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Biomedical subjects

B Gancedo

Publications and source records attributed to B Gancedo.

13 recordsLinked to original sources

Changes in glucose, glycogen, thyroid activity and hypothalamic catecholamines in tench by starvation and refeeding.

The effects of short-term food deprivation (7 days) and refeeding (2 days) on different biochemical and neuroendocrine parameters were studied in tench. A 7-days fast resulted in a significant reduction of plasma glucose and glycogen hepatic content, supporting the key role of liver glycogen as energy depot for being consumed during fasting. The rapid recovery of normal values of blood glucose and glycogen stores by refeeding indicates a rapid replenishment of liver glycogen stores. The short-term starvation decreased circulating thyroid hormones (both T3 and T4) and T4 release from thyroid, supporting an interaction between nutritional state and thyroid function in tench. All these metabolic and hormonal changes were partial or totally reversed under refeeding conditions. An increase in hypothalamic content of norepinephrine and dopamine was found in fasted fish. This result might be a consequence of stress induced by starvation.

Analysis of Variance↗

Changes in thyroid hormone concentrations and total contents through ontogeny in three anuran species: evidence for daily cycles.

Three anuran species (Rana perezi, Xenopus laevis, and Bufo calamita) of different phylogenetic origins and ecological habitats have been studied during ontogeny with respect to day/night changes in whole-body concentrations and total content of extrathyroidal thyroxine (T4) and triiodothyronine (T3). There were no significant day/night changes in thyroid hormones (TH) during embryonic stages. Daily cycles in TH with higher nocturnal values appeared during premetamorphosis in R. perezi and X. laevis. Cyclicity disappears for T3, while it is reversed for T4, in prometamorphic R. perezi and X. laevis. In contrast, there were significantly higher T3 (0.74 +/- 0.13 ng/g) and T4 (2.08 +/- 0.54 ng/g) levels at night in prometamorphic B. calamita. Significant daily changes in T3 and T4 with higher nocturnal values (T3, 788.29 +/- 118.38 pg/g; T4, 1.95 +/- 0.4 ng/g) were again seen in X. laevis at the end of climax, while in B. calamita low TH values appeared at early scotophase and there were no significant changes in R. perezi at this time. Similar daily profiles were observed for TH whole-body concentrations and total contents.

Animals↗

Effect of alpha-helical-CRF[9-41] on feeding in goldfish: involvement of cortisol and catecholamines.

The anoretic effect of corticotropin-releasing factor (CRF) was not dependent on adrenal activation in goldfish (Carassius auratus). Moreover, an interaction between CRF and the hypothalamic catecholaminergic system in the central regulation of food intake was observed. The intracerebroventricular (icv) administration of CRF increased cortisol levels and reduced food intake and hypothalamic norepinephrine and dopamine content at 2 hr postinjection, with these effects reversed by alpha-helical CRF[9-41] pretreatment. The anoretic effect of CRF was independent of the circulating cortisol increase, because it was only evoked after icv injections but not after intraperitoneal (ip) administration. Furthermore, the increase in plasma cortisol levels induced by ip administration of this steroid did not modify feeding.

Animals↗

Daily changes in thyroid activity in the frog Rana perezi: variation with season.

Plasma triiodothyronine (T3) and thyroxine (T4) levels, as well as thyroid free (f) and bound (b) thyroid hormones (TH) content, were determined by radioimmunoassay (RIA) in adult Rana perezi frogs during a 24 h cycle in winter, spring, summer, and autumn. Significant daily changes in plasma T3 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. Thyroid fT3 content did present day/night significant changes only in spring showing high contents at early scotophase. Mean fT4 content was higher at the beginning of light phase than during the rest of daily photocycle in spring and autumn, but significant differences appeared only in autumn. Regarding the thyroid bound content of TH, bT3, and bT4 presented significant daily changes in spring and autumn. However, different profiles were observed in these two seasons. High bound contents were found at early photo- and scotophase with lower values at late dark phase in spring, whereas higher contents were detected at this time in autumn. The present results indicate the existence of seasonally changing daily fluctuations in thyroid activity in Rana perezi and it seems that an interaction between photoperiod and temperature plays a role in the regulation of these daily changes.

