Andrzej Trzebski

Carotid chemoreceptor discharge responses to hypoxia and hypercapnia in normotensive and spontaneously hypertensive rats

The carotid chemoreceptor discharge responses to hypoxia and hypercapnia were quantitatively compared between normotensive (NTR) and spontaneously hypertensive rats (SHR). For this purpose we recorded afferent mass discharges from the carotid sinus nerve (CSN) at various levels of end-tidal O2 and CO2 concentrations (FetO2, FetCO2 (%)) in the urethane-anesthetized, vagotomized and artificially ventilated rats. The CSN chemoreceptor discharge was evaluated by subtracting the small activity remaining in acute hyperoxia (chemoreceptor inactivation), which was estimated as baroreceptor in origin, from the large total CSN activity. The CSN chemoreceptor discharges at various levels of FetO2 or FetCO2 were expressed as the percent of control activity measured in normoxic and normocapnic conditions (FetO2, 15-16%; FetCO2, 4.5-5.1%). There was an exponential increase in the CSN chemoreceptor discharge as FetO2 was decreased from hyperoxic to various hypoxic levels (maximally 6%) at a maintained FetCO2 (normocapnia). The relationship between the CSN chemoreceptor discharge and the hypoxic stimulus was quantitatively assessed by the regression analysis using an exponential function. Exponential increases in the CSN chemoreceptor discharge by hypoxia and the parameters in the exponential function reflecting the sensitivity to hypoxia were significantly higher in the SHR than in the NTR, which indicated a high carotid chemoreceptor discharge response to hypoxia in the SHR. The CSN chemoreceptor discharge was increased linearly by increasing the FetCO2 from the normocapnic level up to about 10% at a maintained FetO2 (normoxia). Increases in discharge produced by severe hypercapnia were, however, much smaller than that caused by hypoxia. The slope of the CO2 stimulus-CSN chemoreceptor discharge response line was almost the same in NTR and SHR. The results demonstrated that the responsiveness of rat carotid chemoreceptor to hypoxia is augmented in the SHR. The role of carotid chemoreceptor afferents in ventilatory reflex responses to hypoxia and their alterations in the SHR are discussed.

Autonomic nerve and cardiovascular responses to changing blood oxygen and carbon dioxide levels in the rat

Contribution of autonomic nervous system activity to the heart rate and blood pressure responses during chemoreceptor excitations by systemic hypoxia and hypercapnia and to hyperoxia and hypocapnia was analyzed in the urethane-anesthetized, artificially ventilated rats. Systemic hypoxia induced a co-activation of two antagonistic nerves: an increase in cardiac sympathetic and in cardiac vagal efferent nerve discharges. Increased heart rate was due to predominance of the cardiac sympathetic over the cardiac vagal activation. In spite of a marked reflex increase in the renal and cardiac sympathetic nerve activities, the local vasodilator effect of hypoxia prevented consistent changes in arterial blood pressure. Bilateral section of the carotid sinus nerves (CSN) mostly abolished autonomic nerve responses and produced a profound decreases in the blood pressure during hypoxia. Hyperoxia elicited a pressor response due to peripheral vasoconstriction with no significant change in the autonomic nerve activities except for a decrease in the cardiac sympathetic nerve discharges. Hypercapnia significantly increased blood pressure and renal nerve sympathetic activity. In contrast to hypoxia, hypercapnia excited cardiac sympathetic and inhibited cardiac vagal activity. This reciprocal effect did not elicit neurogenic cardioacceleration, because it was masked by the local inhibitory action of CO2 on the heart rate. The increase in sympathetic activities and in blood pressure during hypercapnia persisted after bilateral CSN section indicating that the responses were mediated by central rather than by peripheral chemoreceptors. Hypocapnia produced a significant increase in cardiac vagal discharges yet a cardioacceleratory response occurred due to the local effect upon heart rate. The present results indicate that in the rat, autonomic nervous responses differ depending on the type, i.e. hypoxic or hypercapnic, chemoreceptor stimuli. Reflex heart rate and blood pressure responses do not follow the autonomic nerve activities exactly. Circulatory responses are greatly modified by local peripheral effects of hypoxic, hyperoxic, hypocapnic or CO2 stimuli on the cardiovascular system. Species differences characterizing the autonomic nerve responsiveness to chemical stimuli in the rat are described.

