Effect of gender on thermoregulation and survival of hypoxic rats.

1. Hypothermia is a documented response to hypoxia but little is known about possible gender differences. Because female rats have a greater hypoxic ventilatory response than males, we hypothesized that females would be more tolerant of hypoxia. We studied 18 female and 18 male Long-Evans rats. 2. Radiotelemetry transmitters for body temperature (Tb) were implanted under general anaesthesia (90 mg/mL ketamine and 10 mg/mL xylazine; 0.1 mL/100 g bodyweight, i.p.). 3. Rats were exposed to 21, 16, 12, 10, 8, 6, 4 and 2% O2 (balance N2) for 30 min each in chambers kept at either 31 degrees C (clamped) or 20 degrees C (hypothermic). Survival was defined as ataxic and unresponsive. 4. Females were more hypoxia tolerant than males, often enduring 2% inspired O2 (13 km altitude). 5. This was correlated with a lower Tb in the hypothermic group, but not in the clamped group. 6. Hypothermia increased 'survival' of rats independent of gender. 7. When Tb was clamped, female rats showed significantly greater survival than males. 8. Thus, separate mechanisms (hypothermia or ventilation) may be acting to increase tolerance of clamped and hypothermic female rats to severe hypoxia.

Physiological characterization of pulmonary carbonic anhydrase in the turtle.

Direct measurements have found that ectothermic vertebrates possess a significant postcapillary PCO2 disequilibrium between arterial blood and alveolar gas, indicating that the CO2-HCO3(-)-H+ system does not reach equilibrium during pulmonary capillary transit. One plausible explanation for the blood disequilibrium is that turtle lungs lack vascular carbonic anhydrase (CA) to enhance the conversion of blood HCO3- to CO2. The present study characterized the contribution of pulmonary vascular CA to CO2 excretion and postcapillary CO2-HCO3(-)-H+ equilibration in the turtle. In situ perfusion of turtle lungs with salines containing membrane-permeating and membrane-impermeant CA inhibitors produced significant and comparable postcapillary pH and PCO2 perfusate disequilibria. Replacement of perfusate chloride with various anions had no affect on pulmonary CO2 excretion, thereby ruling out a significant contribution of Cl- sensitive CA isozymes (i.e., CA II-like). Perfusion of lungs with control salines following treatment with phosphatidylinositol specific-phospholipase C produced significant CO2 disequilibria, consistent with connection of CA IV to the luminal membrane of endothelial cells via a phosphatidylinositol glycan linkage. Vascular CA IV in the turtle lung would participate in diffusive and reactive CO2 equilibration and, thus, may compensate for the slow rate of the physiological anion shift in turtle erythrocytes (Stabenau et al., 1991) during capillary transit.

Anesthetic and postanesthetic management of sea turtles.

OBJECTIVE: To examine the physiologic effects of inhalation anesthesia in aquatic turtles to improve anesthetic techniques and postanesthetic monitoring. DESIGN: Retrospective case series. ANIMALS: 9 Kemp's ridley sea turtles. PROCEDURE: Isoflurane was used as the general anesthetic during 14 minor surgical procedures. Turtles were orotracheally intubated, and a surgical plane of anesthesia was maintained with 2.7 +/- 0.4% (mean +/- SE) isoflurane. The duration of anesthesia was 131 +/- 12 minutes. Pulse rate, blood pressure, blood gases (PaO2 and PaCO2) and pH, blood lactic acid concentration, and capnography were used to evaluate the physiologic responses of sea turtles to isoflurane. RESULTS: An isoflurane concentration of 3.4 +/- 0.3% provided anesthetic induction in 7 +/- 1 minutes. The mean duration of the recovery phase was 241 +/- 31 minutes. The duration of the recovery phase was not affected by the duration of anesthesia, type of carrier gas, method of ventilatory weaning, or use of selected pharmacologic agents. The recovery phase was characterized by hypoxemia, progressive acidemia, hypercapnia, and lactic acidosis. Awakening in the turtles was preceded by a characteristic tachycardia and tachypnea. All sea turtles recovered from isoflurane anesthesia without apparent adverse effects within 24 hours. CLINICAL IMPLICATIONS: Isoflurane appears to be safe and effective in providing surgical anesthesia in turtles that require a timely return to an aquatic environment. This study should assist veterinarians in predicting the physiologic responses of aquatic turtles to inhalation agents.

