Cardioventilatory effects of acclimatization to aquatic hypoxia in channel catfish

The mechanisms responsible for altering cardioventilatory control in vertebrates in response to chronic hypoxia are not well understood but appear to be mediated through the oxygen-sensitive chemoreceptor pathway. Little is known about the effects of chronic hypoxia on cardioventilatory control in vertebrates other than mammals. The purpose of this study was to determine how cardioventilatory control and the pattern of response is altered in channel catfish (Ictalurus punctatus) by 1 week of moderate hypoxia. Fish were acclimatized for 7 days in either normoxia (P(O(2)) approximately 150 Torr) or hypoxia (P(O(2)) approximately 75 Torr). After acclimatization, cardioventilatory, blood-gas and acid/base variables were measured during normoxia (P(O(2)) 148+/-1 Torr) then at two levels of acute (5 min) hypoxia, (P(O(2)) 72.6+/-1 and 50.4+/-0.4 Torr). Ventilation was significantly greater in hypoxic acclimatized fish as was the ventilatory sensitivity to hypoxia (Delta ventilation/Delta P(O(2))). The increase in ventilation and hypoxic sensitivity was due to increases in opercular pressure amplitude, gill ventilation frequency did not change. Heart rate was greater in hypoxic acclimatized fish but decreased in both acclimatization groups in response to acute hypoxia. Heart rate sensitivity to hypoxia (Delta heart rate/Delta P(O(2))) was not affected by hypoxic acclimatization. The ventilatory effects of hypoxic acclimatization can be explained by increased sensitivity to oxygen but the effects on heart rate cannot.     

Conductance of the sodium channel in myelinated nerve fibres with modified sodium inactivation.

1. Na current fluctuations in nodes of Ranvier were measured under voltage clamp conditions as described in the preceding paper (Conti, Hille, Neumcke, Nonner & St?mpfli, 1976) and analysed in terms of power spectral density calculated for frequencies between 30 Hz and 5 kHz. 2. External (10(-5) g/ml.) Leiurus scorpion venom or Anemonia Toxin II (3 X 10(-5) g/ml.) or internal 20 mM iodate were applied in order to remove or slow down inactivation in part of the Na channels. The treatment increased the steady-state Na current during the noise measurement one-to eight fold over that in normal fibres. 3. Noise spectra were interpreted as the sum of 1/f noise and noise SNa(f) due to all-or-none, open-close transitions of single Na channels. The drug effects on the inactivation could be accounted for either by assuming two populations of channels, one with and one without inactivation, or by postulating a single population with modified inactivation characteristics. 4. Except for an increase in amplitude, the fluctuation spectra SNa(f) were similar to the ones in normal nodes. Again, the time constants taum obtained from the fit of the spectra agreed within a factor of 2 with the values of taum found in the macroscopic Na currents. 5. From the fluctuation spectra, single Na channel conductances gamma of 5-4 +/- 0-4 pS (iodate), 6-7 +/- 0-5 pS (Leiurus) and 7-0 +/- 0-6 pS (Anemonia) were calculated. The value of gamma was not significantly voltage dependent. 6. Our observations indicate that inactivation of Na channels can be modified with at most small effects on the microscopic properties of the activation process and on the conductance of the open channel. They suggest that the h mechanism normally produces all-or-none, open-close changes of conductance.     

Cerebral blood flow and oxygen consumption in the newborn dog

Cerebral blood flow (CBF), CBF responses to changes in arterial CO2 tension, and cerebral metabolic rate for oxygen (CMRO2) were measured in newborn dogs, by means of a modification of the Kety and Schmidt technique employing 133Xe. Mongrel dogs of 1-7 days of age were paralyzed and passively ventilated with 70% N2O and 30% O2. CBF was derived by analysis of paired serial 20-microliter samples of arterial and of cerebral venous blood from the superior sagittal sinus. At an arterial PCO2 of 36.9 +/- 3.7 Torr and a mean arterial blood pressure of 62 +/- 10 Torr, CBF was 23 +/- 8 ml/min per 100 g. The arteriovenous oxygen content difference averaged 5.6 vol%, and CMRO2 was 1.13 +/- 0.30 ml O2/min per 100 g. CBF increased or decreased by 0.58 ml/min/100 g per Torr change in PCO2. Our results suggest that in the newborn, basal CBF and CBF responses to CO2 are considerably lower than in the adult and parallel the lower metabolic needs of the newborn brain.     

