Secondary functional properties of lumbar visceral preganglionic neurons

Preganglionic visceral vasoconstrictor (VVC) neurons and motility-regulating (MR) neurons and other visceral preganglionic neurons, which project in the lumbar splanchnic nerves, were analyzed for their segmental distribution, the conduction velocity of their axons, ongoing activity and reflexes elicited by electrical stimulation of visceral afferents in white rami and of somatic afferents in spinal nerves. Identified preganglionic neurons and neurons without ongoing and reflex activity were distributed over segments L1-L5. VVC neurons were distributed over segments L1-L4 and MR neurons over segments L3-L5. VVC axons conducted at 2.8 +/- 2.5 m/s (mean +/- 1 S.D., n = 49), MR axons at 8.1 +/- 4.7 m/s (n = 131). The ongoing activity of VVC neurons was 1.6 +/- 0.7 imp/s (n = 46), that of MR neurons 0.8 +/- 0.7 imp/s (n = 91). There was no correlation between the conduction velocity of preganglionic axons and the rate of ongoing activity for VVC and MR neurons. (4) Electrical stimulation of visceral afferents in white rami and of somatic afferents in spinal nerves elicited short-latency (less than 50 ms) and long-latency (greater than 50 ms) reflexes in practically all VVC neurons, but preferentially short-latency reflexes in only 50 to 60% of the MR neurons. These results show that VVC and MR neurons are not only different in their reflex patterns, elicited by stimulation of visceral receptors and of arterial baro- and chemoreceptors, but also in the 4 properties analyzed in this paper.     

Functional characterization of preganglionic neurons projecting in the lumbar splanchnic nerves: neurons regulating motility

Lumbar preganglionic neurons, which projected in the lumbar splanchnic nerves and were probably involved in regulating motility of colon and pelvic organs (motility-regulating, MR neurons), were analyzed for their discharge patterns. The responses of the neurons to the following stimuli were tested: stimulation of arterial baro- and chemoreceptors and of afferents from the urinary bladder, colon, mucosal skin of the anus and perianal hairy skin. The following findings were made: a total of 131 preganglionic neurons were classified as MR neurons; these reacted to natural stimulation of at least one of the afferent inputs from the urinary bladder, colon and anal and perianal skin. The ongoing activity of these neurons did not correlate with the cardiac cycle or the cycle of the artificial ventilation. Most of them did not respond to an increase of blood pressure produced by i.v. injection of adrenaline or noradrenaline; some showed a weak depression or weak excitation which, in the time course, was untypical for visceral vasoconstrictor neurons. Stimulation of arterial chemoreceptors either did not influence MR neurons or produced only a secondary response owing to contraction of the urinary bladder. Ninety-seven preganglionic MR neurons could be subclassified: MR1 neurons were excited by distension and contraction of the urinary bladder and/or inhibited by distension and contraction of the colon (n = 61), a few were excited from both organs (n = 4); MR2 neurons were inhibited by distension and contraction of the urinary bladder and/or excited by distension and contraction of the colon (n = 32). Ninety-five out of 121 MR neurons (78.5%) were excited, 10 (8%) were inhibited and 16 (13%) not influenced by mechanical shearing stimuli applied to the mucosal skin of the anus. Most neurons which were excited by anal stimulation were not influenced by mechanical stimulation of the perianal (perigenital) skin. Twenty-eight per cent of the MR neurons (18 out of 64) were excited or inhibited upon stimulation of perianal skin. A few of these (7 out of 64 neurons, 11%) were involved in reflex responses which were different from those elicited from anal skin. At present no further consistent subclassification of MR1 and MR2 neurons appears possible on the basis of the excitatory and inhibitory anal and perianal reflexes. The results show that the population of visceral preganglionic neurons, which are probably involved in regulation of motility of colon and pelvic organs, is not homogeneous and probably consists of several subpopulations.     

