Responses of neurons in the rostral ventrolateral medulla to whole body rotations: comparisons in decerebrate and conscious cats.

The responses to vestibular stimulation of brain stem neurons that regulate sympathetic outflow and blood flow have been studied extensively in decerebrate preparations, but not in conscious animals. In the present study, we compared the responses of neurons in the rostral ventrolateral medulla (RVLM), a principal region of the brain stem involved in the regulation of blood pressure, to whole body rotations of conscious and decerebrate cats. In both preparations, RVLM neurons exhibited similar levels of spontaneous activity (median of ∼17 spikes/s). The firing of about half of the RVLM neurons recorded in decerebrate cats was modulated by rotations; these cells were activated by vertical tilts in a variety of directions, with response characteristics suggesting that their labyrinthine inputs originated in otolith organs. The activity of over one-third of RVLM neurons in decerebrate animals was altered by stimulation of baroreceptors; RVLM units with and without baroreceptor signals had similar responses to rotations. In contrast, only 6% of RVLM neurons studied in conscious cats exhibited cardiac-related activity, and the firing of just 1% of the cells was modulated by rotations. These data suggest that the brain stem circuitry mediating vestibulosympathetic reflexes is highly sensitive to changes in body position in space but that the responses to vestibular stimuli of neurons in the pathway are suppressed by higher brain centers in conscious animals. The findings also raise the possibility that autonomic responses to a variety of inputs, including those from the inner ear, could be gated according to behavioral context and attenuated when they are not necessary.

Effects on sympathetic activity of 8-OHDPAT and clonidine in cat medullary lateral tegmental field.

This study was designed to test the hypothesis that 8-hydroxy-2-(di-n-propylamino)tetralin (8-OHDPAT) and clonidine reduce sympathetic nerve discharge (SND) and mean arterial pressure (MAP), in part by actions in the medullary lateral tegmental field (LTF). We microinjected these drugs bilaterally into the LTF of baroreceptor-innervated and -denervated cats anesthetized with Dial-urethane. Neither drug altered SND (as quantified by using power spectral analysis) or MAP when injected into the LTF of baroreceptor-denervated cats. However, cardiac-related power in SND was significantly increased to 148 +/- 12 (mean +/- SE) and 149 +/- 5% of control by microinjections of 8-OHDPAT (n = 5) and clonidine (n = 5), respectively, in baroreceptor-innervated cats whose MAP was kept constant; there was no change in 0- to 6-Hz power or total power. SND was significantly reduced by microinjection of these drugs into the rostral ventrolateral medulla of baroreceptor-innervated and -denervated cats. In conclusion, although 8-OHDPAT and clonidine did not reduce SND when injected into the LTF, they acted in this region to facilitate baroreceptor reflex control of SND, as evidenced by a selective increase in cardiac-related power.

The role of the medullary lateral tegmental field in the generation and baroreceptor reflex control of sympathetic nerve discharge in the cat.

Data from experiments with single neuron recordings as well as central microinjections of N-methyl-D-aspartate (NMDA) and non-NMDA excitatory amino receptor antagonists that have led to a model of central sympathetic pathways that includes synaptic relays in the medullary lateral tegmental field (LTF) of the cat are summarized. Evidence is presented that (1) the LTF contains a population of tonically active sympathoexcitatory neurons that drive rostral ventrolateral medullary neurons, (2) blockade of non-NMDA receptors in the LTF significantly reduces basal levels of sympathetic nerve discharge (SND) and mean arterial pressure in baroreceptor-denervated cats, and (3) blockade of NMDA-mediated neurotransmission in the LTF prevents baroreceptor reflex control of SND. Thus, LTF neurons play an important role in the generation and baroreceptor reflex control of SND in the cat.

Sympathetic nerve and cardiovascular responses to chemical activation of the midbrain defense region.