Effect of intermittent hypoxia on cardiovascular function,adrenoceptors and muscarinic receptors in Wistar rats

The usual model of intermittent hypoxia (sleep apnoea) corresponds to repeated episodes of hypoxia from a few seconds to a few hours interspersed with episodes of normoxia. The aim of this study was to evaluate in rats the effect of two periods of intermittent exposure for 2 months to hypoxia (IHX1, 24 h in hypoxia (428 Torr), 24 h in normoxia; IHX2, 48 h in hypoxia (428 Torr), 24 h in normoxia) as a new model of hypoxia simulating intermittent exposure to high altitude experienced by Andean miners. We assessed the haematological parameters, time course of resting heart rate and systolic blood pressure. We also evaluated the expression of adrenergic and muscarinic receptors. IHX1 and IHX2 produced an increase in haematocrit, haemoglobin concentration and mean corpuscular volume as previously seen in most hypoxic models. IHX1 and IHX2 induced a similar sustained elevation of systolic blood pressure (132 +/- 2 and 135 +/- 3 mmHg, respectively, vs. the control level of 121 +/- 16 mmHg) after 10 days of exposure without change in heart rate. Right ventricular (RV) hypertrophy (225 +/- 13 and 268 +/- 15 mg g(-1), vs. 178 +/- 7 mg g(-1) and downregulation of alpha1-adrenoceptor (RV: 127 +/- 21 and 94 +/- 16 fmol mg(-1) vs. 157 +/- 8 fmol mg(-1); left ventricle (LV): 141 +/- 5 and 126 +/- 9 fmol mg(-1) vs. 152 +/- 5 fmol mg(-1)) have been found in both groups, with right ventricular hypertrophy being greater and alpha1-adrenoceptor density being lower in IHX2 than in HX1 groups. These data indicate that both parameters are related to the time of exposure to hypoxia. IHX1 and IHX2 produced the same magnitude of upregulation of muscarinic receptors (LV, 60%; RV, 40%), and no change in beta-adrenoceptors. In conclusion, exposure to intermittent hypoxia led to polycythaemia and RV hypertrophy as observed in other types of hypoxia. A specific cardiovascular response was seen, that is an increase in blood pressure without change in heart rate, which was different from the one observed in episodic and chronic hypoxia. Furthermore, this model involved specific modifications of alpha1-adrenergic and muscarinic expression.     

Characterization of potassium channels in respiratory cells

Potassium channels present in the basolateral membrane of respiratory epithelial cells play an important role in the process of chloride secretion. Utilizing the patch clamp technique, we examined human cultured respiratory epithelial cells derived from patients with cystic fibrosis (CF) and normals individual (N) for the existence of and for the properties of K+ channels. We obtained qualitatively and quantitatively identical results for both preparations (CF and N). K+ channels were spontaneously present in cell attached patches. The channels showed burst appearance with rapid flickering within the bursts. When the pipette was filled with 145 mmol/l KCl, a mean conductance of 131 +/- 25 pS (n = 15) was read from the I/V-curve at a clamp voltage (Vc) of 0 mV. After excision, the conductance read from the I/V-curve at Vc = 0 mV was 212 +/- 11 pS (Pipette: 145 mmol/l KCl, bath: 145 mmol/l NaCl) (n = 61). With NaCl in the pipette and KCl in the bath, a similar conductance was obtained (g = 210 pS; n = 2). When both, pipette and bath contained KCl, the conductance was increased to 302 +/- 19 (n = 7). The channel was highly selective for potassium over sodium: PK + /PNa + greater than 40. The channel open probability was only slightly voltage dependent i.e. the open probability increased slightly with depolarisation. For most of the channels one open time constant (to = 6.3 +/- 1.6 ms; n = 22) and one closed time constant (tc = 1.8 +/- 0.3 ms; n = 21) was obtained.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of decreased oxygen availability on NADH and lactate contents in human skeletal muscle during exercise