Responses of lumbar vasoconstrictor neurons supplying different vascular beds to graded baroreceptor stimuli in the cat

Lumbar sympathetic vasoconstrictor neurons supplying skeletal muscle, hairy skin and pelvic organs were tested for their responses to carotid baroreceptor stimulation in chlorase-anaesthetized cats. Using single- and few-fibre recordings, the responses of the different types of vasoconstrictor neuron to graded steps of non-pulsatile pressure from 110 to 260 mmHg in a vascularly isolated carotid sinus were analyzed quantitatively during the first 10 s of stimulation. The activity in all postganglionic muscle vasoconstrictor (MVC) neurons, preganglionic visceral vasoconstrictor (VVC) neurons and one third of the postganglionic cutaneous vasoconstrictor (CVC₁) neurons was strongly depressed by maximal by baroreceptor stimulation. Moreover, quantitative analysis revealed no significant differences of the baroreceptor sensitivity of MVC and CVC₁ neurons as compared with VVC neurons at all levels of carotid sinus pressure. In contrast, two-thirds of the postganglionic cutaneous vasoconstrictor (CVC₂) neurons exhibited a significantly weaker barosensitivity. The functional implications are discussed.     

Discharge patterns of motility-regulating neurons projecting in the lumbar splanchnic nerves to visceral stimuli in spinal cats

Reflexes in visceral preganglionic motility-regulating (MR) neurons which project in the lumbar splanchnic nerves were investigated in acutely spinalized cats. Some neurons were analyzed before and after spinalization. The stimuli used were mechanical stimulation of mucosal skin of the anus and of perianal (perigenital) hairy skin, and distension and contraction of urinary bladder and colon. Most MR neurons exhibited a reflex pattern which consists of the following components: excitation upon bladder distension, inhibition or no effect upon colon distension and excitation (or, rarely, no effect) upon anal stimulation. This is the reflex pattern of MR1 neurons. Some neurons were excited by anal stimulation but not affected from the colon and urinary bladder. Some were inhibited by anal and perianal stimulation but otherwise exhibited the reflex patterns of the MR1 neurons. Analysis of the reflexes before and after spinalization showed that, in particular, inhibition elicited by anal, perianal and bladder stimulation was abolished; inhibition elicited from the colon was enhanced after spinalization. It is concluded that the reflexes elicited in preganglionic lumbar visceral neurons by the natural stimuli probably use spinal pathways, with the afferent input occurring at the sacral spinal cord. These spinal reflex pathways are probably controlled by descending inhibitory and excitatory spinal systems from the supraspinal neuraxis.     

Influence of carotid and aortic baroreceptors on neurosecretory neurons in supraoptic nuclei

(1) The effects of arterial baroreceptor stimulation on the activity of 'identified' neurosecretory cells in supraoptic nuclei (SON) of the hypothalamus were investigated in anesthetized cats. (2) Stimulation of baroreceptors by distension of an 'isolated' carotid sinus greatly inhibited SON activity. A linear relationship was found to exist between stimulus intensity (an increase in the sinus pressure) and the degree of inhibition of SON neuron activity. This inhibitory effect was abolished by section of the sinus nerve. (3) The inhibitory effect of baroreceptors on SON neuron activity was most pronounced in the first 5 sec during stimulation, and the effect became less when the stimulus lasted for a long period. At the end of stimulation there was a transient reversal of the response. (4) The excitation of aortic baroreceptors by occluding descending aorta strongly inhibited SON neuron activity. (5) Occlusion of carotid arteries augmented the SON neuron activity. Electrical stimulation of the sinus nerve caused an excitation or an inhibition of SON neurons, depending on the stimulus intensities. Apparently these stimuli produced different degrees of excitation in baro- and chemoreceptors. (6) Combination of chemo- and baroreceptor stimulations revealed that the excitatory effect exerted by chemoreceptor stimulation on SON neurons was reversed or blocked by baroreceptor activation, indicating the powerful inhibitory influence of baroreceptors.     