The changes in mean arterial pressure (MAP), renal (RBF) and femoral (FBF) blood flows, and inferior cardiac (CN) and vertebral nerve (VN) sympathetic nerve discharges (SND) produced by chemical activation (D,L-homocysteic acid) of the midbrain periaqueductal gray (PAG) were compared in baroreceptor-denervated and -innervated cats anesthetized with urethan. Defenselike cardiovascular responses in both states were similar in magnitude and consisted of increased MAP and FBF and decreased RBF; however, the nerve responses differed. In baroreceptor-denervated cats, PAG activation increased CN 10-Hz activity, decreased VN 10-Hz activity, and lengthened the CN-VN phase angle. In baroreceptor-innervated cats in which the rhythm in SND was cardiac related, PAG activation increased CN activity, but VN activity and the CN-VN phase angle were unchanged. These results demonstrate that chemical activation of PAG neurons induces differential patterns of sympathetic outflow generally consistent with accompanying defenselike cardiovascular responses. However, the mechanisms responsible for the changes in 10-Hz and cardiac-related SND appear to be different.

Long-term correlations in the spike trains of medullary sympathetic neurons.

Fano factor analysis was used to characterize the spike trains of single medullary neurons with sympathetic nerve-related activity in cats that were decerebrate or anesthetized with Dial-urethan or urethan. For this purpose, values (Fano factor) of the variance of the number of extracellularly recorded spikes divided by the mean number of spikes were calculated for window sizes of systematically varied length. For window sizes < or =10 ms, the Fano factor was close to one, as expected for a Bernoulli process with a low probability of success. The Fano factor dipped below one as the window size approached the shortest interspike interval (ISI) and reached its nadir at window sizes near the modal ISI. The extent of the dip reflected the shape (skewness) of the ISI histogram with the dip being smallest for the most asymmetric distributions. Most importantly, for a wide range of window sizes exceeding the modal ISI, the Fano factor curve took the form of a power law function. This was the case independent of the component (cardiac related, 10 Hz, or 2--6 Hz) of inferior cardiac sympathetic nerve discharge to which unit activity was correlated or the medullary region (lateral tegmental field, raphe, caudal and rostral ventrolateral medulla) in which the neuron was located. The power law relationship in the Fano factor curves was eliminated by randomly shuffling the ISIs even though the distribution of the intervals was unchanged. Thus the power law relationship arose from long-term correlations among ISIs that were disrupted by shuffling the data. The presence of long-term correlations across different time scales reflects the property of statistical self-similarity that is characteristic of fractal processes. In most cases, we found that mean ISI and variance for individual spike trains increased as a function of the number of intervals counted. This can be attributed to the clustering of long and short ISIs, which also is an inherent property of fractal time series. We conclude that the spike trains of brain stem sympathetic neurons have fractal properties.

Coupling of arterial pulse and 10-Hz rhythm in sympathetic nerve discharge.

We used time series analysis to characterize the relationships among the arterial pulse (AP) and the cardiac-related and 10-Hz rhythms in sympathetic nerve discharge (SND) of urethane-anesthetized cats. We found that 10-Hz activity was more tightly coupled to the AP than to the cardiac-related rhythm. These data support the view that the dynamic coupling of AP and the 10-Hz rhythm in SND involves a direct influence of baroreceptor activity on the 10-Hz oscillator.

"Rapid" rhythmic discharges of sympathetic nerves: sources, mechanisms of generation, and physiological relevance.

Like virtually all other physiological control systems, the sympathetic nervous system controlling cardiovascular function is characterized by the presence of rhythmic activity. These include slow rhythms with frequencies at or below that of the respiration and rapid rhythms with frequencies at or above that of the heart beat. The rapid rhythms are the subject of this review. The specific questions entertained are as follows: (1) Are the rapid cardiac-related and 10-Hz rhythms inherent to central sympathetic networks, or are they imposed on sympathetic nerve discharge (SND) by extrinsic periodic inputs? (2) Does basal SND arise from an anatomically circumscribed "vasomotor center" composed of pacemaker neurons in the rostral ventrolateral medulla or from an anatomically distributed network oscillator composed of different types of brainstem neurons, none of which necessarily have intrinsic pacemaker properties? (3) Are the rapid rhythms generated by single circuits or by systems of coupled oscillators, each with a separate target? (4) Are the rapid rhythms in SND simply by-products of the sympathetic generating mechanisms, or do they subserve selective and special functions, such as the formulation of differential patterns of spinal sympathetic outflow that support particular behaviors? The controversial aspects of these issues and the state-of-the-art analytical methods used to study them are stressed in this review.