Eight men cycled for 5 min at 120 +/- 6 W (mean +/- SE) at which O2 uptake was 50% of its maximal normoxic value, breathing room air (21% O2; normoxia) on one occasion and 11% O2 in N2 (respiratory hypoxia/hypoxic--Resp. Hx.) on the other. Biopsies were taken from the quadriceps femoris muscle. Oxygen uptake during exercise was not significantly different between Resp. Hx (1.59 +/- 0.08 1 min-1) and normoxia (1.55 +/- 0.08 1 min-1). At rest, muscle lactate was the same under both conditions but was four times higher after Resp. Hx (33.2 +/- 5.2 mmol kg-1 dry wt) than normoxic cycling (8.6 +/- 1.0 mmol kg-1 dry wt; P less than 0.01). The muscle lactate/pyruvate (which is proportional to cytosolic NADH/NAD) was significantly higher after Resp. Hx.(76 +/- 19) than after normoxic cycling (26 +/- 2; P less than 0.05). At rest, analytically determined NADH averaged 0.14 +/- 0.02 mmol kg-1 dry wt under both conditions. However, exercise during Resp. Hx. resulted in a significantly higher NADH content (0.17 +/- 0.01) than exercise during normoxia (0.12 +/- 0.01; P less than 0.01). Indirect evidence indicates that the difference in muscle NADH reflects a difference in the mitochondrial redox state (Sahlin & Katz 1986). The increased muscle NADH during Resp. Hx. therefore indicates a relative lack of O2 at the cellular level (muscle hypoxia). It is suggested that the increased lactate production during Resp. Hx. is a consequence of the cellular adaptation to muscle hypoxia (i.e. increases in cytosolic ADP, AMP, Pi and NADH).     

The effect of hypoxia on pulmonary O2 uptake, leg blood flow and muscle deoxygenation during single-leg knee-extension exercise

The effect of hypoxic breathing on pulmonary O(2) uptake (VO(2p)), leg blood flow (LBF) and O(2) delivery and deoxygenation of the vastus lateralis muscle was examined during constant-load single-leg knee-extension exercise. Seven subjects (24 +/- 4 years; mean +/-s.d.) performed two transitions from unloaded to moderate-intensity exercise (21 W) under normoxic and hypoxic (P(ET)O(2)= 60 mmHg) conditions. Breath-by-breath VO(2p) and beat-by-beat femoral artery mean blood velocity (MBV) were measured by mass spectrometer and volume turbine and Doppler ultrasound (VingMed, CFM 750), respectively. Deoxy-(HHb), oxy-, and total haemoglobin/myoglobin were measured continuously by near-infrared spectroscopy (NIRS; Hamamatsu NIRO-300). VO(2p) data were filtered and averaged to 5 s bins at 20, 40, 60, 120, 180 and 300 s. MBV data were filtered and averaged to 2 s bins (1 contraction cycle). LBF was calculated for each contraction cycle and averaged to 5 s bins at 20, 40, 60, 120, 180 and 300 s. VO(2p) was significantly lower in hypoxia throughout the period of 20, 40, 60 and 120 s of the exercise on-transient. LBF (l min(-1)) was approximately 35% higher (P > 0.05) in hypoxia during the on-transient and steady-state of KE exercise, resulting in a similar leg O(2) delivery in hypoxia and normoxia. Local muscle deoxygenation (HHb) was similar in hypoxia and normoxia. These results suggest that factors other than O(2) delivery, possibly the diffusion of O(2,) were responsible for the lower O(2) uptake during the exercise on-transient in hypoxia.     