Reflex patterns in postganglionic vasoconstrictor neurons following chronic nerve lesions

Lesions of limb nerves in man may be associated with a variety of painful disorders with trophic changes described by the generic term 'reflex sympathetic dystrophy'. Our hypothesis is that pain and trophic changes are produced by an abnormal discharge pattern in postganglionic neurons supplying the limb (see refs. 3,24). In relation to this hypothesis, reflex patterns in postganglionic vasoconstrictor neurons supplying the skin (CVC) and the skeletal muscle (MVC) of the cat hindlimb were investigated at various times after a peripheral nerve lesion had been produced. These reflex patterns were compared with those in animals without nerve lesions (control preparations). The following lesions were made: cutting and ligating the superficial peroneal nerve (skin nerve) with subsequent neuroma formation, suturing the central stump of the superficial peroneal nerve to the peripheral stumps of muscle branches of the deep peroneal nerve, suturing the central stumps of muscle branches of the deep peroneal nerve to the peripheral stump of the superficial peroneal nerve, cutting and resuturing the superficial peroneal nerve, deafferentation of the whole hindlimb. The responses of vasoconstrictor neurons to stimulation of arterial chemoreceptors, arterial baroreceptors (cardiac rhythmicity of postganglionic activity) and cutaneous nociceptors were tested. In the animals with nerve lesions, the following groups of postganglionic vasoconstrictor neurons were analyzed: neurons projecting to the lesioned nerve, neurons projecting to hairy skin through an intact skin nerve (sural nerve) and neurons projecting to skeletal muscle through intact muscle nerves. In control preparations without nerve lesions, MVC neurons were excited by stimulation of arterial chemoreceptors and cutaneous nociceptors and inhibited by stimulation of arterial baroreceptors. Most CVC neurons were inhibited by stimulation of chemoreceptors and nociceptors and weakly inhibited by stimulation of baroreceptors. In animals with nerve lesions a and b, many CVC neurons in the lesioned nerves, as well as in the non-lesioned cutaneous nerve nearby, behaved in the same manner as MVC neurons. With respect to the control, this difference proved to be statistically significant. In preparations with lesions a, b and c, MVC neurons did not change their reflex patterns. After nerve lesions d and e, no major changes of reflex patterns were observed in CVC and MVC neurons. The inhibitory influence of arterial baroreceptors on CVC activity decreased in deafferented preparations (lesion e).(ABSTRACT TRUNCATED AT 400 WORDS)     

Reticular formation of the lower brainstem. A common system for cardiorespiratory and somatomotor functions: discharge patterns of neighboring neurons influenced by cardiovascular and respiratory afferents

Experiments were done in dogs with chloralose-urethane anesthesia. Long-lasting extracellular recordings were made from the medial parts of the reticular formation of the lower brainstem for up to 250 min. The study is based on reactions of 103 neurons. The activities of 2 or 3 neighbouring neurons recorded under identical conditions with one electrode or of neurons recorded with two electrodes at the same time could be changed regularly and synchronously by experimental changes of hemodynamic or ventilatory parameters. Action potentials were separated by amplitude discrimination. Rhythmic pulsatile modulations were proved to be present in 78% of all neurons by post-event-time histograms triggered by the R-wave of the ECG. In the 96 neurons tested 86% changed their activity when arterial pressure was raised by inflating a balloon in the abdominal aorta (79% decreased and 7% increased their activity). In post-event-time histograms triggered by the start of inspiration, 83% of the neurons showed modulations of their activity with respiratory rhythm. Experimental lung inflation decreased the activity in 75% of the tested neurons, while experimental lung deflation activated 47% of the tested neurons. Stimulation of arterial chemoreceptors activated 77% of the tested neurons. It was thus demonstrated that receptors in the cardiovascular and respiratory systems exert an influence on nearly all neurons from which recordings were made in that part of the reticular formation. Arterial baroreceptors and lung stretch receptors revealed a generalized depressing effect on the neuronal activity while chemoreceptors exert a generalized augmenting effect. At different times of recording these neurons did not always react to the same extent to comparable stimulations of afferents.     

