K. Dembowsky

Bulbospinal projections to the intermediolateral cell column; a neuroanatomical study

The location of those neurones in the brain stem that project to the intermediolateral column (ILC) from which preganglionic sympathetic neurones have their origin was studied by the method of retrograde transport of horseradish peroxidase (HRP). In cats 30--50 nl of a 30% HRP solution was injected into the region of the ILC at T3 or L1 on one side. After a survival period of 72 h the lower brain stem from C1 to the inferior colliculi was sectioned and prepared for histological study under brightfield illumination. Neurones stained with exogenous HRP were found in three regions: (a) in the ipsilateral, dorsomedial part of the nucleus of the solitary tract (NTS) (43% of all labeled neurones), in the cranial part of the NTS, and also on the contralateral side (7%); (b) in the ventrolateral reticular formation beginning at the level of the obex up to 8 mm cranial to the obex (25% ipsilateral, 3% contralateral); and (c) in the ventral part of the raphe nuclei (postpyramidal and inferior central nucleus) from 2 to 9 mm cranial to the obex (22%).

Afferent connections and spinal projections of the pressor region in the rostral ventrolateral medulla of the cat

Following microinjection of wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP) into the pressor region of the rostral ventrolateral medulla of the cat, the medulla, pons and hypothalamus were examined for retrogradely labelled cell bodies, while the thoracolumbar segments of the spinal cord were examined for anterogradely labelled axons. Dense groups of labelled cells were found in the following areas: (1) the nucleus of the solitary tract, particularly the medial, ventrolateral and commissural subnuclei; (2) the ambiguous complex and immediately surrounding area; (3) the Kölliker-Fuse nucleus in the pons; (4) the paraventricular nucleus and lateral hypothalamic area. In the spinal cord, labelled axons formed a band extending throughout the dorsolateral and ventrolateral funiculi at thoracic segments, while terminal labelling was observed in the intermediolateral nucleus and to a lesser extent the central autonomic area, but not in other parts of the grey matter. The findings are discussed in relation to the role of the rostral ventrolateral medulla in cardiovascular regulation, particularly the baroreceptor reflex.

An intracellular study of the synaptic input to sympathetic preganglionic neurones of the third thoracic segment of the cat

In chloralose anaesthetized, paralyzed and artificially ventilated cats intracellular recordings were obtained from sympathetic preganglionic neurones (SPN) of the third thoracic segment of the spinal cord identified by antidromic stimulation of the white ramus T3. The synaptic input to SPNs was assessed, in cats with intact neuraxis or spinalized at C3, by electrical stimulation of segmental afferent fibres in intercostal nerves and white rami of adjacent thoracic segments and by stimulation of the ipsi- and contralateral dorsolateral funiculus and of the dorsal root entry zone of the cervical spinal cord. In both preparations SPNs showed on-going synaptic activity which predominantly consisted of excitatory post-synaptic potentials (EPSPs). Inhibitory post-synaptic potentials (IPSPs) were rarely observed. EPSPs were single step (5 mV) or, less frequently, large (up to 20 mV) summation EPSPs. The proportion of SPNs showing very low levels of on-going activity was markedly higher in spinal than in intact cats. Stimulation of somatic and sympathetic afferent fibres evoked early EPSPs (amplitude 3 mV, latency 5-22.3 ms), and late, summation EPSPs (amplitude up to 20 mV, latency 27-55 ms). Early and late EPSPs were evoked in nearly all SPNs in which this synaptic input was tested in the intact preparation (from 79-93% of the SPNs). In spinal cats, early EPSPs were evoked in 88% of the SPNs, whereas late EPSPs were recorded only in half of the neurones. No evidence for a monosynaptic pathway from these segmental afferent fibres to SPNs was obtained. In both intact and spinal cats, stimulation of the dorsolateral funiculus evoked early and late EPSPs in SPNs. Late EPSPs were recorded in 70% and 37% of the SPNs in intact and spinal cats, respectively. Early EPSPs, however, were evoked in all neurones. The early EPSPs evoked by stimulation of the dorsolateral funiculus had several components which are suggested to arise from stimulation of descending excitatory pathways with different conduction velocities. The following conduction velocities were calculated in intact (spinal) cats: 9.5-25 m/s (7.8-13.2 m/s), 5.7-9.5 m/s (5.5-7.8 m/s), 3.8-5.7 m/s (3.2-5.5 m/s), and 2.6-3.8 m/s (2.1-3.2 m/s). EPSPs of these various groups were elicited in a varying percentage in SPNs. EPSPs of the most rapidly conducting pathway were subthreshold for the generation of action potentials; some EPSPs of this group had a constant latency suggesting a monosynaptic pathway to SPNs. Stimulation of the dorsal root entry zone at the cervical level yielded essentially the same results as stimulation of the dorsolateral funiculus.(ABSTRACT TRUNCATED AT 400 WORDS)

Neurones within the "chemosensitive area" on the ventral surface of the brainstem which project to the intermediolateral column.

SummaryWith the method of retrograde transport of horseradish peroxidase it has been demonstrated that neurones within the “chemosensitive area” of the brainstem project to the thoracic intermediolateral column. The function of these neurones is discussed in regard to the regulation of blood pressure.

