Angel Alonso-gómez - Academia.edu (original) (raw)

Papers by Angel Alonso-gómez

Netherlands Journal of Zoology, 1994

ABSTRACT The day/night changes in thyroxine (T4) and triiodothyronine (T3) whole-body concentrati... more ABSTRACT The day/night changes in thyroxine (T4) and triiodothyronine (T3) whole-body concentrations in Rana perezi and Xenopus laevis throughout ontogeny have been studied. Daily cycles in thyroid hormones (TH) with higher nocturnal values appear during premetamorphosis in both species. Cyclicity disappears for T3, while it is reversed for T4, in prometamorphic larvae. In X. laevis higher nocturnal T3 and T4 values were observed at the end of climax.

PLOS ONE, 2015

Ghrelin is a gut-brain peptide hormone, which binds to the growth hormone secretagogue receptor (... more Ghrelin is a gut-brain peptide hormone, which binds to the growth hormone secretagogue receptor (GHS-R) to regulate a wide variety of biological processes in fish. Despite these prominent physiological roles, no studies have reported the anatomical distribution of preproghrelin transcripts using in situ hybridization in a non-mammalian vertebrate, and its mapping within the different encephalic areas remains unknown. Similarly, no information is available on the possible 24-h variations in the expression of preproghrelin and its receptor in any vertebrate species. The first aim of this study was to investigate the anatomical distribution of ghrelin and GHS-R1a ghrelin receptor subtype in brain and gastrointestinal tract of goldfish (Carassius auratus) using immunohistochemistry and in situ hybridization. Our second aim was to characterize possible daily variations of preproghrelin and ghs-r1 mRNA expression in central and peripheral tissues using real-time reverse transcription-quantitative PCR. Results show ghrelin expression and immunoreactivity in the gastrointestinal tract, with the most abundant signal observed in the mucosal epithelium. These are in agreement with previous findings on mucosal cells as the primary synthesizing site of ghrelin in goldfish. Ghrelin receptor was observed mainly in the hypothalamus with low expression in telencephalon, pineal and cerebellum, and in the same gastrointestinal areas as ghrelin. Daily rhythms in mRNA expression were found for preproghrelin and ghs-r1 in hypothalamus and pituitary with the acrophase occurring at nighttime. Preproghrelin, but not ghs-r1a, displayed a similar daily expression rhythm in the gastrointestinal tract with an amplitude 3-fold higher than the rest of tissues. Together, these results described for the first time in fish the mapping of preproghrelin and ghrelin receptor ghs-r1a in brain and gastrointestinal tract of goldfish, and provide the first evidence for a daily regulation of both genes expression in such locations, suggesting a possible connection between the ghrelinergic and circadian systems in teleosts.

Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology, Jan 3, 2015

Glucocorticoids have been recently proposed as input signals of circadian system, although the un... more Glucocorticoids have been recently proposed as input signals of circadian system, although the underlying molecular mechanism remains unclear. This work investigates the role of glucocorticoids as modulators of clock genes expression in the liver of goldfish. In fish maintained under a 12L:12D photoperiod, an intraperitoneal injection at Zeitgeber Time 2 of a glucocorticoid analog, dexamethasone (1 μg/g body weight) induced per1 genes while decreased gbmal1a and gclock1a expression in the liver at 8 h post-injection. A 4-h in vitro exposure of goldfish liver to cortisol (0.1-10 μM) also induced gper1 genes in a concentration-dependent manner. Similarly, the exposure of the goldfish cultured liver to dexamethasone produced a concentration-dependent induction of gper1 genes. Moreover, this glucocorticoid analog led to a decrease in gbmal1a and gclock1a transcripts, while the other clock genes analyzed were unaffected. The induction of gper1a and gper1b by dexamethasone in vitro was ob...

Journal of Neurochemistry

Serotonin N-acetyltransferase (AA-NAT; arylalkylamine N-acetyltransferase; EC 2.3.1.87) is a key ... more Serotonin N-acetyltransferase (AA-NAT; arylalkylamine N-acetyltransferase; EC 2.3.1.87) is a key regulatory enzyme in the biosynthesis of melatonin. Previous studies have shown that the activity of this enzyme in the chicken retina is regulated by a cyclic AMP-dependent mechanism. In the present report, we investigated whether cyclic AMP can regulate the levels of AA-NAT mRNA in photoreceptor-enriched chick retinal cell cultures. AA-NAT mRNA levels were elevated by acute treatment with cyclic AMP protagonists, including forskolin; this response was blocked by H-89, a selective inhibitor of cyclic AMP-dependent protein kinase. Forskolin did not alter the rate of disappearance of AA-NAT mRNA in actinomycin D-treated cells, suggesting that cyclic AMP enhances transcription of the AA-NAT gene. Forskolin-induced elevation of AA-NAT mRNA levels was enhanced by cycloheximide, which decreased the degradation of the transcript in cells treated with actinomycin D. These studies indicate that the abundance of AA-NAT mRNA is regulated in part through a cyclic AMP-dependent mechanism.