Baroreceptor and chemoreceptor reflexes in postganglionic neurones supplying skeletal muscle and hairy skin

Postganglionic neurons supplying skeletal muscle and hairy skin of the cat's hindlimb were investigated for their reactions to stimulation of carotid baroreceptors and chemoreceptors in chloralose-anaesthetized, immobilized and artificially ventilated animals. The baroreceptors were stimulated by the pulsatile blood pressure and by pressure increases applied to an isolated carotid sinus (carotid blind sac) leaving only one intact buffer nerve. Chemoreceptors were stimulated by systemic hypoxia and by intracarotid bolus injections of CO2-saturated saline. The cardiac rhythmicity of activity in the postganglionic neurones (evaluated from the post-R-wave histograms) and the neurone reactions to intracarotid pressure increases were fairly well correlated, with a coefficient of r = 0.84. The time course and magnitude of inhibitory responses to intracarotid pressure increases were identical in some of the cutaneous postganglionic neurones and postganglionic neurones supplying skeletal muscle. The majority of the postganglionic neurones supplying skin, however, exhibited a weaker inhibitory response and a different time course in their activity during and after the sinus pressure increases. Muscle postganglionic neurones were excited and the majority of the cutaneous postganglionic neurones were inhibited by stimulations of arterial chemoreceptors produced by systemic hypoxia and by intracarotid injection of CO2-saturated saline. Small fractions of the post-ganglionic neurones supplying skin were excited by stimulation of arterial chemoreceptors. The activity of these cutaneous postganglionic neurones showed a strong cardiac rhythmicity. It is proposed that those postganglionic neurones supplying skin which are affected by stimulation of baro- and chemoreceptors in the same manner as are the postganglionic neurones supplying skeletal muscle may innervate cutaneous resistance (nutritional) vessels. It is likewise suggested that those cutaneous postganglionic neurones inhibited by stimulation of arterial chemoreceptors and under weak control by the arterial baroreceptors may innervate cutaneous capacitance and shunt vessels.     

Viscero-sympathetic reflex responses to mechanical stimulation of pelvic viscera in the cat.

Viscero-sympathetic reflex responses to mechanical stimulation of urinary bladder and colon were studied in cutaneous vasoconstrictor (CVC) neurones supplying hairy skin, in muscle vasoconstrictor (MVC) neurones supplying skeletal muscle and in sudomotor (SM) neurones supplying the sweat glands of the central paw pad of the cat hindlimb. The cats were anaesthetized, paralysed and artificially ventilated. The vasoconstrictor activity was recorded from the axons of the postganglionic fibres that were isolated in filaments from the respective peripheral hindlimb nerves. The activity in the sudomotor neurones was monitored by recording the fast skin potential changes occurring on the surface of the central paw pad. Afferents from the urinary bladder and from the colon were stimulated by isotonic distension and isovolumetric contraction of the organs. Most CVC neurones with ongoing activity were inhibited by these stimuli; only a few CVC neurones were excited. The MVC and SM neurones were generally excited by the visceral stimuli, yet the size of the evoked skin potential changes was variable. The reflex responses elicited in the sympathetic outflow to the cat hindlimb by stimulation of visceral afferents from the pelvic organs are uniform with respect to the different types of afferent input system but differentiated with respect to the efferent output systems. Graded stimulation of the visceral afferents from the urinary bladder by isotonic pressure steps elicited graded reflex responses in CVC (threshold less than 30 mmHg) and MVC neurones (threshold less than 20 mmHg) and a graded increase of the arterial blood pressure (threshold less than 20 mmHg). These graded reflex responses are closely related to the quantitative activation of sacral afferent neurones with thin myelinated axons innervating the urinary bladder that are also responsible for eliciting the micturition reflex, but not to the quantitative activation of sacral afferent neurones with unmyelinated axons. The latter have thresholds of 40-50 mmHg intravesical pressure at which the size of the vesico-sympathetic reflexes in the vasoconstrictor neurones was about 50% of maximal size. This does not exclude the fact that activation of unmyelinated vesical afferents contributes to the vesico-sympathetic reflexes.     

