Biomedical subjects

L G Branco

Publications and source records attributed to L G Branco.

14 recordsLinked to original sources

PubMed1997

Effects of 2-deoxy-D-glucose and insulin on plasma glucose levels and behavioral thermoregulation of toads.

The present study was designed to test the hypothesis that hypoglycemia induces hypothermia in ectotherms and to elucidate the mechanisms responsible for behavioral hypothermia. Behavioral hypothermia is a stress response that occurs in organisms ranging from protozoans to mammals, but very little is known about the cellular mechanisms involved. Toads equipped with a temperature probe were tested in a thermal gradient (10-40 degrees C). Insulin was used to reduce plasma glucose levels, and an inhibitor of glucose utilization, 2-deoxy-D-glucose (2-DG), was used to cause intracellular glucopenia. Insulin injections into the dorsal lymph sac caused significant reductions of both plasma glucose levels and body temperature. To determine if the response was mediated by extracellular glucose receptors or an intracellular mechanism. 2-DG was also injected into the lymph sac. 2-DG caused a similar drop in body temperature and a marked increase in plasma glucose. To assess the role of central thermoregulatory mechanisms, a smaller dose of 2-DG was injected into the fourth cerebral ventricle or the lymph sac. Intracerebroventricular injection of 2-DG caused a decrease in body temperature despite elevated circulating glucose levels, whereas injection into the lymph sac caused no significant change. The data indicate that exclusion of glucose from central rather than peripheral sites plays a major role in the hypoglycemia-induced behavioral hypothermia and that intracellular mechanisms rather than extracellular glucose receptors are involved in this response. Hypothermia may be a beneficial response to hypoglycemia in toads because it dampens cellular oxidative demands during glucose deprivation.

Animals↗

PubMed1996

Thermoregulatory effects of cyanide and azide in the toad, Bufo marinus.

An important adaptation to hypoxia is a regulated reduction in body temperature (Tb; anapyrexia), presumably because it lowers metabolic rate when oxygen supply is limited. Although this beneficial response occurs in organisms ranging from protozoans to mammals, little is known of the cellular mechanisms involved. We showed previously that inhibition of oxidative phosphorylation mediates hypoxia-induced anapyrexia in the paramecium. In this study, we tested the hypothesis that inhibition of oxidative phosphorylation also causes anapyrexia in a vertebrate, Bufo marinus. Tb in toads was measured in a thermal gradient 24 h before and 24 h after administration of either NaCN or NaN3, both inhibitors of oxidative phosphorylation. Subcutaneous NaCN (0.6 mmol/kg) reduced Tb from 29.1 +/- 0.8 to 19.6 +/- 0.6 degree C (P = 0.002). Infusion of NaCN (0.6 mumol/kg) into the fourth ventricle of the brain reduced Tb from 30.0 +/- 0.9 to 24.8 +/- 1.2 degrees C (P = 0.01). Responses to NaN3 were similar to the NaCN responses. Control injections and subcutaneous injections of 0.6 mumol/kg NaCN and NaN3 had no significant effect on Tb (P > 0.32). Neither NaCN nor NaN3 had significant effects on arterial PO2, PCO2, or pH at 26 degrees C. These results indicate that inhibition of oxidative phosphorylation in the central nervous system leads to the selection of cooler temperatures. Thus reduced oxidative phosphorylation within the brain may be an important factor eliciting hypoxia-induced anapyrexia.

Animals↗

PubMed1996

Characterization of the atrial natriuretic factor system in lungs of the toad Bufo paracnemis.