Fast inhibitory postsynaptic potentials and responses to inhibitory amino acids of sympathetic preganglionic neurons in the adult cat

Intracellular recordings were obtained from sympathetic preganglionic neurons (SPNs) of the intermediolateral nucleus (IML) in slices of upper thoracic spinal cord of the anesthetized cat. A total of 44 neurons was studied. Single shock stimulation of an area of white matter dorsolateral to the IML, close to the recording electrode (< 0.5 mm), evoked fast IPSPs with rise time of 3.8 ms and 1/2 decay time of 14.7 ms (n = 12). In 17 other cells only fast EPSPs were recorded but, after suppression of the EPSPs by the excitatory amino acid receptor antagonists CNQX (20 microM) and APV (100-250 microM), fast IPSPs were unmasked. The IPSP reversed polarity at -63 mV (-67 mV in the presence of CNQX and APV). The reversal potential shifted to a less negative value when the extracellular chloride concentration was reduced. The IPSP was reversibly abolished by the GABAA receptor antagonist bicuculline in 32% of the cells, by the glycine receptor antagonist strychnine in 47% of the cells and by the combination of the two in 21% of the cells. The IPSP was abolished by TTX (0.5 microM), had constant latency and showed no failures during high frequency stimulation. The IPSP presumably resulted from the excitation of inhibitory axons and/or inhibitory neuron somata with monosynaptic connections to the SPN. Glycine and GABA (1-3 mM) produced hyperpolarization associated with decreased membrane resistance. Sixty-nine percent of cells responded to both agonists, 19% to glycine only and 12% to GABA only. The GABAB agonist baclofen (5 microM) had no effect.(ABSTRACT TRUNCATED AT 250 WORDS)

Is the central inspiratory activity responsible for pCO2-dependent drive of the sympathetic discharge?

Out of 27 cats anesthetized with chloralose-urethane mixture, paralyzed, vagotomized and artificially ventilated, phrenic nerve response to systemic hypercapnia (7-8 vol.% CO2/O2 mixture) was accompanied by an increase in blood pressure and sympathetic discharge in 19 cats. Out of these 19 cats, 12 were totally debuffered and in the remaining 7 cats one carotid sinus nerve was left intact. Single unit activity in the sympathetic cervical nerve and spontaneous mass activity in the cervical, splanchnic, renal sympathetic and phrenic nerves were recorded. Evoked response in the phrenic nerve was produced by electrical stimulation of the descending bulbospinal inspiratory pathways in the midplane area of the medulla or in the ventrolateral cervical spinal cord. Starting from the control mean end-tidal CO2 concentration of 4.7 vol.% (+/- 1.0 S.D.) a progressing hypocapnia was induced by hyperventilation up to the end-tidal CO2 concentration of 1.3-3.2 vol.% (mean 2.4 vol.% +/- 0.5 S.D.) significantly below paCO2 apneic threshold. In chemo- and baroreceptor denervated cats with a pressor and excitatory sympathetic response to hypercapnia, a hypocapnia resulted in a fall of the arterial blood pressure (mean 16.9 mm Hg +/- 7.5 S.D., 2.2 kpa +/- S.D.). With the increasing paCO2 over the period of hypocapnic apnea a pressor and excitatory sympathetic response preceded, in all experiments, the onset of the phrenic nerve rhythmic activity. The difference between paCO2 threshold for the pressor and sympathetic response (35.7 mm Hg +/- 3.6 S.D., 4.7 kpa +/- 0.5 S.D.) and paCO2 threshold for the reappearance of the phrenic nerve rhythmic activity (43.6 mm Hg +/- 2.6 S.D., 5.8 kpa +/- 0.3 S.D.) was highly significant. If apneic hypocapnia was combined with the continuous stimulation of the afferent fibers of the superior laryngeal nerve the CO2 threshold for phrenic rhythmic activity was significantly increased whereas CO2 threshold for the pressor and sympathetic excitatory response remained unchanged. CO2 administration during hypocapnia apnea caused a progressing reduction of the magnitude of the evoked phrenic nerve response. From these findings it is concluded that the central excitatory effect of CO2 on the sympathetic activity may be accomplished in the absence of the rhythmic respiratory activity and independently of the subthreshold tonic inspiratory activity. Pressor and sympathetic excitatory response to CO2 observed during hypocapnic apnea is presumably caused by a neuronal pool different from that responsible for the central inspiratory activity. It is suggested that this CO2 sensitive neuronal mechanism might be involved in the central generation of sympathetic tone.