Differentiated cardiovascular afferent regulation of locus coeruleus neurons and sympathetic nerves

The activity of brain norepinephrine (NE) neurons in the locus coeruleus (LC) and peripheral sympathetic nerve activity (NE-SNA) in the splanchnic/renal nerve were recorded simultaneously during alterations of arterial blood pressure and circulating blood volume. Utilizing this experimental procedure we have previously found that both central and peripheral NE neurons are inhibited during blood pressure elevation. Furthermore, both neuronal systems were found to be inhibited during blood volume load, an effect apparently mediated by vagal afferents. In the present study both brain NE-LC activity and NE-SNA were increased during blood volume depletion. However, during prolonged hemorrhage the initial excitation of NE-SNA was followed by a marked inhibition. In contrast, the increase in NE-LC activity remained throughout the volume depletion period. The responses of central and peripheral NE neurons during hemorrhage were abolished in animals subjected to bilateral cervical vagotomy. Nitroprusside- or phenylephrine-induced blood pressure variations were associated with reciprocal changes in both central and peripheral NE neuronal activity. The NE-LC responses to blood pressure variations were abolished after bilateral vagotomy. NE-SNA responses, on the other hand, persisted after bilateral vagotomy. Our present and previous findings show that brain NE-LC neurons, similarly to peripheral NE neurons in the splanchnic/renal nerve, are regulated by tonically active cardiovascular afferents. Whereas peripheral NE-SNA is regulated by both arterial (high pressure) baroreceptors and cardiac volume (low pressure) receptors, the NE-LC neurons seem exclusively regulated by cardiac volume (low pressure) receptors.     

Transmission of impulses from pre- to postganglionic vasoconstrictor and sudomotor neurons

Transmission of impulses of pre- to postganglionic neurons supplying skeletal muscle and skin of the cat's hindlimb and tail was investigated. The objective of the study was to determine whether these postganglionic neurons can be influenced from the preganglionic side by non-nicotinic synaptic mechanisms in the lumbar sympathetic chain ganglia. The activity of the postganglionic neurons was recorded from their axons being isolated from peripheral skin and muscle nerves. (1) Vasoconstrictor neurons can be activated by muscarinic action of released acetylcholine and by a non-cholinergic synaptic mechanism. This type of non-nicotinic excitation of postganglionic vasoconstrictor neurons requires the activation of thin, probably unmyelinated preganglionic axons and considerable summation. Postganglionic sudomotor and pilomotor neurons cannot be activated in this way. (2) Ongoing activity in postganglionic vasoconstrictor neurons, but not in sudomotor neurons, can be enhanced for up to 60 min by brief trains of stimuli applied to the preganglionic site. Also this enhancement requires the activation of thin preganglionic axons. (3) Stimulation of thin preganglionic axons leads to an activation of muscle vasoconstrictor neurons via non-nicotinic synaptic mechanisms in the ganglia after complete block of nicotine transmission. (4) Postganglionic vasoconstrictor neurons and sudomotor neurons may be inhibited by a catecholaminergic autogenic mechanism in the ganglia. (5) The results indicate that integration may take place in the sympathetic chain ganglia by other than divergent and convergent processes. In this integration muscarinic actions of released acetylcholine and non-cholinergic synaptic mechanisms may be involved.     

Rapid phasic baroreceptor inhibition of the activity in sympathetic preganglionic neurones does not change throughout the respiratory cycle

We tested the hypothesis that the inhibitory influence of rapid phasic arterial baroreceptor stimulation on activity in sympathetic preganglionic neurones is weaker in inspiration than in expiration. Using neurophysiological techniques, 59 single preganglionic neurones with typical reflex pattern of muscle vasoconstrictor neurones that projected in the cervical sympathetic trunk were analysed. The inhibitory modulation of the ongoing activity in these neurones by the pulsatile activation of the arterial baroreceptors was determined by constructing post-R-wave histograms separately for both respiratory phases (as indicated by the discharge in the phrenic nerve). Quantitative measurements showed that the inhibition of the activity in the preganglionic neurones following phasic stimulation of arterial baroreceptors by the pulse pressure wave was not statistically different in both respiratory phases, even with increased respiratory drive.     