Activation of Ionic Channels by Deoxycholate in Frog and Human Cell Lines

Humans, after extensive ileal resection, frequently suffer from diarrhoea, which may be due to an increased delivery of deoxycholate (DOC) to the large intestine. In the frog skin the addition of DOC (0.5 mM) to the apical side induced the activation of amiloride-sensitive Na+ channels and an increase in the unidirectional Cl- fluxes. Here we used two established cell lines (A6 and Caco2) to study the effect of DOC on ion channels at cell and membrane level using the patch-clamp technique. In A6 cells subcultured directly on Petri dishes and studied in the whole-cell configuration, DOC induced an increase in cell conductance of 110.3 +/- 4 pS pF-1 (N = 8) which was reduced to 89 +/- 14 pS pF-1 (N = 8) by the addition of DIDS (0.5 mM), The absolute values of these two effects were not statistically different (P < 0.2). In Caco2 cells, the addition of DOC (0.5 mM) induced, after 1 min, an increase in cell conductance of 583 +/- 16 pS pF-1 (N = 8) which was reduced to 560.4 +/- 16 pS pF-1 (N = 8) by DIDS (0.5 mM) and N-phenylanthranilic acid (DPC; 0.5 mM). The two values were not statistically different (P < 0.4). In Caco2 cells subcultured under the same conditions, DOC induced an increase in cell conductance of 1710 +/- 64 pS pF-1 (N = 6). Subsequent addition of amiloride (0.1 mM) reduced the cell conductance to 1558 +/- 33 pS pF-1 (N = 6). These two mean values were statistically different allowing for an error of the second kind < 0. 05. In cells in which DOC produced a conductance increase of 1010 +/- 10 pS pF-1, gadolinium (0.5 mM) induced a fall in cell conductance of 1800 +/- 10 pS pF-1. In Caco2 cells, addition of DOC (0.5 mM) to the bath reversibly induced the appearance of or an increase in channel activity in patches studied in cell-attached and excised inside-out configuration. In inside-out experiments (N = 13) DOC (0. 5 mM) induced the appearance of channel activity with conductances and reversal potentials (Er) of 27.7 +/- 1.9 pS and 0.8 +/- 5.7 mV, respectively. In cell-attached patches (N = 13) these values were 24.9 +/- 4.4 pS and -18.1 +/- 6.4 mV. In excised inside-out patches from Caco2 cells, subjected to electrochemical gradients for Na+, K+ and Cl-, (+85, -85 and 0 mV, respectively), addition of DOC also induced an increase in the baseline conductance and a shift in the reversal potential from values around +25 mV to values around 0 mV. Bile salts activated both anionic and cationic channels and did not require the presence of intracellular factors for these effects. We suggest that they act at the membrane level.     

Regional cerebral blood flow during acute hypoxia in individuals susceptible to acute mountain sickness

