Biomedical subjects

M Alonso-Bedate

Publications and source records attributed to M Alonso-Bedate.

At least 37 records · Page 2Linked to original sources

PubMed1995

Differential characteristics and regulation of arylamine and arylalkylamine N-acetyltransferases in the frog retina (Rana perezi).

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)

Animals↗

PubMed1995

Alterations in food intake and thyroid tissue content by corticotropin-releasing factor in Tinca tinca.

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.

Animals↗

PubMed1994

Ontogeny of ocular serotonin N-acetyltransferase activity daily rhythm in four anuran species.

The profile of daily ocular serotonin N-acetyltransferase (NAT) rhythm has been examined during the development of four anuran amphibians in different environments and of varying phylogenetic origins: Xenopus laevis, Discoglossus pictus, Rana perezi, and Bufo calamita. A significant day/night rhythm of ocular NAT with the acrophase at midnight was first detected at stages 43/44 and 25 in X. laevis and R. perezi, respectively. Total enzymatic activity increased with body weight throughout development, although both mean levels and amplitudes of NAT activity per milligram of protein significantly decreased; this attentuation of rhythm was more significant in postmetamorphic froglets, and the acrophase was delayed about 1 hr. There were no significant differences in daily NAT profiles before and after metamorphosis in D. pictus and B. calamita. The amplitude of the NAT rhythm was maximal (> 10-fold nocturnal values) in Xenopus compared with other species (2- to 3-fold).

Animals↗

PubMed1994

Serotonin N-acetyltransferase activity as a target for temperature in the regulation of melatonin production by frog retina.

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 degrees C) stimulated ocular melatonin production. Nocturnal melatonin production in eyecups cultured at 25 degrees C was significantly higher than in eyecups cultured at 5 degrees C. We suggest that 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.

Acclimatization↗

PubMed1993

The inhibition by indoleamines (tryptamine and serotonin) of ocular serotonin-N-acetyltransferase from Rana perezi is temperature-dependent.

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.

Animals↗

PubMed1993

Serotonin N-acetyltransferase (NAT) activity and melatonin levels in the frog retina are not correlated during the seasonal cycle.

The seasonal pattern of daily serotonin N-acetyltransferase (NAT) and melatonin rhythms in the lateral eye of Rana perezi has been characterized to assess any seasonal relationship between enzymatic activity and melatonin production and to evaluate the photothermal responses of such rhythms. In winter there was no daily rhythm either in NAT activity or in melatonin production, although the highest values of NAT activity and lowest melatonin concentrations were measured at this time. Ocular melatonin showed a high-amplitude diurnal rhythm during the summer, whereas a low-amplitude rhythm was observed in spring and autumn. Diurnal melatonin levels were temperature-independent; however, nocturnal melatonin production significantly increased with temperature. Ocular NAT was always higher at lower than at higher temperatures (two- to threefold), both at night and during the day. As suggested for pineal melatonin production in other poikilotherm species, photoperiod controls the duration of the melatonin nocturnal rise, whereas temperature regulates the amplitude of the daily melatonin rhythm in the frog retina. The lack of correlation between daily NAT and melatonin rhythms in the frog retina may result from the high thermal sensitivity of the ocular NAT and a temperature-dependent inhibition substrate of NAT activity.

Animals↗

PubMed1993

Role of corticotropin-releasing factor (CRF) as a food intake regulator in goldfish.

The effect of intraperitoneal or intracerebroventricular corticotropin-releasing factor (CRF) administration on food intake has been studied in goldfish after 24 h of food deprivation. Food intake was evaluated at different time periods after injection, 0-2, 2-8, and 0-8 h. The 1 and 2 micrograms doses of CRF intracerebroventricularly administered induced a reduction in food intake during the first 2 h, although an increased feeding was observed in the next 6 h. The higher dose of CRF used in this study (3.3 micrograms) increased cumulative food intake at 8 h postinjection. However, intraperitoneal injection of 1 microgram CRF did not modify food intake in any of the studied intervals. These results suggest that CRF may play a role in central regulation of feeding behavior intake in goldfish, and show that CRF effects are time- and dose-dependent.

