Effects of inhibitors of enzymatic and cellular pH-regulating systems on central sympathetic chemosensitivity

Previous studies in cats using isolated NaClCO₂ perfusion of the lower brainstem demonstrated an intrinsic chemosensitivity of sympathoexcitatory bulbospinal neurones within the rostroventrolateral medulla (RVLM). In the present experiments, the effects of inhibitors of enzymatic and cellular systems, known to be involved in pH regulation, were investigated. Isolated perfusion of the lower brainstem with CO₂-enriched solutions was performed and preganglionic sympathetic nerve activity (SNA) was recorded. Drugs were locally injected into the left RVLM with glass micropipettes. Perfusion of the RVLM with CO₂enriched solutions over a period of 15 s induced a marked increase in SNA. The magnitude of absolute changes in SNA during perfusion depended on the level of basal SNA before perfusion. Microinjections of 4,4-diisothiocyanatostilbene-2,2-disulphonic acid (DIDS) and acetazolamide (ACZ) induced a marked rise in basal SNA, whereas diethylpyrocarbonate (DEPC) and ethylisopropylamiloride (EIPA) had no significant effect on basal SNA. After application of DIDS and DEPC, the peak change in SNA due to perfusion of the RVLM with CO₂-enriched solutions was slightly diminished. Furthermore, neither ACZ nor EIPA produced any significant influence on the slope, peak change and time course of the increase in SNA compared with control perfusions. We conclude that the enzymatic and cellular carrier systems tested in this study are not or only slightly involved in central sympathetic chemosensitivity.     

Does the threshold of transcutaneous partial pressure of carbon dioxide represent the respiratory compensation point or anaerobic threshold?

On reaching the respiratory compensation point (RCP) during rapidly increasing incremental exercise, the ratio of minute ventilation (V_E) to CO₂ output (VCO₂) rises, which coincides with changes of arterial partial pressure of carbon dioxide (P _aCO₂). Since P _aCO₂ changes can be monitored by transcutaneous partial pressure of carbon dioxide (PCO_2,tc) RCP may be estimated by PCO_2,tc measurement. Few available studies, however, have dealt with comparisons between PCO_2,tc threshold (T _AT) and lactic, ventilatory or gas exchange threshold (V _AT), and the results have been conflicting. This study was designed to examine whether this threshold represents RCP rather than V _AT. A group of 11 male athletes performed incremental excercise (25 W · min^–1) on a cycle ergometer. The PCO_2,tc at (44°C) was continuously measured. Gas exchange was computed breath-by-breath, and hyperaemized capillary blood for lactate concentration ([la^–]_b) and P _aCO₂ measurements was sampled each 2 min. The T _AT was determined at the deflection point of PCO_2,tc curve where PCO_2,tc began to decrease continuously. The V _AT and RCP were evaluated with VCO₂ compared with oxygen uptake (VO₂) and V_E compared with the VCO₂ method, respectively. The PCO_2,tc correlated with P _aCO₂ and end-tidal PCO₂. At T _AT, power output [P, 294 (SD 40) W], VO₂ [4.18 (SD 0.57)l · min^–1] and [la^–] [4.40 (SD 0.64) mmol · l^–1] were significantly higher than those at V _AT[P 242 (SD 26) W, VO₂ 3.56 (SD 0.53) l · min^–1 and [la^–]_b 3.52 (SD 0.75), mmol · l^–1 respectively], but close to those at RCP [P 289 (SD 37) W; VO₂ 3.97 (SD 0.43) l · min^– and [la^–]_b 4.19 (SD 0.62) mmol · l^–1, respectively]. Accordingly, linear correlation and regression analyses showed that P, VO₂ and [la^–]_b at T _AT were closer to those at RCP than at V _AT. In conclusion, the T _AT reflected the RCP rather than V _AT during rapidly increasing incremental exercise.     