Inhibition of nitric oxide synthesis potentiates the responsiveness of carotid chemoreceptors to systemic hypoxia in the rat

Carotid sinus nerve afferent activity was recorded in the peripheral end of the cut carotid sinus nerves in rats anesthetized with urethane, paralyzed and artificially ventilated with pure oxygen in order to abolish any resting chemoreceptor activity. Hypoxic stimuli were applied by switching pure oxygen to a nitrogen/oxygen gas mixture in the inspiratory line, reducing end-tidal oxygen concentrations to 10% FETO2, 8% FETO2 and 6% FETO2 respectively. Each stimulus was applied for 60 s and ventilation was switched again to pure oxygen. Increases in the carotid sinus nerve activities were due to chemo- and not to baroreceptor stimulation as arterial blood pressure decreased during hypoxia. After administration of nitric oxide synthase blocker L-NG-nitroarginine methyl ester, 30 mg/kg weight i.v., chemoreceptor excitatory response to all hypoxic stimuli increased significantly. Subsequent administration of L-arginine, 300 mg/kg weight i.v., restored chemoreceptor response to hypoxia to initial magnitude. It is concluded that NO is generated in the carotid body and attenuates chemoreceptor responsiveness in rats in vivo, as reported on isolated carotid bodies in cats in vitro.

Role of the rostral ventrolateral medulla in the generation of synchronized sympathetic rhythmicities in the rat.

In artificially ventilated, paralyzed rats anesthetized with Nembutal or urethane, power density spectral analysis (PDS), using direct FFT algorithm, was used to quantify rhythmicities in the sympathetic cervical and lumbar nerves after bilateral microinjections into rostral ventrolateral medulla (RVLM) of CoCl2 and MgCl2--unspecific synaptic transmission blockers. Later overall sympathetic activity, phrenic nerve discharge, heart rate and arterial blood pressure were recorded. Block of synaptic transmission in RVLM was tested by elimination of sympathoinhibitory or sympathoexcitatory reflex responses to aortic nerve and vagal afferents stimulation respectively. In animals vagotomized with bilateral section of carotid sinus nerve the power in all frequency bands was very significantly reduced to a level not different from that which remained after spinal cord transsection. If carotid baroreceptors were intact, a small peak corresponding to cardiac frequency band persisted. Overall, non-synchronized sympathetic activity and arterial blood pressure increased. All effects were transient, lasted up to 15 min, and could be reproduced several times in one experiment. Respiratory rhythmic activity was unchanged yet respiratory-sympathetic synchronization was abolished. It is concluded that RVLM reticulospinal sympathoexcitatory neurons are responsible for non-synchronized tonic sympathetic activity but are not able to generate synchronized sympathetic rhythms. Synaptic input, presumably inhibitory, either from local neuronal circuits within ventral medulla and/or from other brain stem neuronal circuitries is needed to shape out the flexible pattern of sympathetic oscillations.

Cardiovascular periodicities in healthy humans in the absence of breathing and under reduced chemical drive of respiration

In 20 healthy human subjects (aged 20-24 years) power density spectra in 1-min time series were computed by modified MATLAB procedures from inter-beat heart intervals and systolic and diastolic finger blood pressure. High- (HF), mid- (MF) and low-frequency (LF) peaks (0.16-0.37 Hz, 0.08-0.12 Hz and 0.02-0.05 Hz, respectively) were distinguished in power density spectra (PDS). During 1-min voluntary apneas MF and LF power increased and in 18 out of 20 subjects HF peaks disappeared in PDS. Breath holding performed in hyperoxia or in hyperoxia accompanied by 1-min hyperventilation preceding apnea augmented power at MF and LF range and abolished marginal HF peaks present in the spectrum in two subjects. It is concluded that apneic PDS is characterized by augmented power of MF and LF rhythms, and represents endogenous free running cardiovascular periodicities released from feed-back mechanical, reflex and feed-forward central entrainment to respiratory rhythm.