Animals↗

PubMed1992

Thermal sensitivity and effect of temperature acclimation on ocular serotonin N-acetyltransferase activity in Rana perezi.

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.

Acclimatization↗

PubMed1992

Corticotropin-releasing factor stimulates metamorphosis and increases thyroid hormone concentration in prometamorphic Rana perezi larvae.

Attempts to identify a hypothalamic molecule that stimulates thyrotropin (TSH) secretion from amphibian pituitary have been unsuccessful to date. The effects of mammalian (ovine and human) corticotropin-releasing factor (CRF) on the thyroid function of prometamorphic (Taylor & Kollros stages XI-XVII) (Taylor and Kollros, 1946) Rana perezi larvae were studied. Chronic treatments with both ovine and human CRF (oCRF, hCRF) stimulated metamorphosis while delaying larval growth. Chronic hCRF (1 microgram) administration induced 3.2- and 5.3-fold increases in whole body concentration of thyroxine (T4) and triiodothyronine (T3), respectively. In contrast, the 0.5-microgram dose of hCRF stimulated a significant (3.4-fold) increase in whole body concentration of T4 but not of T3. Histological studies of the thyroid gland revealed a 22% increase in the number of follicles per section as a result of the chronic treatment with oCRF (1 microgram). Acute oCRF (2 micrograms) treatment induced a significant increase in T4 concentration at 4 hr (1.3-fold) and 8 hr (2.3-fold) postinjection. T3 concentration was not altered. These results support previous reports and lead us to conclude that a CRF-like peptide, and not TRH, is involved in the regulation of thyroid activity in anuran amphibians during metamorphosis.

Animals↗

PubMed1992

Characterization of serotonin N-acetyltransferase in the lateral eye of the green frog Rana perezi: protective action of EGTA.

The kinetics of serotonin N-acetyltransferase (NAT) from the lateral eye of Rana perezi have been characterized. NAT from ocular tissue reached maximal activity at a phosphate buffer concentration of 250 mM and a pH of 6.5. Reaction linearity was highly conserved within the homogenate fraction range tested (0.033-0.33). The time course of ocular NAT reaction showed a high linearity at 25 and 35 degrees C. Km and Vmax estimations for acetyl-CoA at a 10 mM tryptamine concentration were 63.3 microM and 4.42 nmol/h per eye, respectively. Regardless of the acceptor amine (tryptamine or serotonin), the Km was not affected by the acetyl-CoA concentration (50 or 250 microM), whereas the Vmax was significantly increased at a 250 microM acetyl-CoA concentration. Ocular NAT showed a higher affinity for serotonin (Km = 20.7 microM) than for tryptamine (Km = 48-60 microM); Vmax, however, was similar for both substrates. Acetyl-CoA does not protect ocular NAT; in contrast, the use of EGTA (greater than or equal to 4 mM) in the assay is essential to protect the enzyme because NAT in ocular crude homogenate shows rapid inactivation. This result suggests that intracellular calcium levels are involved in the NAT inactivation mechanisms in frog ocular tissue.

Acetyl Coenzyme A↗

PubMed1992

Role of environmental temperature and photoperiod in regulation of seasonal testicular activity in the frog, Rana perezi.

