Activity in sympathetic neurons supplying skin and skeletal muscle in spinal cats

In anesthetized cats with intact neuraxis, vasoconstrictor neurons supplying skeletal muscle (MVC) and hairy and hairless skin (CVC), and sudomotor neurons innervating sweat glands (SM), exhibit distinct reflex patterns. MVC and SM are largely under excitatory, CVC under inhibitory control of various afferent input systems from the body surface and from the viscera. In chronic spinal animals all 3 types of sympathetic neurons exhibit some resting activity without cardiac and respiratory modulation. Sixty to 150 days after isolation of the neural circuits within the sympathetic systems within the spinal cord from their descending control systems by spinalization, these reflex patterns are very similar to those in animals with intact neuraxis. Important changes which do occur after spinalization are the following: CVC neurons are excited by stimulation of visceral afferents in spinal animals but inhibited in animals with intact neuraxis; noxious stimulation of skin leads to long-lasting after-effects in CVC and SM neurons in spinal animals. Comparison of reflexes among spinal animals and animals with intact neuraxis indicates that spinal circuits are probably important for the functioning of the sympathetic systems. It is possible that these circuits determine the typical reaction patterns seen in the sympathetic systems by integrating multisensory information from primary afferents and information from spinal descending fiber tracts.     

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)     

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.     

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.     

Functional properties of lumbar preganglionic neurones

Lumbar preganglionic neurones projecting through WRL₂ and L₃ to lumbar ganglia caudal to L₄ were investigated for those functional properties which are typical for postganglionic vasoconstrictor neurones supplying muscle and skin and for postganglionic sudomotor neurones. The properties tested were the cardiac rhythmicity of the activity and the reactions to systemic hypoxia, to noxious stimulation of skin and (in part of the experiments) to vibrational stimuli. Furthermore, resting activity and conduction velocities of the axons were measured.426 neurones were investigated. 311 (73%) of them were silent and could — as far as tested — not be excited by the afferent stimuli used. The conduction velocities of the axons of these neurones ranged from 0.5 to about 16 m/sec.115 neurones had resting activity of 0.1–4.6 impulses/sec. The conduction velocities of their axons ranged from 0.5 to about 12 m/sec. 80 preganglionic neurones with resting activity were classified on the basis of the reflexes in these neurones to afferent stimuli.Preganglionic neurones reacting like postganglionic vasoconstrictor neurones to muscle (excited by systemic hypoxia and/or by noxious stimulation of skin; with cardic rhythmicity) were classified as type 1 neurones (26 from 80 neurones tested). The resting activity of these neurones was1.8 ± 1.3impulses/sec(mean± 1S.D.). Their axons conducted with3.9 ± 2.2m/sec.Preganglionic neurones reacting like the majority of the postganglionic vasoconstrictor neurones to hairy and hairless skin (inhibited by systemic hypoxia and/or noxious cutaneous stimuli) were classified as type 2 neurones (48 from 80 neurones investigated). In 40% of these neurones the activity had cardiac rhythmicity. The resting activity was0.9 ± 0.6impulses/sec. The distribution of the conduction velocities of the axons of these neurones was bimodal. They conducted on the average with1.3 ± 0.6m/sec and6.6 ± 2.2m/sec respectively.A neurones were found (6 fronm 80 neurones) which were activated by vibrational stimuli (activation of Pacinian corpuscles by tapping on the hindfoot). Since this type of activation is typical for postganglionic sudomotor neurobes they were classified as type 3 neurones. The activity of these neurones had no cardiac rhythmicity.Indirect measurements of the conduction velocities of preganglionic axons converging onto postganglionic neurones supplying skeletal muscle and hairy skin yielded values which were statistically not different from the conduction velocities of the axons of type 1 and type 2 neurones respectively. These measurements support the classification into type 1 and type 2 preganglionic neurones. The implications of this study are discussed.     

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.     

Distribution of capsaicin-sensitive urinary bladder afferents in the rat spinal cord

The capsaicin-sensitive afferent innervation of the urinary bladder and the central nervous system distribution of urinary bladder afferents have been studied in the rat. Capsaicin-sensitive primary sensory neurones supplying the urinary bladder have been found in two groups of spinal ganglia located in the Th₁₃-L₂ and L₆-S₁ segments. Capsaicin-sensitive primary sensory afferents from the bladder terminate within Rexed's laminae I, V and X, and in the dorsal gray commissure of the lumbosacral spinal cord. In addition, the results point to a possible vagal sensory innervation of the urinary bladder.     