Interstitial PCO2 and pH in rat hippocampal slices measured by means of a novel fast CO2/H+-sensitive microelectrode based on a PVC-gelled membrane

We describe here the construction and properties of a double-barrelled microelectrode (tip diameter 4–10 m) which permits simultaneous measurements of PCO₂ and pH, and which has a 90% response time of only one or a few seconds for a step change in PCO₂. The fast response of the CO₂-sensitive barrel is due to (i) the use of a PVC-gelled (tridodecylamine-containing) membrane solution which enables the construction of extremely short ( 4 m), yet mechanically stable, membrane columns, and (ii) the presence of carbonic anhydrase in the filling solution. Recordings made in the pyramidal layer of area CA1 in rat hippocampal slices showed that the deviation in the acid direction of the basal interstitial pH (pH₀) from that of the perfusion solution was attributable to a higher PCO₂ level within the tissue. Most of the late acid shift evoked by stimulation of the Schaffer collaterals (5- to 20-s trains at 10 Hz) could also be explained on the basis of an accumulation of interstitial CO₂ at a constant HCO ₃ ^– concentration. This conclusion was supported by the finding that inhibition of extracellular carbonic anhydrase activity by 10 M benzolamide completely abolished the activity-induced fall in pH₀, but not the increase in PCO₂. The initial stimulus-induced alkalosis was accompanied by a slight decrease in PCO₂ only, implying a parallel increase in the interstitial HCO ₃ ^– concentration. Benzolamide produced a dramatic enhancement of the early alkaline shift as well as of the simultaneous fall in PCO₂. The latter effect of the drug unmasks a cellular CO₂ sink that is induced by neuronal activity.     

Effect of H+ and elevated PCO 2on membrane electrical properties of rat cerebral arteries

Some membrane electrical properties of muscle cells from the middle cerebral artery of the rat were recorded with intracellular microelectrodes. The resting membrane potential (E _m) of this preparation was –63 mV. Reduction of extracellular pH to 7.0 in the face of a constantP _{CO 2}of 40 mm Hg had no significant effect onE _m. Similarly the slope of the steady-state voltage/current curves was not different at pH 7.0 compared to control at pH 7.4. In marked contrast, whenP _{CO 2}was elevated to around 60 to 70 mm Hg there was a rapid hyperpolarization and reduction in the slope of the voltage current curve suggesting an increased conductance for one or more ionic species. In addition elevation ofP _{CO 2}increased the slope of theE _m vs. log[K]₀ curve from 46 mV/decade to 59 m V/decade which is in good agreement with a Nernstian potential for a K⁺ selective membrane. These data suggest that while the smooth muscle cells of rat cerebral arteries are relatively insensitive to a small reduction in extracellular pH; reduction of intracellular pH by elevatingP _{CO 2}induces hyperpolarization by increasing K⁺ conductance (g _k). However, it is not clear from these experiments if theP _{CO 2}effects are mediated entirely by changes in pH or if there is a direct membrane action of CO₂.This work is supported by Grant no. HL27862     

Comparison of arterial,end-tidal and transcutaneous PCO2 during moderate exercise and external C02 loading in humans

Summary Static relationships between arterial, transcutaneous[/p] and end-tidal PCO₂ (P _aCO₂, P _tc CO ₂, P _etCO₂) as well as the dynamic relationship between P _etCO₂ and P _tcCO₂ were studied during moderate bicycle ergometer exercise with and without external C0₂ loading. The exercise pattern consisted of 5-min intervals of constant power at 40 W and 100 W and 900 s of randomised changes between these two power levels. The external CO₂ loading was achieved by means of controlled variations of inspiratory gas compositions aimed at a constant P _etCO₂ of 6.5 kPa (49 mm Hg). The P_etO2 was regulated at 17.3 kPa (130 mm Hg). Under steady-state conditions all PCO₂ parameters showed close linear relationships. P _aCO₂/P _tcCO₂ was near to identity while the P _etCO₂ systematically overestimated changes in P _aCO₂. No relationship showed a significant influence of the exercise intensity. Transients of P _tcCO₂ are considerably slower than P _etCO₂ transients. The dynamic relationship between both parameters was found to be independent of whether internal or external C0₂ loadings were applied. It is concluded that the combination of P _etCO₂ and P _tcCO₂ measurements allows an improved non-invasive assessment of P _aCO₂. While P _etC0₂ better reflects the transients, P _tcCO₂ can be employed to determine slow changes of the absolute P _aCO₂.     

