormalities. This study demonstrates that asymptomatic infants with serologic evidence of syphilis do not require audiologic evaluation in the neonatal period.

Audiometry↗

[Detection of Borrelia burgdorferi antibodies in a population sample of the state of Zulia].

Between July 1992 and September 1993 an investigation was carried out in a population of Zulia State, Venezuela, in order to detect antibodies against Borrelia burgdorferi for the diagnosis of Lyme disease. A total of 74 patients were studied: 37 asymptomatic and 37 patients clinically suspected having the disease. ELISA tests were performed to determine antibodies against Borrelia burgdorferi. The positive cases, confirmed by duplicate, were tested with VDRL, Monotest, Rheumatoid Factor and Antinuclear Antibodies to eliminate false positives. The total positive cases were 14 of 74 (18.9%). Positive cases in the symptomatic group (29.7%) were higher than in the asymptomatic group (8.9%). The most frequent clinical diagnosis was Morfea (54.5%). The major serological diagnosis (54.32%) was obtained from the chronic patients (more than a year of evolution). A 45.5% of symptomatic patients presented antibodies, despite of receiving antibiotic treatment. Most of the symptomatic positive cases, and also the asymptomatic cases, had a previous visit or permanence in forestal or rural areas. The results of this investigation prove the existence of antibodies against Borrelia burgdorferi in the population of Zulia State, both in the symptomatic as well as in the asymptomatic patients. These results open the path to use a more specific test like immunoblot, for the diagnosis of Lyme disease in our area.

Adolescent↗

Possible neurotransmitter role of noradrenaline in the teleost retina.

The role of dopamine as a neurotransmitter in the retina of different species has been clearly established; however, there is still some controversy as to whether noradrenaline (NA) is present as a neurotransmitter in this tissue. In this study, we show that, under controlled conditions, NA is present in the retina of goldfish at a concentration of 0.15 +/- 0.03 ng/mg protein and its biosynthetic enzyme, dopamine beta-hydroxylase shows an activity of 2.5 +/- 0.2 pmol NA/hr/mg protein. The amount of NA increases to 1.88 +/- 0.24 ng/mg protein in light adapted animals and decreases to undetectable levels in dark adapted ones. By contrast, dopamine-beta-hydroxylase levels are not affected by changes in light conditions. This finding provides further evidence in favor of a neurotransmitter role for NA in vertebrate retina.

Adaptation, Physiological↗

Retinal neurotransmitter interaction as reflected in horizontal cell spatial behaviour.

The effects of 5-hydroxytryptamine (5-HT) and its precursors 5-hydroxytryptophan (5-HTP) and L-tryptophan (L-Tryp) on the spatial properties of horizontal cells were studied in the isolated and perfused retina of the teleost Eugerres plumieri. All three compounds produce a contraction of the receptive field, evaluated by the ratio of responses evoked by local and distant light stimuli. This is the result of cell uncoupling, revealed by the hindrance to diffusion of intracellularly injected Lucifer yellow. Similar effects are produced by dopamine (DA) and the effectiveness is DA much greater than 5-HT greater than 5-HTP greater than L-Tryp. All these effects are blocked by Haloperidol. HPLC studies of endogenous DA release reveal that it occurs when isolated retinas are incubated with 50 mM potassium, 10 microM 5-HT or 5-HTP, but is not found with up to 1 mM L-Tryp. The results indicate that indolaminergic cells induce the release of DA from interplexiform cells, which in turn uncouples horizontal cells in the fish retina.

5-Hydroxytryptophan↗

Dopamine and noradrenaline content in fish retina: modulation by serotonin.

The presence of noradrenaline (NA) and the possible interaction of serotonin (5-HT) with dopaminergic and noradrenergic neuronal elements was studied in the retina of the teleost Eugerres plumieri. By means of a histofluorescent technique, paired amacrine cells can be visualized after intravitreal injection of NA or 5,6-dihydroxytriptamine, suggesting their probable catecholaminergic and indoleaminergic nature. With a high-performance liquid chromatographic (HPLC) method and after pargyline treatment of the animal, 6.86 ng/mg protein of dopamine (DA) was detected, while NA content was 0.50 ng/mg protein. The NA levels of the retina increased, whereas the DA levels decreased in a significant manner after in vivo treatment with 5-HT. 5-methoxy-N,N-dimethyltryptamine, a 5-HT agonist, was able to mimic this effect partially, while the agonists tryptamine and quipazine did not affect the levels of DA and NA. The antagonists methysergide, metergoline, and cyproheptadine significantly blocked the 5-HT-induced NA increase, whereas only the first two antagonists were effective in inhibiting the 5-HT-induced DA decrease. The 5-HT modulation of NA and DA is apparently receptor mediated and is not due to a hetero-exchange, since imipramine was not able to block the 5-HT effect. These findings support the suggested interaction between serotoninergic and dopaminergic cells [Negishi et al: J Neurosci Res 5:621-635, 1980; Neurosci Lett 25:1-5, 1981]. Furthermore, they demonstrate the possible modulation which 5-HT exerts on the endogenous levels of NA.