Tonic descending inhibition of the spinal somato-sympathetic reflex from the lower brain stem

In chloralose-anaesthetized cats the spinal and supraspinal components of the somato-sympathetic reflex were evoked in the white ramus at T3 and/or L2 by stimulation of intercostal and spinal nerves. A reversible blockade of all ascending and descending spinal pathways was performed by cooling the spinal cord between the second and third cervical segment. Total blockade of conduction was produced at temperatures below 8.5 degrees C (281.5 K). The spinal blockade produced the following reversible effects. (1) Mean arterial pressure fell to 30-50 mm Hg (4.0-6.7 kpa) and the tonic background activity in the white ramus was reduced to 0-24% of control (mean 12.1 +/- 10.0%). (2) The amplitude of the early spinal reflex was increased from 100% to 111-316% (mean 200.9 +/- 49.5%, n = 49) at the thoracic level and to 125-342% (mean 181.4 +/- 74.4%, n = 7) at the lumbar level. The onset latency of the spinal reflex at T3 (range 8-21 msec) was shortened by 0.5-3.0 msec (mean 1.7 +/- 0.9 msec). (3) Supraspinal components were completely abolished. (4) Neither baroreceptor denervation nor midcollicular decerebration altered these effects. (5) The cold block induced increase of the amplitude of the spinal reflex was reduced by the alpha-adrenoceptor agonist clonidine; this effect was reversed by the alpha-adrenoceptor antagonist yohimbine. Selective cooling of the dorsolateral funiculus caused the same effects on the spinal and supraspinal reflexes as cold block of the whole spinal cord. From these findings it is concluded that in the anaesthetized cat the spinal component of the somato-sympathetic reflex is modulated by a descending tonic inhibition. This inhibition acts at both the thoracic and the lumbar level and its origin is in the medulla oblongata. This inhibition is, however, independent of baroreceptor inputs. The pathways descends in the dorsolateral funiculus. It is suggested that noradrenaline or adrenaline might be involved in the transmission of this inhibitory influence.

Three types of sympathetic preganglionic neurones with different electrophysiological properties are identified by intracellular recordings in the cat

AbstractIntracellular recordings were obtained from sympathetic preganglionic neurones (SPN) of the third thoracic segment in cats. Based on differences in their active and passive electrophysiological properties, three different types of SPNs were discerned:1)Type A neurones had a high resting membrane potential (RMP) (−60 to −86 mV) and a low input resistance (RN) 12–23 MΩ). Action potentials of these neurones had a pronounced IS-SD inflexion and a prominent shoulder in their falling phase. Spikes were rarely generated from the on-going synaptic activity.2)Type B neurones had a lower RMP (−48 to −65 mV) and a higher RN (21–37 MΩ). Their action potentials were characterized by an after-depolarization; they showed a slight IS-SD inflexion and a less pronounced shoulder in their falling phase. The after-depolarization was abolished by membrane hyperpolarization in a time dependent way. A hyperpolarization of at least 50 ms duration was required for its abolition. The after-depolarization was also abolished during repetitive discharges. In most of these neurones spikes were generated at irregular intervals and low rates (0.06–4.6 spikes/s) from the synaptic activity.3)Type C neurones were similar to type B neurones, but their action potentials did not show the after-depolarization. Additionally, spikes were generated at fairly regular intervals and rather high rates (0.8–6.5 spikes/s). The rate of spike repolarization of all neurones was markedly increased by hyperpolarization and decreased by membrane depolarization. Current-voltage curves of some type B and C neurones showed a marked rectification upon membrane hyperpolarization. In all type B and C neurones the presence of the A-current is indicated by the voltage trajectory at the end of hyperpolarizing current pulses. Current-frequency curves of type B and C neurones revealed discharge rates of SPNs as high as 100 spikes/s.

Tonic catecholaminergic inhibition of the spinal somato-sympathetic reflexes originating in the ventrolateral medulla oblongata.

In chloralose-anesthetized cats activity of the spinal and supraspinal components of the somato-sympathetic reflex were evoked in the white ramus at T3 by stimulation of the corresponding intercostal nerve. A blockade of all spinal pathways by means of a reversible cold blockade of the spinal cord at C2-C3 produced the following effects: (1) mean arterial blood pressure fell to 30-50 mm Hg and the tonic background activity in the white ramus was markedly reduced; (2) the amplitude of the spinal reflex was significantly increased and the supraspinal reflex was completely abolished; (3) localized cold block of the dorsolateral funiculus produced the same effect as cold block of the whole spinal cord; (4) neither baroreceptor denervation nor midcollicular decerebration altered these effects; and (5) the alpha-adrenoceptor agonist clonidine reduced the increased amplitude of the spinal reflex during cold blockade; this effect was reversed by the alpha-adrenoceptor antagonist yohimbine. Bilateral cold blockade of areas on the ventrolateral surface of the brain stem between the rootlets of the hypoglossal nerve and the trapezoid body caused the same effect on background and reflex activity in the white ramus as did spinal cord blockade. A mapping of the catecholaminergic (CA) neurons in the lower brain stem of the cat by means of the fluorescence method showed CA neurons in the ventrolateral medulla at two levels: (1) one group of neurons in the caudal medulla, which lies ventral and dorsal to the lateral reticular nucleus (corresponding to area A1 in the rat); and (2) a second group found more cranially and located ventrally to the facial nucleus (corresponding to area A5 in the rat). CA nerve terminals in the spinal cord mainly innervate the intermediolateral cell column. From these findings it is concluded that in the anesthetized cat the spinal component of the somato-sympathetic reflex is modulated by a descending tonic inhibition. This inhibition is independent of baroreceptor input. The pathways descend in the dorsolateral funiculus of the spinal cord, and it is suggested that they originate either in the cranial part of area A1 and/or area A5.