The role of the RVLM neurons in the viscero-sympathetic reflex: a mini review

Neurons in the rostral ventrolateral medulla (RVLM neurons) receive inputs from various sources, including baroreceptors, and then regulate activity of sympathetic preganglionic neurons. Though RVLM neurons are assumed to mediate the viscero-sympathetic reflex, it has not been clarified yet. Here we give a brief overview of the participation of RVLM neurons in the viscero-sympathetic reflex. We conclude that RVLM neurons show excitatory and inhibitory responses to stimulation of sympathetic afferents and mediate multi-phase reflex responses of the sympathetic nerve.     

Autonomic cardiovascular responses to hypercapnia in conscious rats: the roles of the chemo- and baroreceptors

The role of the autonomic nervous system, the central and peripheral chemoreceptors, and the arterial baroreceptors was examined in the cardiovascular response to hypercapnia in conscious rats chronically instrumented for the measurement of arterial blood pressure (ABP), heart rate (HR), and renal sympathetic nerve activity (RSNA). Rats were exposed to hypercapnia (6% CO2), and the cardiovascular and autonomic nervous responses in intact and carotid chemo- and/or aortic denervated rats were compared. In intact and carotid chemo-denervated rats, hypercapnia induced significant increases in mean ABP (MABP) and RSNA, and a significant decrease in HR. The HR decrease was reversed by atropine and eliminated by bilateral aortic denervation, which procedure, however, did not affect the MABP or RSNA response. Bilateral carotid chemo-denervation did not affect the baroreflex control of HR, although this control was attenuated by aortic denervation. Hypercapnia did not affect baroreflex sensitivity in intact rats. These results suggest that hypercapnia induces an increase in MABP due to an activation of sympathetic nervous system via central chemoreceptors and a decrease in HR due to a secondary reflex activation of the parasympathetic nervous system via arterial baroreceptors in response to the rise in ABP. In addition, carotid chemoreceptors do not play a major role in the overall cardiovascular response to hypercapnia in conscious rats. The mechanism responsible for the parasympatho-excitation may also involve CO2 induced aortic chemoreceptor simulation.     

Role of Barrington’s nucleus in the activation of rat locus coeruleus neurons by colonic distension

The locus coeruleus (LC)-noradrenergic system, which has been implicated in arousal and attention, is activated by visceral stimuli such as colon and bladder distension. Neurons of Barrington's nucleus (the pontine micturition center) have been identified which project to both the LC and preganglionic column of the lumbosacral spinal cord. Thus, Barrington's nucleus is positioned to coordinate brain noradrenergic activity with pelvic visceral functions. The aim of this study was to determine whether LC activation by colonic distension was mediated by projections from Barrington's nucleus to the LC in the rat. Lesions of Barrington's nucleus were performed unilaterally by local injection of ibotenic acid (microg/microl, 90 nl) 10 days prior to recording: (i) ipsilateral spontaneous LC discharge rate; (ii) LC responses to colonic distension; and (iii) LC responses to sciatic nerve stimulation. In some rats LC activation by hypotensive challenge was also examined. Lesions of Barrington's nucleus significantly reduced LC activation by colon distension from a magnitude of 26.6+/-6% increase in discharge rate (n=8) to 6.9+/-3% (n=6), while having no effect on basal LC discharge rate. In contrast, LC responses to sciatic nerve stimulation were not altered in rats with lesions of Barrington's nucleus and LC neurons were still activated by hypotensive challenge. These results support the hypothesis that Barrington's nucleus selectively relays input from pelvic visceral afferents to the LC. This may serve as a limb in a circuit designed to coordinate central and peripheral responses to pelvic visceral stimuli.     

Changes of reflexes in vasoconstrictor neurons supplying the cat hindlimb following chronic nerve lesions: a model for studying mechanisms of reflex sympathetic dystrophy?