APCO2 surface electrode working on the principle of electrical conductivity

APCO₂ electrode working on the principle of electrical conductivity is described. The calibration curve can be linearized according to the formula . This linearity has been tested in thePCO₂ range of 0.93–9.33 kPa (7–70 Torr). For the experiments electrodes are used which have conductivity values of about 50 nS and drifts of maximally 5%/h at aPCO₂ of 5.33 kPa (40 Torr). The response time (T ₉₀) is about 20 s. The temperature sensitivity is 2.4 nS/1 K between 298K–310K. The standard error of the measurements is =0.33 nS. With these electrodes tissuePCO₂ can be measured on the surface of various organs.     

Ventilatory response to oscillating and non-oscillatingPa CO 2 in the anesthetized cat

Summary In 11 adult cats, lightly anesthetized with chloralose-urethane, blood from both common carotid arteries was led into a plastic chamber of 15–20 ml and returned to the carotids at a point 1.5 cm more cranial. By doing so arterial blood was assumed to pool within the chamber and lose itsP _{CO 2} oscillations which are normally known to exist as a result of the respiratory cycle. In control periods blood bypassed the chamber, thus maintaining respiratoryP _{CO 2} oscillations. Spontaneous ventilation was measured spirometrically. The animals were breathing pure O₂.Results. 1. When the sinus (carotid) nerves were intact or sectioned there was no significant difference in ventilation before or after switching from non-oscillating to oscillatingPa _{CO 2}. 2. When the vertebral arteries were ligated a drop in ventilation occurred after turning to oscillatingPa _{CO 2} which was followed by a slight rise above control values after 30–50 sec. This phenomenon was independent of sinus nerve integrity. Thus in hyperoxie condition the smallPa _{CO 2} oscillations known to occur in phase with respiration do not seem to provide a respiratory stimulus to resting ventilation above that generated by the mean level ofPa _{CO 2}. The ventilatory depression after vertebral artery ligation must at this time remain unexplained.     

Der Einfluß desP A CO 2 auf die Wirkung der Sinusnervenreizung bei intakten und ausgeschalteten Nn. vagi

Summary In rabbits rebreathing oxygen one carotid sinus nerve was stimulated repeatedly by electrical stimuli of constant intensity, both before and after inactivation of vagi by cold blocking or sectioning. In animals with intact vagi the reflex hyperpnea elicited by the nerve stimulation decreased only slightly with increasing hypercapnia. After inactivation of the vagi the stimulatory effect was, at normalP _{A CO 2}, mostly greater than in the intact condition. But with increasing hypercapnia the decrease of the reflex hyperpnea was generally steeper than in the intact preparation, sometimes reaching even lower values than before inactivation. The depressant effects of the stimulation on systemic blood pressure and heart rate were not distinctly changed by the increase ofP _{A CO 2}, neither with intact nor with inactivated vagi. It is concluded that the ventilatory effect of chemoreceptor afferents is modified both by centralP _{CO 2} and by vagal afferents.

Mit Unterstützung der Deutschen Forschungsgemeinschaft (Wi 165).     

CO2 responsivity in the mouse measured by rebreathing

We have modified the rebreathing method to study CO₂ responsivity in very small mammals. Tidal volume (V _T) and frequency (f) of pentobarbital-anesthetized mice were measured during rebreathing from a closed circuit, primed with 95% O₂, 5% CO₂, through which the gas was constantly circulated at 0.5 l·min^–1. The circuit consisted of T-tube from a plethysmograph, Tygon tubing with compliant element, CO₂ analyzer and pump, in series. CircuitPCO₂ (PctCO₂), which was recorded continuously during spontaneous breathing, rapidly equilibrated with end-tidalPCO₂. CO₂ response curves were constructed from extrapolated minute ventilation ( ),V _T,f and parameters of breath-to-breath timing, respectively, onPctCO₂. Analyses of slopes of the response curves, change from onset of rebreathing to peak response, andPctCO₂ at which the response peaked revealed that CO₂ stimulates by increasingf andV _T and that this is effected by facilitation of central inspiratory-expiratory phase switching and inspiratory drive mechanisms. However, the stimulatory effect of CO₂ on phase switching was not sustained, with maximal effect occurring before peak . The advantages and facility of the modified rebreathing method make it suitable for studies of other small mammals, including neonates.     