Individuals susceptible to high altitude pulmonary edema show altered pulmonary vascular responses within minutes of exposure to hypoxia. We hypothesized that a similar acute-phase vulnerability to hypoxia may exist in the brain of individuals susceptible to acute mountain sickness (AMS). In established AMS and high altitude cerebral edema, there is a propensity for vasogenic white matter edema. We therefore hypothesized that increased cerebral blood flow (CBF) during acute hypoxia would also be disproportionately greater in white matter (WM) than grey matter (GM) in AMS-susceptible subjects. We quantified regional CBF using arterial spin labeling MRI during 30 min hypoxia (F(I)O(2) = 0.125) in two groups: AMS-susceptible (AMS-S, n = 6) who invariably experienced AMS at altitude, and AMS-resistant (AMS-R, n = 6) who never experienced AMS despite multiple rapid ascents to high altitude. SaO(2) during hypoxia did not differ between groups (AMS-S = 87+/-4%, AMS-R = 89+/-3%, p = 0.3). Steady-state whole-brain CBF increased in hypoxia (p<0.005), but did not differ between groups (normoxia: AMS-S = 42.7+/-14.0 ml/(100 g min), AMS-R = 41.7+/-10.1 ml/(100 g min); hypoxia: AMS-S = 47.8+/-19.5 ml/(100 g min), AMS-R = 48.2+/-10.1 ml/(100 g min), p = 0.65), and cerebral oxygen delivery remained constant. The percent change in CBF did not differ between brain regions or between groups (although absolute CBF change was greater in GM): (GM: AMS-S = 6.1+/-7.7 ml/(100 g min) (10+/-11%), AMS-R = 8.3+/-5.7 ml/(100 g min) (17+/-11%), p = 0.57; WM: AMS-S = 4.3+/-5.1 ml/(100 g min) (12+/-15%), AMS-R = 4.8+/-2.9 ml/(100 g min) (16+/-9%), p = 0.82). CONCLUSION: CBF increases in acute hypoxia, but is not different between WM and GM, irrespective of AMS susceptibility. Acute phase differences in regional CBF during acute hypoxia are not a primary feature of susceptibility to AMS.     

Hypoxia decreases cellular ATP demand and inhibits mitochondrial respiration of a549 cells

Hypoxia inhibits activity and expression of transporters involved in alveolar Na reabsorption and fluid clearance. We studied whether this represents a mechanism for reducing energy consumption or whether it is the consequence of metabolic dysfunction. Oxygen consumption (JO2) of A549 cells and primary rat alveolar type II cells was measured by microrespirometry during normoxia, hypoxia (1.5% O2), and reoxygenation. In both cell types, acute and 24-h hypoxia decreased total JO2 significantly and reoxygenation restored JO2 after 5 min but not after 24 h of hypoxia in A549 cells, whereas recovery was complete in type II cells. In A549 cells under normoxia Na/K-ATPase accounted for approximately 15% of JO2, whereas Na/K-ATPase-related JO2 was decreased by approximately 25% in hypoxia. Inhibition of other ion transporters did not affect JO2. Protein synthesis-related JO2 was not affected by acute hypoxia, but decreased by 30% after 24-h hypoxia. Acute and 24-h hypoxia decreased JO2 of A549 cell mitochondrial complexes I, II, and III by 30-40%. Reoxygenation restored complex I activity after acute hypoxia but not after 24-h hypoxia. ATP was decreased 30% after 24-h hypoxia, but lactate production rate was not affected. Reduced nicotinamine adenine dinucleotide was slightly elevated in acute hypoxia. Our findings indicate that inhibition of the Na/K-ATPase by hypoxia contributes little to energy preservation in hypoxia. It remains unclear to what extent hypoxic inhibition of mitochondrial metabolism affects ATP-consuming processes.     