General and Comparative Endocrinology, 2015

The circadian system drives daily physiological and behavioral rhythms that allow animals to anti... more The circadian system drives daily physiological and behavioral rhythms that allow animals to anticipate cyclic environmental changes. The discovery of the known as ''clock genes'', which are very well conserved through vertebrate phylogeny, highlighted the molecular mechanism of circadian oscillators functioning, based on transcription and translation cycles (24 h) of such clock genes. Studies in goldfish have shown that the circadian system in this species is formed by a net of oscillators distributed at central and peripheral locations, as the retina, brain, gut and liver, among others. In this work we review the existing information about the hepatic oscillator in goldfish due to its relevance in metabolism, and its key role as target of a variety of humoral signals. Different input signals modify the molecular clockwork in the liver of goldfish. Among them, there are environmental cues (photocycle and feeding regime) and different encephalic and peripheral endogenous signals (orexin, ghrelin and glucocorticoids). Per clock genes seem to be a common target for different signals. Thus, this genes family might be important for shifting the hepatic oscillator. The physiological relevance of the crosstalking between metabolic and feeding-related hormones and the hepatic clock sets the stage for the hypothesis that these hormones could act as ''internal zeitgebers'' communicating oscillators in the goldfish circadian system.

Neuroreport, Jan 13, 1999

Tryptophan hydroxylase (TPH) is the first enzyme in the biosynthetic pathways of melatonin in pho... more Tryptophan hydroxylase (TPH) is the first enzyme in the biosynthetic pathways of melatonin in photoreceptor cells and of serotonin in amacrine cells. To assess the regulation of TPH activity in photoreceptor cells, we pretreated retinas with kainic acid. The neurotoxin selectively killed inner retinal neurons while sparing photoreceptors. TPH activity in both control and kainate-treated retinas undergoes a day-night rhythm. The rhythms in both preparations fit sinusoidal functions. However, the rhythm in intact retinas peaks at midday while that in kainate-lesioned retinas does so at midnight. The daily rhythm of tryptophan hydroxylase activity in photoreceptors parallels that of melatonin release. Comparing the mean level of activity in rhythms of intact and lesioned retinas, we calculate that the TPH activity in photoreceptors represents 24% of the total activity. Therefore, the TPH activity measured in intact retinas reflects mainly the enzymatic activity in serotonergic neurons,...

Fat and protein content in relation to age (yearlings and adults), sex and the migratory period (... more Fat and protein content in relation to age (yearlings and adults), sex and the migratory period (autumn migration, winter and spring migration) of the black-headed gull, Larus riübundus have been determined. The omental and subcutaneous fat depots, protein and lipid content of the pectoral muscle, and total plasma lipids were lowest at the end of the autumn migratory period and highest at the vernal migratory period. The fat depots in adults were smaller than in juvenile gulls at the end of the autumn migratory period; however, adults were much fatter in spring. Age-related üfrerences in lipid stores in relation to age-dependent variation in subsequent breeding activity is discussed.

Revista española de fisiología

The present experiments test the effects of intracerebroventricular injections of ovine corticotr... more The present experiments test the effects of intracerebroventricular injections of ovine corticotropin-releasing factor (1 microgram) on food intake, plasma glucose levels and thyroid function at 8 h postinjection in tench. Food intake and thyroid triiodothyronine (T3) content were significantly decreased after CRF treatment. Thyroid thyroxine (T4) content and plasma glucose levels were not modified by this neuropeptide. The present results suggest that CRF plays a role on food intake regulation and thyroid gland activity in tench.