A gentle mechanical skin stimulation technique for inhibition of micturition contractions of the urinary bladder

Effects of gentle skin stimulation of various segmental areas on the micturition contractions of the urinary bladder were examined in anesthetized male rats. The bladder was expanded by infusing saline via urethral cannula until the bladder produced rhythmic micturition contractions as a consequence of rhythmic burst discharges of vesical pelvic efferent nerves. Gentle stimulation was applied for 1 min by slowly rolling on top of skin with an elastomer "roller". Rolling on the perineal area inhibited both micturition contractions and pelvic efferent discharges during and after stimulation. Stimulation of the hindlimb, abdomen and forelimb inhibited micturition contractions after stimulation ended, in this order of effectiveness. During stimulation of the perineal skin, the reflex increase in pelvic efferent discharges in response to bladder distension to a constant pressure was also inhibited up to 45% of its control response. The inhibition of the micturition contractions induced by perineal stimulation was abolished, to a large extent by the opioid receptor antagonist naloxone and completely by severing cutaneous nerves innervating the perineal skin. We recorded unitary afferent activity from cutaneous branches of the pudendal nerve and found that the fibers excited by stimulation were low-threshold mechanoreceptive Aβ, Aδ and C fibers. Discharge rates of afferent C fibers (7.9 Hz) were significantly higher than those of Aβ (2.2 Hz) and Aδ (2.9 Hz) afferents. The results suggest that low frequency excitation of low threshold cutaneous mechanoreceptive myelinated and unmyelinated fibers inhibits a vesico-pelvic parasympathetic reflex, mainly via release of opioids, leading to inhibition of micturition contraction.     

Identifying unique subtypes of spinal afferent nerve endings within the urinary bladder of mice

Spinal afferent neurons are responsible for the transduction and transmission of noxious (painful) stimuli and innocuous stimuli that do not reach conscious sensations from visceral organs to the central nervous system. Although the location of the nerve cell bodies of spinal afferents is well known to reside in dorsal root ganglia (DRG), the morphology and location of peripheral nerve endings of spinal afferents that transduce sensory stimuli into action potentials is poorly understood. The individual nerve endings of spinal afferents that innervate the urinary bladder have never been unequivocally identified in any species. We used an anterograde tracing technique developed in our laboratory to selectively label only spinal afferents. Mice were anesthetized and unilateral injections of dextran‐amine made into lumbosacral DRGs (L5‐S2). Seven to nine days postsurgery, mice were euthanized, the urinary bladder removed, then fresh‐fixed and stained for immunoreactivity to calcitonin‐gene‐related‐peptide (CGRP). Four distinct morphological types of spinal afferent ending in the bladder were identified. Three types existed in the detrusor muscle and one major type in the sub‐urothelium and urothelium. Most nerve endings were located in detrusor muscle where the three types could be identified as having: “branching”, “simple”, or “complex” morphology. The majority of spinal afferent nerve endings were CGRP‐immunoreactive. Single spinal afferent axons bifurcated many times upon entering the bladder and developed varicosities along their axon terminal endings. We present the first morphological identification of spinal afferent nerve endings in the mammalian urinary bladder.     

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.     

The effects of electrical stimulation of A and C visceral afferent fibres on the excitability of viscerosomatic neurones in the thoracic spinal cord of the cat

Single unit electrical activity has been recorded from viscerosomatic neurons in the lower thoracic spinal cord of decerebrate spinalized cats. The responses of the cells to electrical stimulation of afferent fibres in the splanchnic (SPLN) nerve and the effects of repetitive stimulation of somatic and visceral afferent C-fibres have been studied. Four groups of viscerosomatic neurones could be distinguished according to the type of visceral afferent input of the cells: (1) A-only cells (32.9%), driven only by stimulation of A delta afferent fibres in the SPLN nerve; (2) C-only cells (3%), driven only by stimulation of C afferent fibres in the SPLN nerve; (3) A + C cells (45.7%), driven by both A delta and C afferent fibres in the SPLN nerve; and (4) A + C? cells (18.6%), driven by A delta visceral afferents and showing signs of responsiveness to C-fibres though lacking a distinct response volley to visceral C-fibre activation. Two cells of the A + C group and located in lamina I of the dorsal horn responded to SPLN nerve stimulation in a manner consistent with the afferent fibre composition of the nerve, that is, showed evidence of strong monosynaptic links with SPLN afferent C-fibres and weaker responses to SPLN A delta afferents. Excitability changes of viscerosomatic neurones ('wind up', 'wind down' and changes in background activity) were also observed in the majority of neurones following electrical stimulation of somatic and of visceral afferent C-fibres.(ABSTRACT TRUNCATED AT 250 WORDS)     