Blood pressure in the amphibian pulmonary circulation is relatively high because a single ventricle serves both the systemic and pulmonary circulation, creating a high degree of plasma filtration from pulmonary capillaries. Previous studies have shown that lung atrial natriuretic factor (ANF) may have an important physiological function in preventing edema in mammals. In this study, we report the presence of the complete ANF system in the lungs of the toad Bufo paracnemis. Radioimmunoassay of tissue homogenates revealed that toad lung ANF concentration was approximately twice as high (928.5 +/- 83.0 pg mg-1 protein) as that of lung tissue in mammals of a similar size. The amount of ANF was significantly higher in the left than in the right atrium (15.0 +/- 1.2 versus 1.9 +/- 0.8 ng mg-1 protein; N = 4, P < 0.001), while the ventricle contained 488.3 +/- 41.8 pg mg-1 protein. In extracts of both lungs and atria, high-performance liquid chromatography revealed two forms of the peptide; prohormone and a carboxy-terminal peptide of low molecular mass, which is the biologically active form of peptide. The presence of the prohormone suggests that ANF is synthesized in toad lungs and atria. Characterization of toad lung receptors by a competitive binding assay demonstrated three different subtypes of ANF receptors: the guanylyl cyclase (GC) receptors, GC-A and GC-B, as well as clearance (C) receptors. We conclude that the toad Bufo paracnemis has a well-developed complete ANF system in the lung, suggesting that it has a role in toad lung physiology.

Amino Acid Sequence↗

PubMed1995

Ventilatory responses to carboxyhaemoglobinaemia and hypoxic hypoxia in Bufo paracnemis.

The modality of the O2 drive to breathe was evaluated in unanaesthetized Bufo paracnemis. Carbon monoxide (CO) hypoxia was applied to reduce CaO2 (arterial O2 content). Arterial PO2 (PaO2) was reduced through inspiration of hypoxic gas mixtures (hypoxic hypoxia). Measurements included pulmonary ventilation, CaO2, PaO2, pHa, blood pressure and heart rate. Application of fractional concentrations of CO equivalent to 0.001 reduced CaO2 from 11.6 +/- 1.2 to 8.6 +/- 0.7 vol% (mean +/- S.E.M., N = 6) without any effect on heart rate, blood pressure or ventilation. Inspiration of fractional concentrations of CO equivalent to 0.01 reduced CaO2 to 5.1 +/- 0.7 vol%, which was accompanied by increased ventilation. This response, however, correlated with a decreased arterial pH. The effect of a fractional concentration of CO of 0.01 on ventilation was abolished by compensation of pH through bicarbonate infusion. Consequently, carboxyhaemoglobinaemia per se failed to increase ventilation. By contrast, reductions of PaO2 clearly stimulated ventilation, which indicates that O2 partial pressure provides the principal O2 stimulus to breathe in Bufo paracnemis.

Animals↗

PubMed1994

Role of central chemoreceptors in behavioral thermoregulation of the toad, Bufo marinus.

We tested the hypothesis that hypercapnia will induce behavioral hypothermia in toads and that central chemoreceptors are involved in this response. Animals were tested in an enclosed temperature gradient supplied with different gas mixtures. Fractional inspired CO2 (FICO2) between 0 and 0.05 had no significant effect on selected body temperature, but FICO2 between 0.06 and 0.10 reduced the selected body temperature from U approximately 28 to 18 degrees C. To determine if the hypercapnia-induced hypothermia is mediated by acidification of central chemoreceptors, the pH of the fourth ventricle was kept constant by perfusion with mock cerebrospinal fluid of pH 7.7 or 7.1 (normal and acidic values, respectively). Ventricular perfusion at pH 7.7 under normocapnic conditions had no effect on body temperature. Hypercapnia (FICO2 0.08) failed to induce hypothermia when the fourth ventricle was kept at pH 7.7 and when hyperoxia was present. Acidic ventricular perfusion under normocapnic conditions decreased selected body temperature from 27 to 25 degrees C, a significant drop but much less than that due to hypercapnia producing the same brain pH, suggesting an important role of peripheral chemoreceptors. The physiological significance of behavioral hypothermia and nature of the peripheral stimulus were evaluated by measuring the effect of hypercapnia on arterial oxygen saturation, PO2, and pH at 15 and 25 degrees C. Arterial oxygen saturation was higher at the lower temperature. Increasing FICO2 decreased oxygen saturation at 25 degrees C but not at 15 degrees C. Arterial PO2 increased with increasing inspired CO2. This increase was greater at 15 degrees C than at 25 degrees C. Arterial pH decreased at both temperatures.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1994

A new function for lactate in the toad Bufo marinus.