Neurogastroenterology & Motility, 2010

Background Serotonin (5-HT) plays a critical role in several gastrointestinal functions in verteb... more Background Serotonin (5-HT) plays a critical role in several gastrointestinal functions in vertebrates. In teleosts lacking enterochromaffin cells, intestinal 5-HT originates from serotonergic enteric neurons. In the present study, the foregut of a stomachless teleost, the goldfish (Carassius auratus), was used to evaluate the in vitro effect of 5-HT on fish intestinal motility. We also studied the role of melatonin (MEL), an indoleamine sharing the biosynthetic pathway with 5-HT, as regulator of serotonergic activity. Methods An organ bath system, with longitudinal strips from the goldfish intestinal bulb attached to an isometric transducer was used to record foregut smooth muscle contractions. Key Results Concentration-dependent curves of the contractile response exerted by 5-HT and its agonists, 5-methoxytryptamine (5-MT) and 5-carboxamidotryptamine (5-CT), suggest a receptor-mediated action, supported by the blockade by a general 5-HT antagonist, methysergide. The 5-HT-induced contraction was abolished in the presence of atropine, revealing the involvement of cholinergic transmission in gut actions of 5-HT. Furthermore, MEL inhibited the contractile effect of 5-HT and its agonists by up to 50%, which was counteracted by MEL antagonists. Conclusions & Inferences We can provisionally propose that at least two different 5-HT receptor subtypes are involved in fish intestinal motility, a 5-HT 4 -like (5-MTpreferring) and a 5-HT 7 -like (5-CT-and fluphenazinesensitive) receptor. In summary, our results indicate that 5-HT regulates the contractile activity of goldfish foregut through specific receptors located in cholinergic neurons, and that MEL can modulate these serotonergic actions through high-affinity membrane receptors.

Comparative Biochemistry and Physiology Part C: Pharmacology, Toxicology and Endocrinology, 1996

Plasma triiodothyronine (T,) and thyroxine (T4) levels, as well as thyroid free (f) and bound (b)... more Plasma triiodothyronine (T,) and thyroxine (T4) levels, as well as thyroid free (f) and bound (b) thyroid hormones (TH) content, were determined by radioimmunoassay (RIA) in adult Ranu gereri frogs during a 24 h cycle in winter, spring, summer, and autumn. Significant daily changes in plasma Tj levels were present in all the seasons except for winter, being the lowest values observed during the scotophase. In contrast, plasma T4 only showed significant changes in spring, following a similar pattern to the one described for T3.

Physiology & Behavior, 2001

Feeding regulation by monoamines, neuropeptides and certain hormones has been studied in fish, bu... more Feeding regulation by monoamines, neuropeptides and certain hormones has been studied in fish, but a possible role of melatonin is unknown. The purpose of the present study was to investigate the effects of melatonin on food intake in goldfish. Fishes were housed in 12L:12D and injected with different doses of either melatonin or 2-iodomelatonin. Two routes of administration, intracerebroventricular and intraperitoneal injections, and two times of the daily photocycle, midday and midnight, were tested. Food intake was measured at 2, 5 and 8 h postinjection. Melatonin and its analog, 2-iodomelatonin intracerebroventricularly injected had no effect on food intake at any time. However, intraperitoneal injections of both indoleamines significantly reduced food intake at different postinjection times. The inhibitory effect of melatonin was blocked by intraperitoneal administration of its antagonist, luzindole. These results demonstrate the in vivo efficiency of luzindole as melatonin antagonist, and thus provide a useful experimental tool to investigate melatonin functions. In conclusion, both melatonin and its agonist 2-iodomelatonin administered peripherally, inhibit food intake in goldfish, and this inhibitory effect appears to be mediated via luzindole-sensitive melatonin receptors. Our results strongly suggest that melatonin is involved in the peripheral satiety mechanisms in goldfish. D

Pharmacology Biochemistry and Behavior, 1995

In the present study we examined the effects of acute corticotropin-releasing factor (CRF) admini... more In the present study we examined the effects of acute corticotropin-releasing factor (CRF) administration and refeeding treatment on glucose levels and thyroid hormones (plasma levels and thyroid contents) in 48-h food-deprived goldfish. Central CRF administration (2 micrograms) decreased food intake and the thyroid T3 free fraction, without significantly modifying either thyroid hormones bound fractions (T3 and T4) or plasma glucose levels. Subsequently, we tested whether CRF affects thyroid activity by itself, or whether this effect is mediated by CRF-induced feeding reduction. CRF treatment in fasted fish reduced thyroid-free T3 and increased thyroid-free T4, which could be mediated by a decreased intrathyroidal 5&amp;amp;amp;#39;-monodeiodinase activity. These data suggest a CRF effect upon thyroid activity independent of feeding reduction. On the other hand, refeeding after 48-h fasting caused a significant increase in thyroid free T4 content and plasma thyroid hormone levels. Thus, a relationship between nutritional status and thyroid function, which could overlap with CRF effects, cannot be discarded. Plasma glucose levels were only significantly modified by refeeding, which seems to be the signal triggering the increase in glucose titers. Our results support the existence of both CRF-thyroid activity and nutritional status-thyroid function interactions in goldfish.