The involvement of expiratory termination in the vagally mediated facilitation of ventilatory CO2 responsiveness during hyperoxia

In anaesthetized rabbits the influence of differential vagal cold blockade on the ventilatory response to inhaled CO₂ during hyperoxia was investigated.Following total inactivation, the relationship between ventilation ( ) and arterialPCO₂ (P _aCO₂) was shifted to the left and steepened slightly over a range of modest hypercapnia, but was progressively flattened as hypercapnia intensified. The latter effect, suggestive of a vagally mediated facilitation of ventilatory CO₂ responsiveness, was studied further.Differential vagal cold blockade to a temperature (5–11°C) which abolished the Breuer-Hering inflation reflex (end-inspiratory tracheal occlusion no longer eliciting a prolongation of expiratory duration,T _E) had no effect on either during normocapnia or at a substantial level of hypercapnia. Only with further vagal cooling to 0°C did the ventilatory depression during hypercapnia emerge, largely becauseT _E failed to shorten in response to the hypercapnic stimulus.It is concluded that the integrity of expiratory-terminating mechanisms is crucial for the manifestation of the vagally mediated facilitation of and its CO₂ responsiveness which is evident during hyperoxic hypercapnia. A possible role is suggested for lung epithelial irritant receptors or for the tonic late-expiratory activity from pulmonary stretch receptors.Supported by the Deutsche Forschungsgemeinschaft, SFB 114Preliminary reports of this work have been presented in Pflügers Arch 355: (Suppl) R47 (1975); 377: (Suppl) R54 (1978) and in Proc. XXVIII. Int. Congr. of Physiol. Sciences, Budapest, Vol VIV, 515 (1980)     

The role of fitness on VO2 and VCO2 kinetics in response to proportional step increases in work rate

Summary The purpose of this study was to determine the effect of fitness and work level on the O₂ uptake and CO₂ output kinetics when the increase in work rate step is adjusted to the subject's maximum work capacity. Nine normal male subjects performed progressive incremental cycle ergometer exercise tests in 3-min steps to their maximum tolerance. The work rate step size was selected so that the symptom-limited maximum work rate would be reached in four steps at 12 min in all subjects. Oxygen consumption (VCO₂) and carbon dioxide production VCO₂ were calculated breath by breath. For the group, the time (mean, SEM) to reach 75% of the 3-min response (T _0.75) for VO₂ increased significantly (P<0.01) at progressively higher work rate steps, being 53.3 (5.5) s, 63.5 (4.6) s, 79.5 (5.0) s, and 94.5 (5.8) s, respectively. In contrast, T _0.75 for VCO₂ did not change significantly [74.9 (7.4) s,. 75.6 (5.0) s, 85.1 (5.3) s, and 89.4 (6.3) s, respectively]. VCO₂ kinetics were slower than VO₂ kinetics at the low fractions of the subjects' work capacities but were the same of faster at the high fractions because of the slowing of VO₂ kinetics. The first step showed the fastest rise in VO₂. While VO₂ kinetics slowed at each step, they were faster at each fraction of the work capacity in the fitter subjects. The step pattern in VO₂ disappeared at high work rates for the less fit subjects. The heart rate response paralleled that of VO₂. We conclude that VO₂ and VCO₂ kinetics are slower in the less fit subjects but only VO₂ kinetics are significantly attenuated in response to proportional step increases in work rate.     

Cellular mechanism of force development in cat middle cerebral artery by reducedPCO2

These studies were undertaken to determine the effect of reducing aPCO₂ below physiological levels on cat middle cerebral artery. Upon reduction ofPCO₂ from 37 to 14 torr (pH 7.4) we observed membrane depolarization and force development. ReducingPCO₂ decreased the slope of theE _m vs. log [K]_o curve and increased the slope of the steady-state I/V relationship suggesting that the change inE _m was due to reduction of outward K⁺ conductance (g _k). Elevation of pH from 7.37 to 7.6 had a very similar effect on these cerebral arterial muscle cells, depolarizing the muscle membrane (reducing theE _m vs. log [K]_o curve) and increasing the slope of the I/V relationship to statistically equivalent values as reduction ofPCO₂. ReturningPCO₂ from 14 to 37 torr rapidly relaxed these preparations, but only transiently. This relaxation was followed by a rebound contraction within 3 min, demonstrating a transient nature for the action of elevatingPCO₂ in cerebral arteries. The response to changing pH_o followed a slower time course but did not change with time. These studies demonstrate that both elevated pH_o and reducedPCO₂ activate cerebral arterial muscle by a mechanism which includes reduction ing _k. However, it can not be determined if these similar responses and reduction, ofg _k are mediated by changing pH_i or mediated through different mechanisms. It is possible that pH_o andPCO₂ can modify cerebral arterial tone by direct mechanisms and not necesarily by their effect on pH_i. It is clear, however, that reduction ofPCO₂ and elevation of pH_o both activate cerebral arterial muscle by a mechanism which includes reduction ofg _k.This study was supported by NIH grant no. HL-32871. Dr. Harder is an established investigator of the American Heart Association     