Non-linear dynamics of cardiovascular system in humans exposed to repetitive apneas modeling obstructive sleep apnea: aggregated time series data analysis

OBJECTIVE Testing the hypothesis that alterations in non-linear dynamics of the cardiovascular variability appear in healthy, awake subjects during voluntary apnea. SUBJECTS AND METHODS Ten young subjects performed 20 apneas 60 s each separated by 1 min free breathing. Inter-beat interval (IBI) measured as RR-interval in ECG, systolic (SYS) and diastolic (DIAS) arterial blood pressure. stroke volume (SV), cardiac output (CO) and total peripheral resistance (TPR) were non-invasively recorded and computed by Portapress 2 system. Correlation dimension according to Grassberger-Proccacia algorithm (CD) and ratio of % determinism versus % recurrence (RDR) computed from recurrence plot according to Webber and Zbiluth [J. Appl. Physiol. 76 (1994) 965] were used as the indices of chaos and complexity. Sequential time series of cardiovascular variables in consecutive 60-s apneas and inter-apnea free breathings were separately windowed and aggregated. CD and RDR of aggregated apneic time series were compared with CD and RDR values of aggregated time series of inter-apnea free breathing, 10 min at rest and 10 min recovery. Reliability of the aggregation method of sequential time series was tested on transformed simulated data generated by Lorenz model. Error in CD and RDR estimation did not exceed 5% and 1%. respectively. RESULTS CD of cardiovascular variables computed from aggregated apneas was significantly reduced and RDR augmented versus control and recovery periods. CD and RDR values of inter-apnea phases were in between those in control and during apneas. Time domain linear variance (SD) was increased during aggregated apneas. CONCLUSIONS Signal dynamics dependent windowing and data aggregation could be a useful tool for non-linear analysis of short repeatable sequential time series; time domain linear variability of the cardiovascular dynamics is augmented while complexity reduced during apneic chemoreceptor stimulation; alterations in non-linear dynamics of cardiovascular variables during apneas in healthy normotensive subjects may suggest an early sign of a mechanism implicated in arterial hypertension in OSAS.

Local cerebral blood flow responses in rats to hypercapnia and hypoxia in the rostral ventrolateral medulla and in the cortex

The effects of hypercapnia and hypoxia on two local cerebral blood flows in the parietal cortex (PC-BF) and rostral ventrolateral medulla (RVLM-BF) were examined using laser Doppler flowmetry in anesthetized rats. Hypercapnia for 45 s duration at the end-tidal CO2 between 5% and 10%, induced by increasing inspiratory CO2, increased both cerebral blood flows and systemic blood pressure in a degree-dependent manner. The response of RVLM-BF was significantly stronger than that of PC-BF. Both cerebral blood flow responses to hypercapnia were not influenced by cutting peripheral chemoreceptor afferent nerves. Hypoxia for 45 s duration at the end-tidal O2 between 12% and 6%, induced by decreasing inspiratory O2, produced an increase of similar magnitude in both RVLM and PC local blood flows in a degree-dependent manner and a decrease in systemic blood pressure. The responses of both PC-BF and RVLM-BF to hypoxia were significantly diminished after cutting peripheral chemoreceptor afferent nerves. It is concluded that: (1) the RVLM-BF is much more sensitive to hypercapnia than the PC-BF; and (2) activation of peripheral arterial chemoreceptors possibly contributes to hypoxia-induced increase in the RVLM-BF and PC-BF.

Respiratory rhythmicity in a split medulla preparation of the cat

Splitting the medulla in the cat resulted in the disappearance of inspiratory activity in the phrenic but not in the recurrent laryngeal nerves with minor changes in respiratory cycle duration. In one animal a complete desynchronization of respiratory rhythms was observed in the opposite recurrent laryngeal nerves. We conclude that each half of the cats brain stem has its own respiratory phase-switching mechanism. Commissural connections are involved in the bilateral synchronization of activity and the mutual excitatory interactions between medullary respiratory neurons.