Pfl�gers Archiv European Journal of Physiology, 1994

The adaptive mechanisms of serotonin N-acetyltransferase (NAT) activity in the regulation of mela... more The adaptive mechanisms of serotonin N-acetyltransferase (NAT) activity in the regulation of melatonin synthesis in frog retina in the face of chronic and acute temperature changes have been investigated. We performed thermal acclimation experiments to test different environmental temperatures at two seasons of the year (summer and winter), followed by the set-up of an eyecup culture system to investigate the acute effects of temperature on NAT activity and melatonin production daily rhythms. Low temperature induced a significant increase in NAT activity, independent of both the time of the photocycle (midday or midnight) and the season of the year (winter or summer). Acute cold-induced stimulation of NAT activity may be associated with lower decreases in the enzyme synthesis rate, rather than decreases in the degradation rate. In contrast, acclimation to warm temperature (25 ~ C) stimulated ocular melatonin production. Nocturnal melatonin production in eyecups cultured at 25 ~ C was significantly higher than in eyecups cultured at 5 ~ C. We suggest ,~hat this discrepancy in thermal regulation of melatonin synthesis can be justified by a seasonal variation in serotonin content within the photoreceptor cells, which determines the thermal response of melatonin production through changes in NAT kinetics.

Neurosignals, 1997

Serotonin N-acetyltransferase (AA-NAT; arylalkylamine N-acetyltransferase; EC 2.3.1.87) is the pe... more Serotonin N-acetyltransferase (AA-NAT; arylalkylamine N-acetyltransferase; EC 2.3.1.87) is the penultimate enzyme in melatonin synthesis and large changes in the activity of this enzyme appear to regulate the rhythm in melatonin synthesis. Recent advances have made it possible to study the mRNA encoding chicken AA-NAT, which has only been detected in the retina and pineal gland. Within the retina, AA-NAT mRNA is expressed primarily in photoreceptors. The levels of chicken retinal AA-NAT mRNA and activity exhibit 24-hour rhythms with peaks at night. These rhythms appear to reflect circadian clock control of AA-NAT mRNA abundance and independent effects of light and darkness on both mRNA levels and enzyme activity. The effects of darkness and light may occur through alterations in cAMP-dependent protein phosphorylation, which increases AA-NAT activity in photoreceptor cell cultures. The cAMP-dependent increase of AA-NAT enzyme activity reflects, at least in part, increased mRNA levels and inhibition of enzyme inactivation by a posttranslational mechanism. This review discusses a hypothetical model for the cellular and molecular regulation of AA-NAT activity by circadian oscillators and light in chicken retinal photoreceptor cells.

Neuroscience Letters, 1993

Temperature effects on ocular serotonin N-acetyltransferase (NAT) kinetics characteristics from R... more Temperature effects on ocular serotonin N-acetyltransferase (NAT) kinetics characteristics from Rana perezi have been studied with respect to tryptamine and serotonin as substrates. Monoamine oxidase (MAO) activity does not interfere in NAT assay at acceptoramine concentrations used in NAT kinetics characterization from R. perezi retina. NAT shows an inhibition by high substrate (serotonin) concentration, which is temperature-dependent. NAT follows the Michaelis-Menten equation at low temperature, whereas at high temperatures (> 10 degrees C) an inhibition by serotonin is observed. This inhibition of NAT activity by serotonin could act as an amplification mechanism to increase daily melatonin rhythm amplitude in the retina of ectotherms.

Neuroscience Letters, 1992

The thermal sensitivity and the response to thermal acclimation of the serotonin N-acetyltransfer... more The thermal sensitivity and the response to thermal acclimation of the serotonin N-acetyltransferase (NAT) activity in frog ocular tissue were studied. The ocular NAT shows a positive thermal modulation for both cosubstrates (tryptamine and acetyl-CoA). The higher NAT activity in the cold-acclimated group (4 degrees C) with respect to the warm-acclimated one (24 degrees C) implies a partial thermal compensation of the enzyme. The present results suggest that frog ocular NAT response to temperature entails a modulation of thermal sensitivity of the enzyme rather than changes in enzyme concentration.