Modes of baroreceptor-sympathetic coordination.

We tested the hypothesis that the cardiac-related rhythm in sympathetic nerve discharge (SND) results from the forcing of a central oscillator to the frequency of the heart beat by pulse-synchronous baroreceptor afferent nerve activity. For this purpose, time series analysis was used to examine the phase relations between the brachial arterial pulse (AP) and cardiac-related activity recorded from the postganglionic inferior cardiac sympathetic nerve (CN) in urethan-anesthetized cats. Specifically, we made cycle-by-cycle measurements of peak systolic blood pressure, heart period, CN burst amplitude, and the phase angle (and corresponding interval) between peak systole and the next peak of CN activity. As the steady-state level of systolic blood pressure was raised by increasing the rate of a constant intravenous infusion of phenylephrine, we observed transitions from no phase-locking of CN activity to the AP to either phase-locking of variable strength or phase walk through part of the cardiac-cycle on the time scale of respiration. Phase walk is defined as a progressive and systematic change in the phase lag of cardiac-related CN activity relative to peak systole. Raising blood pressure strengthened phase-locking and either increased or decreased the mean interval between peak systole and the next peak of CN activity even when the change in heart period was small. CN burst amplitude and the interval between peak systole and the next peak of CN activity were inversely related, but the strength of the relationship varied considerably with experimental conditions. The relationship was strongest during phase walk. Step-wise increases in blood pressure induced by abdominal aortic obstruction led to an abrupt increase in the phase lag of CN activity relative to peak systole even when heart rate was not changed. We refer to such changes as sharp phase transitions that are a general property of dynamical nonlinear systems. The results support the view that the cardiac-related rhythm in SND is a forced nonlinear oscillation rather than the consequence of periodic inhibition of randomly generated activity.

Differential pattern of spinal sympathetic outflow in response to stimulation of the caudal medullary raphe.

In urethan-anesthetized cats, frequency domain analysis was used to explore the mechanisms of differential responses of inferior cardiac (CN), vertebral (VN), and renal (RN) sympathetic nerves to electrical stimulation of a discrete region of the medullary raphe (0-2 mm caudal to the obex). Raphe stimulation in baroreceptor-denervated cats at frequencies (7-12 Hz) that entrained the 10-Hz rhythm in nerve activity decreased CN and RN activities but increased VN activity. The reductions in CN and RN discharges were associated with decreased low-frequency (

Defenselike patterns of spinal sympathetic outflow involving the 10-Hz and cardiac-related rhythms.

Frequency- and time-domain analyses were used to compare the effects of stimulation of the defense region of the midbrain periaqueductal gray (PAG) on the 10-Hz and cardiac-related discharges of sympathetic nerves with different cardiovascular targets. In baroreceptor-denervated cats anesthetized with urethan, PAG stimulation at frequencies equal to or higher (up to 25 Hz) than that of the free-running 10-Hz rhythm produced an immediate and sustained decrease in vertebral sympathetic nerve (VN) 10-Hz activity but increased the 10-Hz discharges of the inferior cardiac (CN) and renal (RN) nerves. In baroreceptor-innervated cats, VN cardiac-related activity was initially unchanged by high-frequency (25-Hz) PAG stimulation, or it increased along with that in the CN and RN. Later, during high-frequency PAG stimulation, when the rise in blood pressure approached its peak, VN cardiac-related activity usually was reduced below control level. At this time, the increases in CN and RN cardiac-related discharges were largely sustained. The cardiac-related discharges of the three nerves were unaffected by PAG stimulation at frequencies just below or just above that of the heartbeat. We conclude that the defenselike pattern of spinal sympathetic outflow involving the 10-Hz rhythm is different in mechanism and character from that involving the cardiac-related rhythm.

Medullary lateral tegmental field: an important source of basal sympathetic nerve discharge in the cat.