In the amphibian Bufo marinus, progressive hypoxia below a critical PO2 elicits a transient 50% increase in O2 consumption that coincides with the onset of lactate formation. The present study was designed to test the hypothesis that lactate causes the observed rise in metabolic rate. Arterial bolus infusions of pH-neutral sodium lactate solutions (4 mmol/kg body wt) in toads maintained under hypoxia actually elicit a similar increase in metabolic rate. The application of adrenergic antagonists (bretylium tosylate, phentolamine, propranolol, and reserpine) inhibits this response, suggesting that catecholamines are involved. Moreover, animals injected with lactate move to a cooler environment (behavioral hypothermia), a behavioral response that is beneficial during hypoxia. We hypothesize that, in accordance with Cannon's concept of an emergency response, lactate may function as an alarm signal during hypoxia. However, the signal function of lactate is observed in animals both under hypoxia and under normoxia and should thus be considered in future studies whenever elevated lactate levels are present, e.g., during and after exercise.

Alkalosis, Respiratory↗

PubMed1994

Ventilatory responses to hypoxia in the toad Bufo paracnemis before and after a decrease in haemoglobin oxygen-carrying capacity.

The ventilatory and cardiovascular effects of a decreased O2-carrying capacity of the blood were evaluated in the toad Bufo paracnemis. Pulmonary ventilation was monitored using a pneumotachographic technique. Chronic arterial catheters served to record both cardiac frequency and blood pressure and enabled the withdrawal of blood samples for analysis of pH and partial pressure of O2 (PO2). Haemoglobin concentrations were determined by the cyanmethaemoglobin method. The ventilatory response to hypoxia was not affected by the reduction in blood O2-carrying capacity, which suggests that PO2 rather than O2 content is the regulated variable. The reduction in capacity is accompanied by an increased normoxic heart rate and by a reversal of the hypoxic tachycardia normally observed.

Animals↗

PubMed1993

Temperature and central chemoreceptor drive to ventilation in toad (Bufo paracnemis).

The central chemoreceptor drive to ventilation was assessed in unanesthetized toads, Bufo paracnemis, exposed to three different temperatures: 15, 25 and 35 degrees C. The acid-base status of the fourth ventricle was manipulated by mock CSF perfusion. In additional experiments, arterial pH was varied by inspiration of hypercapnic gas mixtures. Ventilation was measured directly by pneumotachography and arterial blood samples were analyzed using electrodes for pH and PO2. Regardless of temperature, the ventilatory control of acid-base status was predominantly central. Moreover, an increase in temperature was accompanied by a proportional increase in the ventilatory response to chemoreceptor stimulation by either lowered mock CSF pH or hypercapnia. The alphastat hypothesis could not adequately account for the temperature effects on the ventilatory responses to hypercapnia or on air convection requirements in the toad.

Animals↗

PubMed1993

Interaction between temperature and hypoxia in the alligator.

Hypoxia elicits behavioral hypothermia in alligators. Under normoxic conditions, the selected body temperature is 27.8 +/- 1.2 degrees C. However, when inspired O2 is lowered to 4%, selected body temperature decreases to 15.4 +/- 1.0 degrees C. The threshold for the behavioral hypothermia is between 4 and 5% inspired O2, the lowest threshold measured so far in terrestrial vertebrates. This study assessed the physiological significance of the behavioral hypothermia. The body temperature was clamped at 15, 25, and 35 degrees C for measurements of ventilation, blood gases, metabolic rate, plasma lactate, and acid-base status. Hypoxia-induced changes in ventilation, acid-base status, oxygen consumption, and lactate were proportional to body temperature, being pronounced at 35 degrees C, less at 25 degrees C, and absent at 15 degrees C. The correlation between selected body temperature under severe hypoxia and the measured parameters show that behavioral hypothermia is a beneficial response to hypoxia in alligators.

Alligators and Crocodiles↗

PubMed1993

Effect of temperature on central chemical control of ventilation in the alligator Alligator mississippiensis.