Neuroscience Letters, 2001

Concentration of dopamine (DA) and its metabolite, 3,4-dihydroxyphenylacetic acid is quanti®ed by... more Concentration of dopamine (DA) and its metabolite, 3,4-dihydroxyphenylacetic acid is quanti®ed by high-pressure liquid chromatography with a coulometric detection system in the eye of Xenopus laevis through ontogeny and in adults at two times during photocycle (midday and midnight). Ocular dopaminergic activity remains low during pre-and prometamorphosis and signi®cantly rises in postmetamorphic froglets. This increase is more pronounced at midnight than at midday. The dualism of DA content versus DA release in Xenopus ocular tissue is studied in an eyecup culture system. On a 24-h cycle of DA release from adult Xenopus eyecups the highest DA release by eyecups is produced during daytime, and signi®cantly decreases in darkness. From these results it can be concluded that in spite of the early development of the retinal dopaminergic system in the ontogeny of Xenopus, the ®nal maturation must occur during the metamorphic climax. Endogenous DA release is signi®cantly inhibited by light offset, which explains the higher ocular DA content found at midnight as compared to midday in postmetamorphic froglets and adults. q

Neurochemistry International, 2013

Melatonin has been found in the digestive tract of many vertebrates. However, the enzymatic activ... more Melatonin has been found in the digestive tract of many vertebrates. However, the enzymatic activity of the arylalkylamine-N-acetyltransferase (AANAT) and the hydroxindole-O-methyltransferase (HIOMT), the last two enzymes of melatonin biosynthesis, have been only measured in rat liver. Therefore, the first objective of the present study is to investigate the functionality of these enzymes in the liver and gut of goldfish, analyzing its possible daily changes and comparing its catalytic properties with those from the retina isoforms. The daily rhythms with nocturnal acrophases in retinal AANAT and HIOMT activities support their role in melatonin biosynthesis. In foregut AANAT activity also show a daily rhythm while in liver and hindgut significant but not rhythmic levels of AANAT activity are found. HIOMT activity is not detected in any of these peripheral tissues suggesting an alternative role for AANAT besides melatonin synthesis. The failure to detect functional HIOMT activity in both, liver and gut, led us to investigate other physiological substrates for the AANAT, as dopamine, searching alternative roles for this enzyme in the goldfish gut. Dopamine competes with tryptamine and inhibits retinal, intestinal and hepatic N-acetyltryptamine production, suggesting that the active isoform in gut is AANAT1. Besides, gut and liver produces N-acetyldopamine in presence of acetyl coenzyme-A and dopamine. This production is not abolished by the presence of folic acid (arylamine N-acetyltransferase inhibitor) in any studied tissue, but a total inhibition occurs in the presence of CoA-S-N-acetyltryptamine (AANAT inhibitor) in liver. Therefore, AANAT1 seems to be an important enzyme in the regulation of dopamine and N-acetyldopamine content in liver. Finally, for the first time in fish we found that dopamine, but not N-acetyldopamine, regulates the gut motility, underlying the broad physiological role of AANAT in the gut.

Neurochemistry International, 1995

Arylamine N-acetyltransferase activity (A-NAT: E.C.2.3.1.5) from Rana perezi retina was studied u... more Arylamine N-acetyltransferase activity (A-NAT: E.C.2.3.1.5) from Rana perezi retina was studied using p-phenetidine as specific substrate. Enzyme characteristics and regulation were compared with respect to the arylalkylamine N-acetyltransferase (AA-NAT: E.C.2.3.1.87) from the same tissue. A-NAT activity is distributed in both neural retina and choroid-pigmented epithelium complex, showing a 10-fold higher specific activity in neural retina. In contrast, AA-NAT activity is restricted to neural retina. Subcellular localization in neural retina indicated that both enzymatic activities are in the supernatant fraction (39,000 g, 20 min). p-Phenetidine acetylation was linear as a function of the neural retina amount in the assay (1/16 to 1 retina), and it is insensitive to phosphate buffer pH in the range 6.5-8.4. A-NAT kinetic showed a hyperbolic shape for both cosubstrates. Kinetic constants were KM = 11.2 microM, Vmax = 0.49 nmol/h/mg prot. for p-phenetidine (50 microM acetyl-CoA), and KM = 113.4 microM, Vmax = 3.1 nmol/h/mg prot. for acetyl-CoA (5 mM p-phenetidine). The additivity test for both enzymatic activities in retina homogenates demonstrated that both acceptor amines do not compete for the catalytic sites. Serotonin addition in the assay modifies differentially the kinetic characteristics of both enzymes. Serotonin acted as a strong mixed inhibitor, mainly competitive in nature (competitive Ki = 18.1 microM; non-competitive Ki = 1.9 mM) for AA-NAT. However, it acted as a weak inhibitor with respect to A-NAT, mainly non-competitive, (competitive Ki = 5.7 mM; non-competitive Ki = 8.7 mM).(ABSTRACT TRUNCATED AT 250 WORDS)