We used blockade of excitatory amino acid (EAA) neurotransmission in the medullary lateral tegmental field (LTF) and rostral ventrolateral medulla (RVLM) to assess the roles of these regions in the control of inferior cardiac sympathetic nerve discharge (SND) and mean arterial pressure (MAP) in urethan-anesthetized, baroreceptor-denervated cats. Bilateral microinjection of a non-N-methyl-D-aspartate (NMDA)-receptor antagonist [1,2,3, 4-tetrahydro-6-nitro-2,3-dioxobenzo-[f]quinoxaline-7-sulfonamide (NBQX)] into the LTF significantly decreased SND to 46 +/- 4% of control (as demonstrated with power-density spectral analysis) and MAP by 16 +/- 6 mmHg. In contrast, bilateral microinjection of an NMDA-receptor antagonist [D(-)-2-amino-5-phosphonopentanoic acid (D-AP5)] into the LTF did not decrease SND or MAP. These results demonstrate that the LTF is an important synaptic relay in the pathway responsible for basal SND in the cat. Bilateral microinjection of NBQX or D-AP5 into the RVLM significantly decreased power in SND to 48 +/- 5 or 61 +/- 5% of control, respectively, and reduced MAP by 15 +/- 2 or 8 +/- 4 mmHg, respectively. These data indicate that EAA-mediated synaptic drive to RVLM-spinal sympathoexcitatory neurons accounts for a significant component of their basal activity.

Medullary lateral tegmental field: an important synaptic relay in the baroreceptor reflex pathway of the cat.

This study was designed to test the hypothesis that the medullary lateral tegmental field (LTF) is an important synaptic relay in the baroreceptor reflex pathway controlling sympathetic nerve discharge (SND) of urethan-anesthetized cats. We determined the effects of blockade of excitatory amino acid-mediated neurotransmission in the LTF on three indexes of baroreceptor reflex function: cardiac-related power in SND, strength of linear correlation (coherence value) of SND to the arterial pulse (AP), and inhibition of SND during increased arterial pressure produced by abrupt obstruction of the abdominal aorta. Bilateral microinjection of D-(-)-2-amino-5-phosphonopentanoic acid, an N-methyl-D-aspartate (NMDA) receptor antagonist, abolished cardiac-related power and coherence of SND to the AP, and it prevented inhibition of SND during aortic obstruction. These data support the view that NMDA receptor-mediated neurotransmission in the LTF is critical for baroreceptor reflex control of SND. Bilateral microinjection of 1,2, 3,4-tetrahydro-6-nitro-2,3-dioxobenzo-[f]-quinoxaline-7-sulfonamid e, a non-NMDA receptor antagonist, decreased cardiac-related power and total power in the 0- to 6-Hz band of SND; however, the AP-SND coherence value remained high, and inhibition of SND during aortic obstruction was preserved. These data imply that non-NMDA receptor-mediated neurotransmission in the LTF is involved in setting the level of excitatory drive to sympathetic nerves.

Differential patterns of spinal sympathetic outflow involving a 10-Hz rhythm.

Time and frequency domain analyses were used to examine the changes in the relationships between the discharges of the inferior cardiac (CN) and vertebral (VN) postganglionic sympathetic nerves produced by electrical activation of the midbrain periaqueductal gray (PAG) in urethan-anesthetized, baroreceptor-denervated cats. CN-VN coherence and phase angle in the 10-Hz band served as measures of the coupling of the central oscillators controlling these nerves. The 10-Hz rhythm in CN and VN discharges was entrained 1:1 to electrical stimuli applied to the PAG at frequencies between 7 and 12 Hz. CN 10-Hz discharges were increased, and VN 10-Hz discharges were decreased when the frequency of PAG stimulation was equal to or above that of the free-running rhythm. In contrast, stimulation of the same PAG sites at lower frequencies increased, albeit disproportionately, the 10-Hz discharges of both nerves. In either case, PAG stimulation significantly increased the phase angle between the two signals (VN 10-Hz activity lagged CN activity); coherence values relating their discharges were little affected. However, the increase in phase angle was significantly more pronounced when the 10-Hz discharges of the two nerves were reciprocally affected. Importantly, partialization of the phase spectrum using the PAG stimuli did not reverse the change in CN-VN phase angle. This observation suggests that the increase in the CN-VN phase angle reflected changes in the phase relations between coupled oscillators in the brain stem rather than the difference in conduction times to the two nerves from the site of PAG stimulation. In contrast to the effects elicited by PAG stimulation, stimulation of the medullary lateral tegmental field induced uniform increases in the 10-Hz discharges of the two nerves and no change in the CN-VN phase angle. Our results demonstrate that changes in the phase relations among coupled brain stem 10-Hz oscillators are accompanied by differential patterns of spinal sympathetic outflow. The reciprocal changes in CN and VN discharges produced by PAG stimulation are consistent with the pattern of spinal sympathetic outflow expected during the defense reaction.