To analyze the role of environmental temperature and photoperiod in the regulation of the annual testicular cycle in Rana perezi, we performed experiments combining high (25 +/- 1 degrees C) or low (6 +/- 1 degrees C) temperature and different photoperiod regimens (18L:6D, 12L:12D, and 6L:18D (hours light:hours dark)) during three phases of the reproductive cycle: winter stage (December) and prebreeding (February) and postbreeding (May, June) periods. Low temperature and short photoperiod in winter induced the arrest of the maturation phase of spermatogenesis and the activation of primary spermatogonia proliferation and spermiohistogenesis. Rana perezi testis responded to long days stimulus in winter, even at low temperature, with induction of the maturation phase of the cycle. Exposure of male frogs to either high temperature or long photoperiod induced a decrease in testosterone levels in winter. During the prebreeding period, an increase in environmental temperature caused a reduction in testosterone, and a lengthening in photoperiod produced the opposite effect. Photoperiod had no effect on testosterone levels during the postbreeding period, but low temperature increased testosterone plasma levels. These results suggest that both temperature and photoperiod effects can vary seasonally, depending on the phase of the annual reproductive cycle in R. perezi.

Animals↗

PubMed1990

Effects of melatonin on gonadal steroids and glucose plasma levels in frogs (Rana perezi and Rana temporaria).

The effect of melatonin treatment on the Gonosomatic Index (GSI), ovarian germinal epithelium, plasma estradiol and testosterone levels was studied in Rana perezi females in December. No significant changes were observed in GSI, estradiol, and testosterone levels in melatonin treated animals when compared with saline injected controls, but the percentage of previtellogenic follicles decreased in animals treated with melatonin (100 micrograms). The effect of melatonin treatment on glucose level was studied in Rana perezi females in December and Rana temporaria males in February. In Rana perezi no significant differences were observed between melatonin treated and control animals; however, significant reductions by melatonin treatment were obtained in Rana temporaria. The possibility that the inhibitory effect of melatonin can be observed only when gonadal function and metabolism are stimulated by temperature is discussed.

Animals↗

PubMed1990

Response to pinealectomy and blinding in vitellogenic female frogs (Rana perezi) subjected to high temperature in autumn.

The present experiments were carried out to investigate the effects of pinealectomy and bilateral enucleation on the ovarian activity in Rana perezi frogs maintained in 12-h light--12-h dark photoperiod and 20 +/- 1 degrees C during the vitellogenetic growth in late autumn. These environmental conditions, mainly temperature, induce a gonadal and metabolic response similar to that observed in the natural habitat in summer: a marked ovarian follicular regression, a depletion of the energetic resources from fat bodies and liver, and a minimum in oestradiol circulating levels. This response is partially blocked by pinealectomy and blinding. Protein phosphorus, as an index of vitellogenic proteins, and total ovary lipid content were significantly higher in pinealectomized and blinded frogs with respect to sham-operated animals. Likewise, oestradiol concentrations showed a significant increase during the dark phase of the daily photocycle in pinealectomized and blinded animals. From our results, we can suggest that the arrest of vitellogenesis, the depletion of energetic resources, and the regulation of oestradiol levels induced by the high temperature in Rana perezi frogs can be influenced, at least in part, by the pineal complex and lateral eyes.

Adipose Tissue↗

PubMed1989

Seasonal cycles in testicular activity in the frog, Rana perezi.

Studies of seasonal testicular cycle based on spermatogenetic activity and direct measurement of plasma testosterone were made in male frog Rana perezi obtained from its natural biotope in the Iberian Peninsula. Testosterone plasma level was determined by radioimmunoassay and exhibited notable differences according to season: plasma testosterone was lowest (less than 0.5 ng/ml) in summer and then increased progressively to reach a peak in spring (3-4 ng/ml), coincident with mating. After spermiation, when an increase in temperature and photoperiod in the natural habitat occurs, levels decline. Fat bodies also show a pronounced seasonal cycle with total regression following breeding and maximal development in winter. However, testicular weight was independent of seasons, and no significant change was observed throughout the year. Histological evidence indicates that although cell nests of different types are present every month of the year, the most important spermatogenetic activity is initiated in summer. The possible relationship between spermatogenetic activity and testosterone production and the importance of environmental factors as synchronizers of seasonal reproduction are discussed.