The generic term 'reflex sympathetic dystrophy' describes a clinical syndrome which sometimes develops after traumata at the extremities with lesions of nerves or --more rarely--after other events. The syndrome consists of the following components: pain (hyperpathia, allodynia), trophic changes of skin and deep tissues, dysregulation of sweating and cutaneous blood flow of the extremity concerned. It is assumed that all symptoms are produced by abnormal sympathetic activity. Interruption of the sympathetic activity to the affected extremity abolishes most of the pain and may lead to remission of the trophic changes. The hypothesis is that the trauma with lesion of the primary afferent axons leads subsequently to an abnormal state of the primary afferent neurons and to distorted processing of information in the spinal cord. As a consequence of this abnormal central state the activity in the sympathetic (vasomotor and sudomotor) supply to the affected extremity is distorted. The results are pain, trophic changes and dysregulations of autonomic effector organs. In some yet unknown way a vicious circle between periphery and spinal cord is established (afferent leads to spinal cord leads to sympathetic leads to afferent). This hypothesis was the starting point for analysis of the reflex pattern in postganglionic vasoconstrictor neurons supplying the cat hindlimb after chronic nerve lesions performed in the same limb (cutting and ligating a skin nerve; suturing the central stump of a skin nerve to the peripheral stump of a muscle nerve). The results obtained show that the reciprocity of the reflex pattern which is normally observed between cutaneous and muscle vasoconstrictor neurons is lost in many animals. Cutaneous vasoconstrictor neurons are very similar to muscle vasoconstrictor neurons in their reactions to stimulation of arterial baroreceptors and chemoreceptors. If the same sequence of events also occurs in patients with reflex sympathetic dystrophy, it could explain the dysregulation of blood flow through skin and also the occurrence of trophic changes in the limb.     

Regulation of locus coeruleus neurons and splanchnic, sympathetic nerves by cardiovascular afferents

The brain norepinephrine (NE) neurons in the nucleus locus coeruleus (LC) have been claimed to be involved both in the regulation of behavioral functions, e.g. vigilance and arousal reactions, and in cardiovascular control. Recent studies from this laboratory have also shown that cardiovascular, vagal afferents can participate in the regulation of the LC neurons in the rat.

Utilizing electrophysiological techniques, we have now studied the effects of activation of blood volume receptors or arterial baroreceptors on the firing rate of single cells in the LC and, parallelly, on splanchnic, sympathetic discharge in the chloral hydrate anesthetized rat.

Blood volume load (0.5–5 ml heparinized blood, intravenously administered) induced a reduction in both LC neuronal firing rate and splanchnic nerve activity (SNA), effects which were readily and completely reversed by withdrawal of the corresponding amount of blood. In comparison, the central LC neurons were more sensitive to blood volume expansion than the peripheral splanchnic nerves. The effects of blood volume load on LC and SNA remained unaffected after deafferention of arterial baroreceptors.

Blood pressure elevation, induced by slow intravenous infusion of NE or angiotensin (AII) (total dose 2 μg/kg), caused an immediate reduction in both the firing rate of most of the LC cells tested as well as in SNA. While the effect on SNA was abolished by deafferentation of arterial baroreceptors, the effect on central LC activity remained largely unaffected.

Consequently, these data strengthen the concept that brain NE neurons in the LC are subject to control by peripheral blood volume receptors, analogously to peripheral sympathetic nerves. Arterial baroreceptors may still participate in the control of central noradrenergic nerve activity, but in contrast to their function for SNA they are not critical for the inhibition of LC neurons by blood pressure elevation. Rather, these two cardiovascular afferent systems may participate in the physiological regulation of the LC activity in a complimentary and convergent fashion.     

AMPA glutamatergic receptor-immunoreactive subunits are expressed in lumbosacral neurons of the spinal cord and neurons of the dorsal root and pelvic ganglia controlling pelvic functions in the rat