The ventilatory response to hypoxia in the anesthetized rat

Inhibitory postsynaptic membrane channels which are activated by glycine were investigated by means of the noise analysis technique. Dose-response curves were obtained for gamma-aminobutyric acid (GABA) in the presence and in the absence of glycine, and it was concluded that GABA and glycine are likely to activate the same receptors. However, glycine proved to have a very low affinity for the inhibitory postsynaptic receptors; this affinity was smaller than that of GABA by a factor of 1 . 10(3)-2 . 10(3). The mean open time tau of the postsynaptic Cl- channels activated by glycine at E = -100 mV and E = -60 mV membrane potentials were tau = 6.1 ms +/- 1.5 ms and tau = 17.7 ms +/- 2.2 ms, respectively. These values are in agreement with the tau obtained by activation with GABA (Dudel et al. 1980); however, on activation by glycine the potential dependence of tau was larger by a factor of 1.35. At E = -100 mV the conductance gamma of glycine-operated channels was about 3 pS which is a third of the respective conductance elicited by GABA. In the presence of high concentrations of glycine (0.1-0.5 mol/l) spontaneous inhibitory postsynaptic currents (sIPSCs) and 'giant' spontaneous inhibitory postsynaptic currents (gsIPSCs) were observed. Furthermore at high concentrations of glycine an additional glycine-induced noise component was found in the power spectra of current fluctuations at higher frequencies. It was concluded that this spectral component resulted from the closing of otherwise open K+ channels, as has been observed already on application of GABA (Dudel and Finger 1980). The mean duration of the low conductance state was tau- = 2.2 ms +/- 0.9 ms and the conductance decrease gamma- coupled to this process was estimated to be about 3 pS. In Na+ free- and Ca2+-enriched bathing solutions the glycine-induced conductances gamma and gamma- were reduced by a factor of about 1.7 while tau and tau- remained unchanged. The decrease in gamma and gamma- was most likely effected by the increase in concentration of divalent cations.     

Measurement of the conductance of the sodium channel from current fluctuations at the node of Ranvier.

Single myelinated nerve fibres of Rana esculenta were investigated under voltage clamp conditions at 13 degrees C. Fluctuations of steady-state membrane current were measured during the last 152 msec of 190-225 msec pulses depolarizing the membrane by 8-48 mV. Noise power spectral densities were calculated in the frequency range of 6-6-6757 Hz. 2. External application of 150 nM tetrodotoxin (TTX) and/or 10 mM tetraethylammonium (TEA) ion reduced the current fluctuations. The difference of current noise spectra measured in the presence and absence of TTX (TEA) was not changed by the presence of TEA (TTX) during both measurements, and was taken as the spectrum of the Na (K) current fluctuations. 3. Residual current noise during application of both TTX and TEA was, except for some excess noise at the low and high frequency ends of the spectrum, similar to the noise measured from a passive nerve model and could be understood in terms of Nyquist noise of the known resistances and the amplifier noise. 4. Na current fluctuation spectra were interpreted as the sum N/f+SNa(f) where SNa(F) represents the spectrum expected for a set of equal, independent Na channels with only two conductance states (open or closed) which follow Hodgkin-Huxley kinetics. With values of hinfinity, tauh and minfinity measured from macroscopic Na currents, the measured spectra were fitted well by optimizing N, SNa(0) and taum. Values of taum obtained by this method were in fair agreement with values found from macroscopic currents. 5. The 1/f component of Na current noise was roughly proportional to the square of the steady-state Na current, I2. The mean value of N/I2 was (1-1 +/- 0-3) X 10(-4). 6. The current carried by a single Na channel was calculated from fitted spectra and steady-state Na currents measured simultaneously with the current fluctuations. The single channel conductance gamma normalized to zero absolute membrane potential was calculated. The average gamma from twelve measurements at depolarizations of 8-40 mV was 7-9 +/- 0-9 pS (S.E. of mean). The apparent value of gamma was smallest with small depolarizations. Variations of the assumed kinetic properties of the model did not drastically affect the single channel conductance. 7. External application of 0-1 mM-Ni ion lengthened taum in the macroscopic currents and in the fluctuation spectra and enhanced both the steady-state Na current and the current fluctuations. In Ni-treated nodes gamma was smaller than in normal nodes.     