Human brain alpha rhythm: nonlinear oscillation or filtered noise?

The mechanism for generation of the alpha rhythm is controversial. In the current study, analysis in the time and frequency domains revealed that the alpha rhythm recorded from the scalp overlying the human occipital cortex can be entrained to the second or third harmonic of low frequency light flashes. These results support the view that the alpha rhythm is generated by a nonlinear oscillator rather than a narrow-band transmission system acting as a filter.

Rostral dorsolateral pontine neurons with sympathetic nerve-related activity.

Spike-triggered averaging, arterial pulse-triggered analysis, and coherence analysis were used to classify rostral dorsolateral pontine (RDLP) neurons into groups whose naturally occurring discharges were correlated to only the 10-Hz rhythm (n = 29), to only the cardiac-related rhythm (n = 15), and to both rhythms (n = 15) in inferior cardiac sympathetic nerve discharge (SND) of urethan-anesthetized cats. Most of the neurons with activity correlated to only the cardiac-related rhythm were located medial to the other two groups of neurons. The firing rates of most RDLP neurons with activity correlated to only the 10-Hz rhythm (9 of 12) or both rhythms (7 of 8) were decreased during baroreceptor reflex-induced inhibition of SND produced by aortic obstruction; thus, they are presumed to be sympathoexcitatory. The firing rates of four of seven RDLP neurons with activity correlated to only the cardiac-related rhythm increased during baroreceptor reflex activation; thus, they may be sympathoinhibitory. We conclude that the RDLP contains a functionally heterogeneous population of neurons with sympathetic nerve-related activity. These neurons could not be antidromically activated by stimulation of the thoracic spinal cord.

Classification of caudal ventrolateral pontine neurons with sympathetic nerve-related activity.

This study was designed to answer three questions concerning caudal ventrolateral pontine (CVLP) neurons whose naturally occurring discharges are correlated to sympathetic nerve discharge (SND). 1) What are the proportions of CVLP neurons that have activity correlated to both the cardiac-related and 10-Hz rhythms in SND, to only the 10-Hz rhythm, and to only the cardiac-related rhythm? 2) Do CVLP neurons with activity correlated to the cardiac-related and/or 10-Hz rhythm in SND subserve a sympathoexcitatory or sympathoinhibitory function? 3) Do CVLP neurons with activity correlated to the cardiac-related and/or 10-Hz rhythm in SND project to the thoracic spinal cord? To address these issues we recorded from 476 CVLP neurons in 24 urethan-anesthetized cats. Spike-triggered averaging, arterial pulse-triggered analysis, and coherence analysis revealed that the discharges of 66 of these neurons were correlated to inferior cardiac postganglionic SND. For 39 of these neurons, we were able to determine whether their discharges were correlated to one or both rhythms. The results showed that the CVLP contained a heterogeneous population of neurons with sympathetic nerve-related activity. The discharges of 21 neurons were correlated to both the 10-Hz and cardiac-related rhythms in SND, 9 neurons had activity correlated to only the 10-Hz rhythm, and 9 neurons had activity correlated to only the cardiac-related rhythm. The firing rates of CVLP neurons with activity correlated to both rhythms or to only the 10-Hz rhythm were decreased during the inhibition of SND induced by baroreceptor reflex activation (rapid obstruction of the abdominal aorta). These neurons are presumed to exert sympathoexcitatory actions. The time-controlled collision test verified that 11 of 12 CVLP neurons with activity correlated to both rhythms were antidromically activated by stimulation of the first thoracic segment of the spinal cord. Antidromic mapping at this level showed that the site requiring the least stimulus current to elicit the longest latency response (nearest the terminal) was in the vicinity of the intermediolateral nucleus (IML). In contrast, only 1 of 13 CVLP neurons with activity correlated to only one of the rhythms in SND could be antidromically activated by spinal stimulation. These data demonstrate for the first time that there is a direct pathway from the CVLP to the IML that is comprised almost exclusively of sympathoexcitatory neurons whose discharges are correlated to both the 10-Hz and cardiac-related rhythms in SND.