Transmural gradient of tissue gas tensions in the canine left ventricular myocardium during coronary clamping and reactive hyperemia

Mass spectrometry was used for the continuous, simultaneous and quantitative measurement of oxygen (PO₂) and carbon dioxide (PCO₂) partial pressures in the subendocardial and subepicardial layers of the left ventricle in 11 anaesthetized ventilated dogs. Under control conditions,PO₂ was significantly lower in the subendocardium (13.5±4.5 mm Hg) than in the subepicardium (20.7±2.3 mm Hg), whereasPCO₂ did not differ significantly (43±8.8 and 51±9.2 mm Hg respectively). These variables were not correlated with blood pressure or coronary blood flow. Subendocardial and subepicardialPO₂ decreased less than 5 s after coronary occlusion. These changes were more rapid and severe in the subendocardium. After occlusion for 90 s: subendocardialPO₂ was 4.1±6.3 mm Hg while subepicardialPO₂ was 6.7±15.0 mm Hg (P<0.05).PCO₂ reached peak values of 56±25 mm Hg subendocardial and 82±22 mm Hg subepicardial at 2.67±0.71 min and 3.43±0.93 min after coronary clamping. A reactive hyperemia occurred after coronary unclamping with different time courses and amplitudes for systolic and diastolic stroke flows whilePO₂ recovered with different kinetics. SubendocardialPO₂ increased with a lower initial slope, probably in relation with the delay in the diastolic hyperemia. The observed delayed subendocardial hyperoxia, unrelated to the hyperemia, may indicate a delay in the recovery of normal work and metabolism in the inner layers of the myocardium.     

The effects of CO2 and pH on the spontaneous activity of the taenia coli of guinea-pig

Summary Mechanical and electrical activity of the isolated guinea-pig taenia coli were recorded in bicarbonate-, Tris- or phosphate-buffered test solutions. In normal solution (pCO₂ 5%, pH 7.4), typical minute-rhythmical fluctuations of activity occurred, whereby activity was present for approximately 40% of the total time (active time), the other 60% being activity-free intervals. At constant extracellular pH, low CO₂ content increased active time, high CO₂ content reduced it. Extracellular alkalinization at constantpCO₂ also diminished active time, even in CO₂-free medium, whereas acidification raised it, sometimes causing continuous activity. At constant bicarbonate-buffer, changes of CO₂ content, i.e. accompanied by corresponding changes in pH, affected the active time much less than did CO₂ alterations in an isohydric medium. The test solutions had no major effect on frequency of synchronized spike discharges, in contrast to their actions on the minuterhythmical activity. Since CO₂ can pass through the cell membrane more easily than ions regulating intra- and extracellular pH, the observed effects are best explained by changes in the transmembrane pH-gradient. A drop in active time would be due to a relative intracellular acidification; continuous activity, on the other hand, due to an opposite change in transmembrane pH-gradient, i.e. a relative intracellular alkalinization.This work was supported by the Deutsche Forschungsgemeinschaft     

Analysis of alveolar PCO 2 control during the menstrual cycle

We attempted to analyze how is regulated during progesterone-induced hyperventilation in the luteal phase. A model for the CO₂ control loop was constructed, in which the function of the CO₂ exchange system was described as and that of the CO₂ sensing system as . Using this model, we estimated (1) the primary increase in produced by progesterone stimulation and (2) the effectiveness (E) of the loop to regulateP _{A CO 2}, defined as P _{A CO 2} (op)/P _{A CO 2} (cl) in which op signifies open-loop and cl, closed-loop. These respiratory variables were investigated throughout the menstrual cycle in 8 healthy women. During the luteal phase, on average, increased by 9.4% andP _{A CO 2},B andH decreased by 0.33 kPa (2.5 mm Hg), 0.47 kPa (3.5 mm Hg) and 13.6%, respectively, whileS and did not change significantly. (op) increased progressively on successive days of the luteal phase whileE remained unchanged at a value of 7.9, thus there was a progressive decrease inP _{A CO 2}. The decrease inH was considered to lessen P _{A CO 2} (op) and so reduce the final deviation ofP _{A CO 2} (P _{A CO 2} (cl)) during the luteal phase. The decrease inB was found to be dependent on (op).     