Animals↗

PubMed1988

In vivo effect of melatonin and gonadotropin-releasing hormone on testicular function in Rana temporaria.

Studies concerning pineal-gonadal interactions and investigations about the effect of the putative pineal hormone melatonin in male amphibians are very scarce. In this work we investigate the effects of melatonin, synthetic gonadotropin-releasing hormone (GnRH), or both when injected in Rana temporaria males. We conclude that melatonin inhibits spermiation and spermatogonial multiplication when injected alone or with GnRH. We have been unable to find any significant differences in plasma testosterone levels when melatonin or GnRH are injected alone, but when melatonin is injected with GnRH the levels of testosterone are significantly increased. These results seem to demonstrate that melatonin could play a role in the regulation of the gonadal function in the male Rana temporaria.

Animals↗

PubMed1988

Oxygen regulation capacity in Discoglossus pictus tadpoles between moderate hyperoxia and acute hypoxia in water.

The oxygen dependence of aquatic oxygen consumption was measured in active and anesthetized stage XVIII Discoglossus pictus tadpoles (Amphibia, Anura). The active tadpoles are good oxygen regulators in moderate hyperoxia and moderate hypoxia, whereas they are oxygen conformers in acute hypoxia. Critical oxygen pressure was 52 mmHg O2. Anesthetizing the larvae changes them to perfect oxygen conformers between moderate hyperoxia and moderate hypoxia (249-63 mmHg O2). At stage XVIII the aquatic respiratory organs are still capable of producing oxygen regulation when free access to air is denied. The marked capacity for oxygen regulation in D. pictus tadpoles is concordant with the strong hypoxic environments in which these animals usually live in nature.

Animals↗

PubMed1987

Different levels of hyperoxia reversibly induce catalase activity in amphibian tadpoles.

Studies about the proposed antioxidant physiological role of the catalase (CAT) enzyme in relation to different environmental oxygen tensions are reported for the first time in amphibian larvae of Discoglossus pictus and Rana ridibunda perezi during their development. The CAT levels of whole tadpoles increased constantly in both species during the larval period, reaching a maximum during the metamorphic climax. All through development, CAT activity levels were always greater in D. pictus than in R. ridibunda perezi. This correlates well with the already reported higher SOD activity and hyperoxia resistance of the D. pictus species when compared to R. ridibunda perezi. Long-term acclimation to different levels of hyperoxia (40, 60, and 100% O2) showed dose-related increases in the CAT activity of D. pictus tadpoles. These increases did not take place when the animals were subjected to acute hyperoxia (24 h). The increase in CAT activity observed after 15 days of acclimation to acute hyperoxia (710 mm Hg: 100% O2) was reversed after 15 additional days of postacclimation to normal air (149 mm Hg O2). When recently metamorphosed frogs were acclimated to acute hyperoxia, significant increases in CAT activity were observed after 15 days, but not after 7 days. The results are interpreted as supporting a protective role for the CAT enzyme in amphibian larvae and froglets against oxygen toxicity.

Aging↗

PubMed1985

Catalase enzymatic activity and electrophoretic patterns in adult amphibians--a comparative study.

Catalase electrophoretic patterns and enzymatic activities were measured in four organs of two anuran species, Rana ridibunda perezii and Discoglossus pictus. The D. pictus enzyme appeared as two distinguishable bands, whereas R. ridibunda catalase was monomorphic. Electrophoretic mobility of the major D. pictus catalase band was greater than that of R. ridibunda. Enzymes from both species showed slower mobility than that from bovine liver. Catalase activities did not show significant differences according to sex in any of the organs tested in R. ridibunda. Enzyme activities were similar in liver, kidney and brain when both species were compared. Only the heart showed much higher activity in D. pictus than in R. ridibunda. The catalase activity levels followed the order: liver greater than kidney greater than heart in both species. The heart showed higher activity than the brain in D. pictus but not in R. ridibunda.

Animals↗