Characteristics of sympathetic reflexes evoked by electrical stimulation of phrenic nerve afferents

In chloralose-anaesthetized cats, sympathetic reflex responses were recorded in left cardiac and renal nerve during stimulation of afferent fibres in the ipsilateral phrenic nerve. In cardiac nerve, a late reflex potential with a mean onset latency of 75.6 +/- 13.8 ms was regularly recorded which, in 20% of the experiments, was preceded by an early, very small reflex component (latency between 35 and 52 ms). In contrast, in renal nerve only a single reflex component after a mean latency of 122.1 +/- 13.1 ms was observed. Bilateral microinjections of the GABA-agonist muscimol into the rostral ventrolateral medulla oblongata resulted in a nearly complete abolition of sympathetic background activity and in an 88% reduction of the late reflex amplitude with only small effects on the latency of the evoked potentials. Under this condition, an early reflex component was never observed to appear. After subsequent high cervical spinalization, the residual small potentials which persisted after bilateral muscimol injections were completely abolished and in cardiac nerve an early reflex potential with a mean latency of 45 +/- 10 ms was observed in all but one experiment. The early reflex was therefore referred to as a spinal reflex component which, however, is suppressed in most animals with an intact neuraxis. In the renal nerve a spinal response was only observed in one experiment after spinalization. The results suggest that sympathetic reflexes evoked by stimulation of phrenic nerve afferent fibres possess similar spinal and supraspinal pathways as previously described for somato-sympathetic and viscero-sympathetic reflexes.(ABSTRACT TRUNCATED AT 250 WORDS)

Some properties of the sympathoinhibition from the caudal ventrolateral medulla oblongata in the cat.

Publisher Summary Neurons in the caudal ventrolateral medulla oblongata (CVLM) play an important role in the control of sympathetic activity. The excitation of these neurons by small injections of excitatory amino acids causes an inhibition of sympathetic activity and a fall in arterial blood pressure. This chapter presents a study that clarifies the question that whether the sympathoinhibition from CVLM is because of post-synaptic inhibition or the disfacilitation of sympathetic preganglionic neurons (SPNs) by first studying the spinal pathway of this inhibitory response. If its pathway in the spinal cord could be dissociated from the descending pathway of sympathoexcitatory neurons in rostral ventrolateral medulla (RVLM) that runs in the dorso- and ventrolateral funiculus of the spinal cord, this would be strong evidence against the disfacilitation of SPNs. Because more direct evidence for either disfacilitation or post-synaptic inhibition should be provided by intracellular recordings of SPNs, this question has also been addressed by recording SPNs intracellularly and studying the effects of glutamate injections into CVLM on their membrane potential. It has also been studied if noradrenaline acting on α 2 -adrenergic receptors is involved as a transmitter in this inhibition and if this sympathoinhibition involves an afferent projection from the CVLM to the hypothalamus or cerebellum.

Central action of α-adrenoceptor agents on the baroreceptor reflex

Abstract In chloralose-anaesthetized cats the effects of intravenous application of the α1- and α2-adrenoceptor agonistic and antagonistic agents methoxamine, prazosin, B-HT 933 and rauwolscine were tested on baroreceptor reflex, sympathetic background activity and blood pressure. Sympathetic activity was recorded from the renal nerve and the efficacy of the central transmission of the baroreceptor reflex was measured by the duration of the complete inhibition of renal nerve activity during electrical stimulation of the left carotid sinus nerve. All baroreceptors were denervated by sectioning both carotid sinus and vagal nerves. The α1-agonist methoxamine increased baroreceptor-induced sympathoinhibition, sympathetic background activity and blood pressure. The α1-antagonist prazosin had the opposite effects. The α2-agonist B-HT 933 was most effective in augmenting the inhibitory response in sympathetic activity to baroreceptor stimulation; sympathetic background activity and blood pressure were also decreased. At low doses (50 μg/kg) the α2-antagonist rauwolscine reduced the baroreceptor sympathetic reflex inhibition and increased sympathetic activity and blood pressure. The effect of B-HT 933 upon the baroreceptor reflex could be completely antagonized by rauwolscine. These findings demonstrate a very effective facilitation of the baroreceptor reflex transmission by stimulation of central α2-adrenoceptors. Through such humoral-neuronal interaction circulating catecholamines are likely to modulate cardiovascular control.