Analysis of Variance↗

Seasonal changes in thyroid activity in male and female frog, Rana perezi.

Plasma T3 and T4 levels, and thyroid free (f) and bound (b) thyroid hormones contents, were determined by radioimmunoassay in adult male and female Rana perezi over 1 year. Plasma T3 levels show a significant seasonal cycle in both male and female frogs, increasing from January to peak in July (71-74 pg/ml) to decrease in August-October. Plasma T4 concentrations also varied seasonally, and sex differences were apparent. Basal T4 levels from the end of summer to the end of winter and higher values (682-651 pg/ml) in spring and early summer occur in female frogs; males have lowest levels in October (59.78 +/- 13.92 pg/ml) and higher levels during the rest of the year, except for a significant decrease in April. The thyroid content of fT3 and fT4 displayed similar seasonal patterns, with peak values in April and July-August. Sex differences are again present. The thyroid contents of bT3 are high between March and July in both sexes, with values in females being greater than those in males. The bT4 content follows a similar seasonal pattern in males and females and shows a minimum in March and a maximum in summer. Thyroid releasing capacity seems to change depending on the season and sex.

Animals↗

CRF effect on thyroid function is not mediated by feeding behavior in goldfish.

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'-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.

Animals↗

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↗

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↗

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↗

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↗

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↗

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↗

Transforming growth factor-beta promotes differentiation of ovarian thecal-interstitial cells but inhibits androgen production.

Evidence that transforming growth factor-beta (TGF beta) is produced by thecal-interstitial cells (TIC) has suggested the hypothesis that TGF beta may be an autocrine regulator of TIC function. The purpose of these studies is to begin to test this hypothesis. In the present experiments we tested the effects of TGF beta on steroid production by TIC isolated from the ovaries of hypophysectomized immature rats by Percoll gradient centrifugation. When TIC (10(4) viable cells/well) were cultured in serum-free medium (0.2 ml in 96-well plates), low amounts of androsterone (less than 4 ng/ml) were produced at 2, 4, and 6 days. TGF beta (0.01-100 ng/ml) did not change basal androsterone production. Treatment with LH (50 ng/ml) stimulated a 100-fold increase in androsterone at 2 days and 60-fold increases at 4 and 6 days. Concomitant treatment with TGF beta (10 ng/ml) caused a 65% inhibition (ED50 = 2.3 +/- 0.7 ng/ml) of androsterone production at each time period. Analysis of key steroid metabolites demonstrated that androsterone and androstenedione were inhibited equally, while progesterone was significantly increased (ED50 = 1.2 +/- 0.2 ng/ml). Time-course studies revealed that TGF beta alone did not alter progesterone production at 2 days, but markedly increased progesterone (10-fold) above control levels at 4 and 6 days. Dose-response experiments showed that TGF beta did not alter the sensitivity of the TIC to LH stimulation, indicating that LH activation of the intracellular signaling pathway was not blocked by TGF beta. Treatment with insulin-like growth factor-I (IGF-I) together with LH caused a synergistic increase in androsterone production. The synergistic stimulation of LH action by IGF-I could be blocked by TGF beta. Interestingly, TIC were more sensitive to TGF beta in the presence of IGF-I (ED50 = 0.18 +/- 0.04 ng/ml). In contrast, TGF beta enhanced progesterone production only at the highest dose of TGF beta (10 ng/ml). To further elucidate the mechanism of TGF beta action, the effects of TGF beta on the TIC content of 17 alpha-hydroxylase/C17-20 lyase (P450(17)alpha) and cholesterol side-chain cleavage (P450scc) were analyzed by immunoblotting. TGF beta alone or in combination with LH stimulated an increase in P450scc content, but did not alter P450(17 alpha content. These results lead us to conclude that 1) the TIC are targets for TGF beta; 2) IGF-I increases the sensitivity of TIC to TGF beta action; and 3) TGF beta acts directly on TIC to stimulate progesterone while inhibiting androgen production.

Androgens↗