Animals↗

Serotonin containing neurons in the retina of the teleost Eugerres plumieri.

The concentration and localization of serotonin was determined in the retina of the teleost Eugerres plumieri by using high performance liquid chromatography (HPLC) and immunohistochemical techniques. Serotonin and dopamine were measured simultaneously, their concentrations in the retina being 77 +/- 8 and 516 +/- 23 ng/mg tissue respectively. Treatment of the animals with pargyline significantly increased the levels of dopamine and serotonin. When retinas were treated with the neurotoxin 5,6-dihydroxytryptamine, the level of serotonin was reduced by more than 90% while the dopamine content only diminished by 20% when compared to controls. By using immunohistochemistry with a monoclonal anti-serotonin antibody it was possible to localize this amine in cell bodies of a population of amacrine cells with processes extending mainly into a thin layer of the most external lamina of the inner plexiform layer. Very few ramifications were seen projecting to the internal lamina of this layer. When visualized in flat mount preparations, dense arborization of fluorescent processes was observed. This is the first direct evidence that serotonin is apparently present in amacrine cells of the retina of E. plumieri with a distribution of the serotonergic terminals similar to goldfish but somewhat different when compared to other species.

5,6-Dihydroxytryptamine↗

Fate of Bacillus sphaericus 1593 and 2362 spores used as larvicides in the aquatic environment.

Dry powders produced from insecticidal Bacillus sphaericus strains 1593 and 2362 were applied against Culex tarsalis and Anopheles franciscanus mosquito larvae in small-plot field trials. Good control of C. tarsalis, but not of A. franciscanus, was produced at 0.1 and 0.2 lb/acre [ca. 0.122 and 0.244 kg/ha]. B. sphaericus spores settled rapidly from upper water layers and accumulated in bottom muds. Control of third/fourth-instar C. tarsalis larvae was maintained through day 4 after testing and was related to the presence of at least 100 spores per ml in the upper water layer 2 days earlier. B. sphaericus was shown to recycle in dead larvae both in the laboratory and in the field, producing an increase of 100- to 1,000-fold in spore numbers. There was no evidence of recycling in treated water nor of significant spore persistence upon reflooding of ponds after a very hot, dry period.

Animals↗

Regional changes in phenylethanolamine-N-methyltransferase of rat brain during development.

The changes of phenylethanolamine-N-methyltransferase (PNMT, EC 2.1.1.28), the enzyme that catalyzes the final step in the biosynthesis of adrenaline, were studied during the development of several regions of rat brain. PNMT is present in medulla oblongata-pons, hypothalamus, cerebellum, and midbrain five days before birth, and a progressive increase in the enzyme activity is observed during development. The adult levels are attained between 15 and 20 days, depending on the region. The increases in PNMT activity in the rostral regions are higher than in the caudal regions. PNMT attains adult levels earlier than tyrosine hydroxylase and dopamine-beta-hydroxylase. The apparatus for adrenaline synthesis seems to be mature at three weeks after birth in the medulla oblongata-pons, which contains the cell bodies of adrenaline-containing neurons. In the other regions the adult levels of enzyme activity are attained at 15 days after birth.

Aging↗

Some properties of phenylethanolamine-N-methyltransferase of rat brain.

Phenylethanolamine-N-methyltransferase (PNMT, EC 2.1.1.28) was partially purified from rat brain. Brain homogenates were subjected to ultracentrifugation, salt fractionation, and gel filtration on Sephadex G-100. To compare the rat brain PNMT with that of adrenals, the same procedure was carried out with rat adrenal homogenates. The brain enzyme was eluted from Sephadex as a single fraction with a molecular weight of 26,900, while the enzyme from adrenals under the same conditions appeared in two fractions with molecular weights of 38,700 and 108,500. The brain fraction separated on Sephadex G-100 was active on phenylethanolamine substrates and inactive on indoleamine and phenylethylamine substrates. Products of the enzyme reaction were identified by bidimensional thin-layer chromatography as N-methyl derivatives of the corresponding amines. Kinetic studies showed that the type of inhibition of PNMT from rat brain and rat adrenals by SK&F 7698 was the same as described for PNMT from rabbit adrenals. Also, when normetanephrine and S-adenosyl-L-methionine were used as substrates, the apparent Km values found with PNMT from rat adrenals and rat brain were similar.

Adrenal Glands↗