Sacral preganglionic neurons innervate the pelvic organs via a relay in the major pelvic ganglion. Pudendal motoneurons innervate striated muscles and sphincters of the lower urinary, genital and digestive tracts. The activity of these spinal neurons is regulated by sensory afferents of visceral and somatic origins. Glutamate is released by sensory afferents in the spinal cord, and interacts with a variety of receptor subtypes. The aim of the present study was to investigated the presence of AMPA glutamate receptor subunits (GluR1-GluR4) in the neural network controlling the lower urogenital and digestive tracts of male rats. We performed double-immunohistochemistry directed against a neuronal tracer, the cholera toxin beta subunit (Ctbeta) and each of the four receptor subunits. GluR1, GluR2 and GluR3 subunits were present in many sacral preganglionic neurons retrogradely labelled with Ctbeta applied to the pelvic nerve, and in some dorsolateral and dorsomedian motoneurons retrogradely labelled with Ctbeta injected in ischiocavernosus and bulbospongiosus muscles. The four subunits were detected in postganglionic neurons of the major pelvic ganglion retrogradely labelled with Ctbeta injected in the corpus cavernosum, and in some somata of sensory afferents of the L6 dorsal root ganglion labelled with Ctbeta applied to the dorsal penile nerve or injected in corpus cavernosum. The results provide a detailed knowledge of the neural targets expressing the various AMPA receptor subunits and suggest that part of the neural network that controls pelvic organs, including sensory afferents and postganglionic neurons, is sensitive to glutamate through the whole family of AMPA subunits.     

BDNF is a target-derived survival factor for arterial baroreceptor and chemoafferent primary sensory neurons.

Brain-derived neurotrophic factor (BDNF) supports survival of 50% of visceral afferent neurons in the nodose/petrosal sensory ganglion complex (NPG; Ernfors et al., 1994a; Jones et al., 1994; Conover et al., 1995; Liu et al., 1995; Erickson et al., 1996), including arterial chemoafferents that innervate the carotid body and are required for development of normal breathing (Erickson et al., 1996). However, the relationship between BDNF dependence of visceral afferents and the location and timing of BDNF expression in visceral tissues is unknown. The present study demonstrates that BDNF mRNA and protein are transiently expressed in NPG targets in the fetal cardiac outflow tract, including baroreceptor regions in the aortic arch, carotid sinus, and right subclavian artery, as well as in the carotid body. The period of BDNF expression corresponds to the onset of sensory innervation and to the time at which fetal NPG neurons are BDNF-dependent in vitro. Moreover, baroreceptor innervation is absent in newborn mice lacking BDNF. In addition to vascular targets, vascular afferents themselves express high levels of BDNF, both during and after the time they are BDNF-dependent. However, endogenous BDNF supports survival of fetal NPG neurons in vitro only under depolarizing conditions. Together, these data indicate two roles for BDNF during vascular afferent pathway development; initially, as a target-derived survival factor, and subsequently, as a signaling molecule produced by the afferents themselves. Furthermore, the fact that BDNF is required for survival of functionally distinct populations of vascular afferents demonstrates that trophic requirements of NPG neurons are not modality-specific but may instead be associated with innervation of particular organ systems.     

Intersegmental connections and interactions of myelinated somatic and visceral afferents with sympathetic preganglionic neurons in the unanesthetized spinal cat

The aim of this study was to obtain a measure of the interactions through exclusively spinal circuits, of myelinated afferents with sympathetic preganglionic neurons. Experiments were performed on 16 unanesthetized cats rendered insensitive by bilateral vertebral and carotid occlusion, whose spinal cords had been transected at C1 6-12 h before recording. The evoked responses of 68 tonically active sympathetic preganglionic neurons were recorded from filaments dissected from the cervical sympathetic trunk. Excitation, inhibition and excitation-inhibition sequences were evoked by electrical stimulation of radial, femoral and pelvic nerve afferents. Inhibition was most often observed during pelvic nerve stimulation. Ninety percent of the sympathetic preganglionic neurons tested responded to radial, 77% to femoral and 85% to pelvic nerve stimulation. These differences in percentage of units responding to the three nerves were statistically insignificant. Thus, in the acute spinal cat, the fraction of tonically active sympathetic preganglionic neurons whose activity can be influenced by myelinated afferents is independent of the length of the intraspinal pathway which conveys the input. A main difference between the long pathway (mediating the responses to femoral or pelvic nerve) and the short pathway (mediating the responses to radial nerve) seems to be the efficacy of their connections. While single shocks reliably evoked responses from the radial nerve, trains (200 Hz, 20 msec) were usually necessary to elicit responses from femoral or pelvic nerve, indicating a requirement for summation in the long pathway.