Central chemoreceptor function was assessed in unanesthetized alligators, Alligator mississippiensis, at body temperatures of 15, 25 and 35 degrees C. Two experiments were performed. In the first experiment, the fourth ventricle was perfused with mock cerebrospinal fluid (CSF) solutions of different pH values (7.1-7.9). Changes in pulmonary ventilation were evaluated with a pneumotachograph and arterial pH (pHa) was measured. Perfusion with low-pH solutions increased ventilation and arterial pH. Perfusion with high-pH solutions decreased ventilation and arterial pH. Mock CSF pH had a greater effect at higher temperatures. In the second experiment, the relative contributions of central and peripheral chemoreceptor drive to breathing were evaluated using hypercapnic gas mixtures to stimulate both central and peripheral chemoreceptors. Hypercapnia caused an increase in ventilation which was larger at higher temperatures. To stimulate only the peripheral chemoreceptors, the same hypercapnic gas mixtures were applied while the CSF pH of the fourth ventricle was kept constant by perfusion with a mock CSF solution. This reduced significantly the ventilatory response induced by hypercapnia. These data indicate that, regardless of the temperature, central chemoreceptors play a major role in the ventilatory regulation of the alligator. The change in pHa with temperature is compatible with the alphastat hypothesis.

Alligators and Crocodiles↗

PubMed1992

Toad bladder amiloride-sensitive channels reconstituted into planar lipid bilayers.

In the present study we used established methods to obtain apical membrane vesicles from the toad urinary bladder and incorporated these membrane fragments to solvent-free planar lipid bilayer membranes. This resulted in the appearance of a macroscopic conductance highly sensitive to the diuretic amiloride added to the cis side. The blockage is voltage dependent and well described by a model which assumes that the drug binds to sites in the channel lumen. This binding site is localized at about 15% of the electric field across the membrane. The apparent inhibition constant (K(0)) is equal to 0.98 microM. Ca2+, in the micromolar range on the cis side, is a potent blocker of this conductance. The effect of the divalent has a complex voltage dependence and is modulated by pH. At the unitary level we have found two distinct amiloride-blockable channels with conductances of 160 pS (more frequent) and 120 pS. In the absence of the drug the mean open time is around 0.5 sec for both channels and is not dependent on voltage. The channels are cation selective (PNa/PCl = 15) and poorly discriminate between Na+ and K+ (PNa/PK = 2). Amiloride decreases the lifetime in the open state of both channels and also the conductance of the 160-pS channel.

Amiloride↗

PubMed1992

Central chemoreceptor drive to breathing in unanesthetized toads, Bufo paracnemis.

Central chemoreceptor drive to breathing was studied in unanesthetized toads, equipped with face masks to measure pulmonary ventilation and arterial catheters to analyze blood gases. Two series of experiments were performed. Expt. 1: The fourth cerebral ventricle was perfused with solutions of mock CSF, adjusted to stepwise decreasing pH values. Concomitant perfusion-induced increases of pulmonary ventilation, pHa and PaO2 were measured. Expt. 2: Inspiration of hypercapnic gas mixtures was applied to stimulate both central and peripheral chemoreceptors. Subsequently, only peripheral chemoreceptors were stimulated. This was accomplished by repeating the hypercapnic conditions while the fourth ventricle was perfused with mock CSF at pH 7.7. This procedure reduced the slope of the ventilatory response curve by about 80%. Taken together, the experiments suggest a highly dominant role of central chemoreceptors in the ventilatory acid-base regulation of the toad.

Animals↗

PubMed1992

Autonomic basis for hypoxia-induced hyperglycaemia in toads (Bufo paracnemis).

1. Toads were exposed to steady hypoxic conditions (inspired PO2 = 40 mmHg) for 60 min. 2. Within the exposure time, glucose concentrations rose from about 30 mg% to a steady level of 45 mg%. The development of hyperglycaemia reached a stable level within 40 min. 3. This effect was eliminated by treatment with either propranolol or atropine, suggesting combined cholinergic and adrenergic activation.

Animals↗

PubMedDate not supplied

Interactions between body temperature regulation and blood acid-base status in anuran amphibians.

In vertebrate evolution, the transition from aquatic to terrestrial mode of life was associated with considerable changes in the respiratory system and CO2/pH-sensitive receptors became fundamental. The present review focuses on the combined effects of hypercapnia and body temperature in anuran amphibians, that represent a key group for the transition. Recent studies have indicated that temperature affects the hypercapnic drive to breathe. Conversely, hypercapnia modulates the range of preferred body temperature of amphibians and central (CO2/pH) receptors are likely to be involved.

Acid-Base Equilibrium↗