Cerebral and muscle tissue oxygenation in acute hypoxic ventilatory response test

Eight men were exposed to progressive isocapnic hypoxia for 10 min to test the hypothesis that (i) cerebral and muscle tissue would follow similar deoxygenation profiles during an acute hypoxic ventilatory response (AHVR) test; and (ii) strong cerebrovascular responsiveness to hypoxia would be related to attenuated cerebral deoxygenation. End-tidal O(2) concentration was reduced from normoxia (approximately 102 mmHg) to approximately 45 mmHg while arterial oxygen saturation (SpO2 %) declined from 98+/-1% to 77+/-7% (P<0.001). Near-infrared spectroscopy (NIRS)-derived local cerebral tissue (frontal lobe) deoxyhemoglobin increased 5.55+/-2.22 microM, while oxyhemoglobin and tissue oxygenation index decreased 2.57+/-1.99 microM and 6.2+/-3.4%, respectively (all P<0.001). In muscle (m. vastus lateralis) the NIRS changes from the initial normoxic level were non-significant. Cerebral blood velocity (V(mean), transcranial Doppler) in the middle cerebral artery increased from 53.4+/-10.4 to 60.6+/-11.6 cms(-1) (P<0.001). In relation to the decline in SpO2 % the mean rate of increase of V(mean) and AHVR were 0.33+/-0.19 cms(-1)%(-1) and 0.52+/-0.20l min(-1)%(-1), respectively. We conclude that cerebral, but not muscle, tissue shows changes reflecting a greater deoxygenation during acute hypoxia. However, the changes in NIRS parameters were not related to cerebrovascular responsiveness or ventilatory chemosensitivity during graded hypoxia.     

Hypoxia impairs the arousal response to external resistive loading and airway occlusion during sleep

STUDY OBJECTIVES: Sustained hypoxia is a neurocognitive depressant, which has been shown to impair respiratory load sensation. Hypoxia has also been shown to impair arousal in animal models, but the effects of sustained hypoxia on arousal in humans have not been studied. The aim of this study was to assess the effects of sustained hypoxia on arousal from sleep in normal subjects. DESIGN: Twelve normal male subjects (age, 24.3 +/- 1.2 years; body mass index, 24.8 +/- 1.4 kg/m2) were studied during stable stage 2 non-rapid eye movement sleep on 2 separate nights 1 week apart. SETTING: Sleep physiology laboratory. PARTICIPANTS: Normal healthy volunteers. Interventions: Arousal responses to external resistive loads (18 cm H2O x L(-1) x sec(-1)) and occlusions were compared during room-air breathing following sustained normoxia and isocapnic hypoxia (SaO2 approximately 85%). Measurements and Results: Time to arousal and minimum esophageal pressure preceding arousal were measured. Time to arousal was significantly increased following hypoxia compared with normoxia for resistive loads (24.6 + 4.4 seconds vs. 12.6 +/- 1.9 seconds, p = .007) but not occlusions. Minimum esophageal pressure prior to arousal was more negative following hypoxia for both external loads (-16.8 +/- 1.2 vs. -13.5 +/- 1.3 cm H2O, p = .035) and occlusions (-19.6 +/- 2.2 vs. -15.1 +/- 1.5 cm H2O, p = .029). CONCLUSIONS: We conclude that sustained isocapnic hypoxia delays arousal to inspiratory loading during sleep and increases the respiratory arousal threshold. This has implications for disorders characterized by sustained nocturnal hypoxia, such as neuromuscular weakness, chronic obstructive pulmonary disease, obesity-hypoventilation syndrome, and severe obstructive sleep apnea.     

Subconductance states of the cardiac KATP channel revealed by partial block with glybenclamide

Currents carried by single ATP-sensitive potassium channels (K(ATP) channels) were recorded in membrane patches isolated from adult rat ventricular myocytes. Channel currents were blocked completely by ATP at millimolar concentrations and by glybenclamide at micromolar concentrations. However, at lower glybenclamide concentrations (1-1000 nM), a partial block, manifest as a subconductance state, was often seen. At concentrations of 100-300 nM the mean size of the subconductance state was 33+/-2.7 pS (175 mM potassium in the pipette; n=13). The size of the conductance substate varied slightly with the concentration of glybenclamide, (42 pS at 1 nM, 34 pS at 100 nM and 31 pS at 1 microM), while the open time of the subconductance state decreased with increasing glybenclamide concentration (n=4). ATP (4 mM) completely blocked both the main conductance state of the channel and the subconductance state induced by glybenclamide. Submaximal concentrations of ATP also appeared to induce subconductance states, but these could not be resolved into discrete conductance levels. The observation that subconductance states can be induced by low concentrations of glybenclamide may have implications for models of how the binding of glybenclamide is translated into closure of the Kir6.2 pore.     