Ventilatory response of prepubertal boys and adults to carbon dioxide at rest and during exercise

Summary The aim of this study was to determine whether the greater ventilation in children at rest and during exercise is related to a greater CO₂ ventilatory response. The CO₂ ventilatory response was measured in nine prepubertal boys [10.3 years (SD 0.1)] and in 10 adults [24.9 years (SD 0.8)] at rest and during moderate exercise ( CO₂ = 20 ml·kg^–1·min^–1) using the CO₂-rebreathing method. Three criteria were measured in all subjects to assess the ventilatory response to CO₂: the CO₂ sensitivity threshold (Th), which was defined as the value of end titalPCO₂ (P _ETCO₂) where the ventilation increased above its steady-state level; the reactivity slope expressed per unit of body mass (SBM), which was the slope of the linear relation between minute ventilation ( _E) andP _ETCO₂ above Th; and the slope of the relationship between the quotient of tidal volume (V _T) and inspiration time (t _I) andP _ETCO₂ (V _T ·t _I ^–1 ·P _ETCO₂ ^–1) values above Th. The _E,V _T, breathing frequency (f _R), oxygen uptake ( O₂), and CO₂ production ( CO₂) were also measured before the CO₂-rebreathing test. The following results were obtained. First, children had greater ventilation per unit body weight than adults at rest (P<0.001) and during exercise (P<0.01). Second, at rest, onlyV _T ·t _I ^–1 ·P _ETCO₂ ^–1 was greater in children than in adults (P<0.001). Third, during exercise, children had a higher S_BM (P < 0.02) andV _T ·t _I ^–1 ·P _ETCO₂ ^–1 (P<0.001) while Th was lower (P<0.02). Finally, no correlation was found between _E/ CO₂ and Th while a significant correlation existed between _E/ CO₂ and S_BM (adults,r=0.79,P<0.01; children,r=0.73,P<0.05). We conclude that children have, mainly during exercise, a greater sensitivity of the respiratory centres than adult. This greater CO₂ sensitivity could partly explain their higher ventilation during exercise, though greater CO₂ production probably plays a role at rest.     

Physiological responses to intermittent and continuous exercise at the same relative intensity in older men

We investigated the physiological responses in older men to continuous (CEx) and intermittent (IEx) exercise. Nine men [70.4 (1.2) years, O_2peak: 2.21 (0.20) l min^–1; mean (SE)] completed eight exercise tests (two CEx and six IEx) on an electronically braked cycle ergometer in random order. CEx and IEx were performed at 50% and 70% O_2peak. IEx was performed using 60sE:60sR, 30sE:30sR and 15sE:15sR exercise to rest ratios. The duration of exercise was adjusted so that the total amount of work completed was the same for each exercise test. Oxygen uptake (O₂), minute ventilation (_E) and heart rate (HR) were measured at the mid-point of each exercise test. Arterialised blood samples were obtained at rest and during exercise and analysed for pH and PCO₂. At the same relative intensity (50% or 70% O_2peak), IEx resulted in a significantly lower (P<0.01) O₂, _E and HR than CEx. There were no significant differences (P>0.05) in O₂, _E and HR measured at the mid point of the three exercise to rest ratios at 50% and 70% O_2peak. pH and PCO₂ during CEx and IEx at 50% O_2peak were not significantly different from rest. CEx performed at 70% O_2peak resulted in significant decreases (P<0.05) in pH and PCO₂. There was a significant decrease (P<0.05) in pH only during the 60sE:60sR IEx at 70% O_2peak. Changes in arterialised PCO₂ during the 60sE:60sR, 30sE:30sR and 15sE:15sR at both 50% and 70% O_2peak exercise tests were not significant. When exercising at the same percentage of O_2peak and with the total amount of work fixed, IEx results in significantly lower physiological responses than CEx in older men. All results are given as mean (SE).     