N-methyl-D-aspartate antagonist applied to the spinal cord hindlimb enlargement reduces the amplitude of flexion reflex in the turtle

APV (D(-)-2-amino-5-phosphonovalerate), an NMDA (N-methyl-D-aspartate) antagonist, was applied in situ onto segments of the hindlimb enlargement of the turtle spinal cord. APV reduced the response amplitude of the flexion reflex. In contrast, APV did not alter the responsiveness of the rostral scratch reflex. Afferents for the flexion reflex enter the spinal cord via the dorsal roots of the middle segment of the hindlimb enlargement; afferents for the rostral scratch reflex enter the spinal cord via dorsal roots located anterior to the hindlimb enlargement. The results are consistent with the hypothesis that sensory interneuron NMDA receptors, synaptically activated either directly or indirectly by nearby cutaneous afferent axons, play a role in the spinal cord processing of cutaneous information.     

Role of unmyelinated fibers in electroacupuncture cardiovascular responses

The afferent fiber type responsible for the transmission of sensory neural traffic to the central nervous system during acupoint stimulation is uncertain. Several early studies evaluating compound action potentials have suggested that myelinated fibers contribute to the afferent input of the autonomic reflex adjustments during electroacupuncture (EA). Our more recent data, employing single unit recordings of somatic afferents, show that both myelinated and unmyelinated fibers are stimulated by EA, although more finely myelinated than unmyelinated fibers are activated by low frequency, low current stimulation. We hypothesized in this study that unmyelinated group VI fibers also contribute significantly to the inhibitory influence of EA on cardiovascular pressor responses. We found that neonatal capsaicin-treated rats depleted of substance P from primary afferents were insensitive to the inhibitory EA effect during gastric distention. Thus, EA at P5-P6 reduced gastric distention-induced pressor responses from 19+/-3 to 11+/-2 mmHg in eight untreated rats while capsaicin-treated rats (n=9) were unresponsive to EA. Substance P containing neurons in dorsal root ganglion cells at Ti-T5 were significantly decreased in the capsaicin-treated rats from 47+/-4 to 22+/-4 cells. Treated compared to untreated rats also demonstrated a significantly (P<0.03) reduced number of group IV fibers identified with single unit recording techniques. This study demonstrates that the inhibitory effect of EA at P5-P6 on cardiovascular autonomic excitatory reflexes involves unmyelinated group IV fibers of the median nerves.     

Influence of the pattern of jejunal distension on mesenteric afferent sensitivity in the anaesthetized rat.

Vagal, spinal and intestino-fugal fibres all potentially transmit mechanosensory afferent information from the gastrointestinal tract. We aimed to characterize the relative mechanosensitivity of these three different afferent populations supplying the rat jejunum. Afferent nerve discharge was recorded from pentobarbitone-anaesthetized rats during different distension protocols. Saline ramp distension (1 mL min(-1)) and barostat ramp distension (2 mmHg 4 s(-1)) each evoked biphasic responses but with the latter significantly attenuated especially at low distending pressures. Barostat controlled phasic distensions (10-50 mmHg, 25 s) evoked an afferent response with a peak at the onset of distension adapting to a plateau level that was maintained and comparable to the barostat ramp responses at the corresponding pressures. Chronic subdiaphragmatic vagotomy significantly attenuated the low pressure component of the response to balloon ramp distension and both peak and plateau responses to phasic distension. Single unit analysis showed an absence of low threshold afferent activity after vagotomy while the response to fibres with wide-dynamic range and high threshold sensitivity were preserved hexamethonium had no effect on the responses to either ramp or phasic distension. These findings suggest that the nature of the distension stimulus is critical in determining the pattern of response observed from the various subpopulations of afferents supplying the bowel wall.     