Large-conductance calcium-activated potassium channels in neonatal rat intracardiac ganglion neurons

The properties of single Ca2+-activated K+ (BK) channels in neonatal rat intracardiac neurons were investigated using the patch-clamp recording technique. In symmetrical 140 mM K+, the single-channel slope conductance was linear in the voltage range -60/+60 mV, and was 207+/-19 pS. Na+ ions were not measurably permeant through the open channel. Channel activity increased with the cytoplasmic free Ca2+ concentration ([Ca2+]i) with a Hill plot giving a half-saturating [Ca2+] (K0.5) of 1.35 microM and slope of approximately equals 3. The BK channel was inhibited reversibly by external tetraethylammonium (TEA) ions, charybdotoxin, and quinine and was resistant to block by 4-aminopyridine and apamin. Ionomycin (1-10 microM) increased BK channel activity in the cell-attached recording configuration. The resting activity was consistent with a [Ca2+]i <100 nM and the increased channel activity evoked by ionomycin was consistent with a rise in [Ca2+]i to > or =0.3 microM. TEA (0.2-1 mM) increased the action potential duration approximately equals 1.5-fold and reduced the amplitude and duration of the afterhyperpolarization (AHP) by 26%. Charybdotoxin (100 nM) did not significantly alter the action potential duration or AHP amplitude but reduced the AHP duration by approximately equals 40%. Taken together, these data indicate that BK channel activation contributes to the action potential and AHP duration in rat intracardiac neurons.     

β2-Adrenergics in hypoxia desensitize receptors but blunt inhibition of reabsorption in rat lungs

Alveolar edema and decreased inspired Po(2) decrease the oxygen supply to alveolar epithelia, impairing β(2)-adrenergic receptor (β2AR) signaling and alveolar reabsorption. β2AR agonists potently stimulate alveolar reabsorption. Thus, hypoxia impairs a major defense mechanism that provides protection from alveolar edema. Because in vivo data on the combined effects of prolonged hypoxia and β2AR agonist treatment on β2AR signaling are sparse, we tested whether in vivo hypoxia augments the inactivation of β2AR during prolonged stimulation. Rats were exposed to normoxia (N) and hypoxia (8% O(2); H), and were also treated with terbutaline (T; 2.5 mg/kg, intraperitoneal, twice daily) or saline (S) for 4 days. β2AR signaling was studied in alveolar epithelial (ATII) cells and in whole-lung tissue from treated rats. The terbutaline-stimulated formation of cyclic adenosine monophosphate was decreased by approximately 40% in whole lung and in ATII cells of NT, HS, and HT. The effects were not additive. The β2AR number was increased in HS, but decreased in NT and HT. Treatment increased the G-protein-coupled receptor kinase 2 protein in the plasma membranes of ATII cells, but did not affect G proteins. In vivo hypoxia significantly decreased total and amiloride-sensitive alveolar fluid reabsorption, which was prevented by acute alveolar treatment and 4 days of systemic terbutaline treatment. The αENaC (subunit of epithelial Na channels) protein in plasma membranes was increased in HT, without effects on mRNA. These results indicate that prolonged alveolar hypoxia and treatment with terbutaline impaired β2AR signaling in alveolar epithelia and in whole lungs, and this signaling was not further impaired by hypoxia. Despite impaired β2AR signaling, treatment with terbutaline for 4 days prevented the inhibition of alveolar reabsorption caused by in vivo hypoxia.