Fixed acid and carbon dioxide bohr effects as functions of hemoglobin-oxygen saturation and erythrocyte pH in the blood of the frog,Rana temporaria

Using a thin film, dynamic recording technique, the pH sensitivity of the oxygen equilibrium (Bohr effect) of whole blood in the frogRana temporaria, and its dependence on CO₂ and fixed acids and on plasma and erythrocyte pH values were measured. Under standard conditions (20°C,P _{CO 2}=14.7 mm Hg, pH=7.65) the oxygen equilibrium could be described by a P₅₀ value of 38 mm Hg andn ₅₀ of 1.8. Hill plots of the oxygen equilibria showed increased cooperativity in oxygen binding with increasing saturation (n ₂₀ 1.2,n ₈₀ 4.0). Values of the fixed acid and CO₂ Bohr factors ({ie7-1} and {ie7-2}, respectively) were similar at specific saturations (S_{20, 50, 80}) but showed saturation dependence with high values occurring at high saturation. The same statements also hold for the intracellular Bohr factors (derived from the relation between blood P₅₀ and erythrocyte pH) although the values of both {ie7-3} and {ie7-4} now were greater than those related to blood pH.     

Hyperpnoea and CO2 sensitivity of the respiratory centres during exercise

Summary The aim of this study was to specify whether exercise hyperpnoea was related to the CO₂ sensitivity of the respiratory centres measured during steady-state exercise of mild intensity. Thus, ventilation , breathing pattern [tidal volume (V _T), respiratory frequency (f), inspiratory time (T _I), total time of the respiratory cycle (T _TOT),V _T/T _I,T _I/T _TOT] and CO₂ sensitivity of the respiratory centres determined by the rebreathing method were measured at rest (SCO_{2 re}) and during steady-state exercise (SCO_{2 ex}) of mild intensity [CO₂ output =20 ml·kg⁻¹·min⁻¹] in 11 sedentary male subjects (aged 20–34 years). The results showed that SCO_{2 re} and SCO_{2 ex} were not significantly different. During exercise, there was no correlation between and SCO_{2 ex} and, for the same , all subjects had very close values normalized for body mass (bm), regardless of their SCO_{2 ex} ( =1.44 l·min⁻¹·kg⁻¹ SD 0.10). A highly significant positive correlation between SCO_{2 ex} andV _T (normalised for bm) (r=0.80,P<0.01),T _I (r=0.77,P<0.01) andT _TOT (r=0.77,P<0.01) existed, as well as a highly significant negative correlation between SCO_{2 ex} and (normalised for bm^−0.25) (r=−0.73,P<0.01). We conclude that the hyperpnoea during steady-state exercise of mild intensity is not related to the SCO_{2 ex}. The relationship between breathing pattern and SCO_{2 ex} suggests that the breathing pattern could influence the determination of the SCO_{2 ex}. This finding needs further investigation.     

Correlation of discharges of rostral ventrolateral medullary neurons with the low-frequency sympathetic rhythm in rats

The rostral ventrolateral medulla (RVLM) is critically important in the generation of sympathetic activity. The purpose of this study was to investigate whether discharges of RVLM neurons contribute to low-frequency (LF) sympathetic rhythms. Blood pressure (BP), renal sympathetic nerve activity (SNA), and neuronal activity in the RVLM were simultaneously recorded in seven anesthetized, paralyzed, and artificially ventilated rats. Fifty-one RVLM neurons were recorded and classified into three differential functional groups according to their activities related to baroreceptor input. Those in the category of spike firing inhibited by a BP increase (BP^I) and which excited sympathetic discharges was the most abundant (24%). Coherence analysis was used to examine the relationship of the firing frequency of RVLM neurons with the LF (0.2–0.8 Hz) rhythm of SNA. Forty-one percent of RVLM neurons showed a significant correlation to LF rhythms, and BP^I neurons with sympathoexcitatory properties were the major contributors. In another 4 baroreceptor-denervated rats, 36 RVLM neurons were recorded. In these rats, RVLM neuronal activities no longer changed with BP fluctuations. Nevertheless, more than 40% of RVLM neurons were sympathoexcitatory, and 36% of RVLM neurons were still correlated with the LF SNA rhythm. Our results suggest that there are RVLM neurons involved in generating the LF rhythm in SNA and that the baroreflex can induce the participation of more neurons in LF rhythm generation.