Nonlinear interactions of slow and rapid rhythms in sympathetic nerve discharge.

We used bispectral analysis to characterize the nonlinear interactions of the respiratory-related (RR), cardiac-related (CR), or 10-Hz rhythms in sympathetic nerve discharge (SND) of urethan-anesthetized cats. Bispectral analysis investigates relationships among frequency triples in the same signal (inferior cardiac postganglionic SND) where the third frequency is the sum of the other two due to quadratic nonlinear coupling. Coupling of the RR and CR rhythms leading to the generation of new components (i.e., modulated frequencies) in SND occurred in 84% of the total cases, whereas the incidence was 71% for the RR and 10-Hz rhythms. The occurrence of such nonlinear interactions implies that the RR, CR, and 10-Hz rhythms are carried to common targets by the same postganglionic sympathetic neurons. Furthermore, we suggest that nonlinear interactions leading to the generation of new frequencies in SND may affect end-organ function beyond the level expected in simple cases of linear superposition of the primary rhythms. This suggestion is supported by our observation that strong coupling of the RR and CR rhythms resulted in appreciable power at the modulated frequencies.

Paradoxical sympathetic nerve response to baroreceptor reflex activation.

Spectral analysis revealed an enhancement of cardiac-related postganglionic sympathetic nerve discharge (SND) in response to elevated blood pressure in cats. Most of the enhancement occurred at blood-pressure levels above which coherence of SND to the arterial pulse at the frequency of the heart beat became maximal. This raises the possibility that the enhancement is due to mechanisms other than improved phase locking of SND to pulse-synchronous baroreceptor afferent nerve activity.

Pontine neurons are elements of the network responsible for the 10-Hz rhythm in sympathetic nerve discharge.

The current study was designed to test the hypothesis that pontine neurons are elements of the network responsible for the 10-Hz rhythm in sympathetic nerve discharge (SND). The first series of experiments tested whether chemical inactivation of neurons in the rostral dorsolateral pons (RDLP) or caudal ventrolateral pons (CVLP) affected inferior cardiac postganglionic SND of urethan-anesthetized cats. Muscimol microinjections into either region eliminated the 10-Hz rhythm in SND, supporting the view that pontine neurons are involved in the expression of this rhythm. Additional experiments were designed to determine if pontine neurons have activity correlated to the 10-Hz rhythm in SND or whether they merely provide a tonic (nonrhythmic) driving input to the rhythm generator. Coherence analysis revealed that local field potentials recorded from the RDLP or CVLP had a 10-Hz component that was significantly correlated to SND. Also, spike-triggered averaging and coherence analysis showed that the naturally occuring discharges of individual RDLP or CVLP neurons were correlated to the 10-Hz rhythm in SND. Taken together, these data support the hypothesis that RDLP and CVLP neurons are essential for the expression of the 10-Hz rhythm in SND and that they are elements of or receive input from the rhythm generator.

Coupled oscillators account for the slow rhythms in sympathetic nerve discharge and phrenic nerve activity.

Phase-locked slow rhythms in sympathetic nerve discharge (SND) and phrenic nerve activity (PNA) are generally thought to arise from a common brain stem "cardiorespiratory" oscillator. The results obtained in vagotomized and baroreceptor-denervated cats anesthetized with pentobarbital sodium do not support this view. First, partial coherence analysis revealed that the discharges of pairs of sympathetic nerves remained correlated at the frequency of the central respiratory cycle after mathematical removal of the portion of these signals common to PNA. The residual coherence suggests that the slow rhythm in SND is dependent on central mechanisms in addition to those responsible for rhythmic PNA. Second, the rhythms in SND and PNA became coupled in a 2:1 relationship during either moderate systemic hypocapnia or hypercapnia. Third, the slow rhythm in SND was maintained when rhythmic PNA was eliminated during extreme hypocapnia. Fourth, during extreme hypercapnia, coherence of the rhythms in SND and PNA was drastically reduced. These results suggest that the slow rhythms in SND and PNA arise from separate oscillators that are normally coupled.