Roles of intra- and extracellular carbonic anhydrase in alveolar-capillary CO2 equilibration.

Alveolar-capillary CO2 equilibration involves diffusive equilibration of CO2 across the blood-gas barrier and chemical equilibration of perfusate CO2-HCO-3-H+ reactions. These processes are governed by different, but related, driving forces and conductances. The present study examined the importance of pulmonary carbonic anhydrase (CA) for diffusive and reactive CO2 equilibration in isolated rat lungs. Lungs were perfused with salines containing membrane-impermeant or -permeant inhibitors of CA. Measurements of CO2 excretion rate, equilibrated venous and arterial PCO2 and pH, and postcapillary pH and PCO2 disequilibria were used, together with our previous model of CO2-HCO-3-H+ reactions and transport in saline-perfused capillaries (Bidani et al. J. Appl. Physiol. 55: 75-83, 1983), to compute the relevant driving forces and conductances. Reactive CO2 equilibration was markedly affected by extracellular (vascular) CA activity but not by the activity of intracellular (cytosolic) CA. The driving force for CO2 diffusion was strongly influenced by vascular CA activity. The conductance for CO2 diffusion was independent of CA activity. The minimum conductance for CO2 diffusion was estimated to be 700-800 ml.min-1.Torr-1. The results indicate that extracellular vascular CA activity influences both diffusive and reactive CO2 equilibration. However, cytosolic CA has no detectable role in alveolar-capillary CO2 equilibration.

Inhibitor sensitivity of pulmonary vascular carbonic anhydrase.

The inhibitor sensitivity of pulmonary vascular carbonic anhydrase (CA) was examined in situ to identify the specific isozyme responsible for vascular activity and to study its distribution in the lung. Vascular CA activity was monitored in isolated rat lungs by measuring the rate of CO2 excretion and the magnitude of postcapillary CO2-HCO(3-)-H+ disequilibria. Lungs were perfused with isotonic salines containing gluconate, sulfate, Cl-, or I-, with or without sulfonamide derivatives. Effects of a CA inhibitor purified from porcine blood plasma were also determined. Vascular CA activity was unaffected by gluconate, sulfate, Cl-, and I- (< or = 100 mM). Sulfonamides with vastly different rates of membrane permeation (i.e., readily permeating ethoxzolamide, slowly permeating acetazolamide, and membrane-impermeant quaternary ammonium sulfanilamide) were capable of accessing all vascular CA with similar rates of access. The porcine inhibitor of CA (340 nM) produced a significant, but submaximal, inhibition of vascular CA activity. The data suggest that pulmonary vascular activity reflects a high-activity membrane-bound isozyme, CA IV, which is located on the extracellular luminal surface of capillary endothelial cells.

Characteristics of the anion transport system in sea turtle erythrocytes.

Erythrocytes of Kemp's ridley sea turtle (Lepidochelys kempi) contain a 100- to 105-kDa protein that is reactive with a monoclonal antibody to the membrane domain of human erythrocyte band 3. Based on inhibition of membrane HCO(3-)-Cl- exchange with 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid (SITS), sea turtle erythrocytes were found to contain 4 x 10(6) copies of band 3 per cell. Unidirectional HCO3- transfer, specifically HCO3- out----in-Cl-in----out exchange, where subscript in----out represents transfer from inside to outside and subscript out----in represents transfer from outside to inside, was characterized by a maximal exchange rate of 1.0-1.1 nmol.cm-2.s-1, substrate affinity coefficients of 0.1-0.2 mM for HCO3- and 1.6 mM for Cl-, and an apparent inhibition constant for SITS of 0.6-1.0 microM (10 degrees C, pH 7.6). Under physiological conditions (30 degrees C, pH 7.4), the rate of net HCO3- transfer (i.e., the difference between HCO3- in----out-Cl-out----in and HCO3- out----in-Cl-in----out) was 1.13 nmol. cm-2.s-1 for cells subjected to a 5-mM decrement in CO2 content. This yields a rate coefficient for the "physiological" anion shift in sea turtle blood of 1.7 s-1, indicating that the anion shift may require 2.6 s to reach 99% completion in vivo. The erythrocyte anion shift appears to be a potential rate-limiting step for capillary CO2 exchange in these turtles.