Electro-acupuncture stimulation to a hindpaw and a hind leg produces different reflex responses in sympathoadrenal medullary function in anesthetized rats

The effects of electro-acupuncture stimulation (EAS) of two different areas of a hindlimb with different stimulus intensities on sympathoadrenal medullary functions were examined in anesthetized artificially ventilated rats. Two needles of 160 microm diameter and about 5 mm apart were inserted about 5 mm deep into a hindpaw (Chungyang, S42) or a hind leg (Tsusanli, S36) and current of various intensities passed to excite various afferent nerve fiber groups at a repetition rate of 20 Hz and pulse duration of 0.5 ms for 30-60 s. Fiber groups of afferent nerves stimulated in a hindlimb were monitored by recording evoked action potentials from the afferents innervating the areas stimulated. The sympathoadrenal medullary functions were monitored by recording adrenal sympathetic efferent nerve activity and secretion rates of catecholamines from the adrenal medulla. EAS of a hindpaw at a stimulus strength sufficient to excite the group III and IV somatic afferent fibers produced reflex increases in both adrenal sympathetic efferent nerve activity and the secretion rate of catecholamines. EAS of a hind leg at a stimulus strength sufficient to excite the group III and IV afferent fibers produced reflex responses of either increases or decreases in sympathoadrenal medullary functions. All responses of adrenal sympathetic efferent nerve activity were lost after cutting the afferent nerves ipsilateral to the stimulated areas, indicating that the responses are the reflexes whose afferents nerve pathway is composed of hindlimb somatic nerves. It is concluded that electro-acupuncture stimulation of a hindpaw causes an excitatory reflex, while that of a hind leg causes either excitatory or inhibitory reflex of sympathoadrenal medullary functions, even if both group III and IV somatic afferent fibers are stimulated.     

Primary afferent axons in the tract of Lissauer in the cat.

The present study is concerned with the numbers of primary afferent axons in the tract of Lissauer in the cat. The data show that approximately 27% of the axons in mid-thoracic and lumbosacral tracts are primary afferent fibers from the segment in question and another 20% of the axons are primary afferent fibers that come from nearby segments. In addition the data show that approximately 80% of the axons in the tract are unmyelinated and that there is a slightly higher proportion of unmyelinated as opposed to myelinated primary afferents. There is also a higher proportion of primary afferents in the medial as opposed to lateral parts of the tract, but there are significant numbers of primary afferents in lateral parts of the tract. Thus it seems clear that the tract contains more primary afferent fibers than was previously believed and if these data are confirmed, the conclusions will have a bearing on considerations of the primary afferent input into the dorsal horn.     

Substance P immunoreactive nerve terminals in the dorsolateral nucleus of the tractus solitarius: roles in the baroreceptor reflex

Physiological and light microscopic evidence suggest that substance P (SP) may be a neurotransmitter contained in first-order sensory baroreceptor afferents; however, ultrastructural support for this hypothesis is lacking. We have traced the central projections of the carotid sinus nerve (CSN) in the cat by utilizing the transganglionic transport of horseradish peroxidase (HRP). The dorsolateral subnucleus of the nucleus tractus solitarius (dlNTS) was processed for the histochemical visualization of transganglionically labeled CSN afferents and for the immunocytochemical visualization of SP by dual labeling light and electron microscopic methods. Either HRP or SP was readily identified in single-labeled unmyelinated axons, myelinated axons, and nerve terminals in the dlNTS. SP immunoreactivity was also identified in unmyelinated axons, myelinated axons, and nerve terminals in the dlNTS, which were simultaneously identified as CSN primary afferents. However, only 15% of CSN terminals in the dlNTS were immunoreactive for SP. Therefore, while the ultrastructural data support the hypothesis that SP immunoreactive first-order neurons are involved in the origination of the baroreceptor reflex, they suggest that only a modest part of the total sensory input conveyed from the carotid sinus baroreceptors to the dlNTS is mediated by SP immunoreactive CSN terminals. Five types of axo–axonic synapses were observed in the dlNTS. SP immunoreactive CSN afferents were very rarely involved in these synapses. Furthermore, SP terminals were never observed to form the presynaptic element in an axo–axonic synapse with a CSN afferent. Therefore, SP does not appear to be involved in the modulation of the baroreceptor reflex in the dlNTS.     

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.     

Supraspinal connections of neurones in the thoracic spinal cord of the cat: Ascending projections and effects of descending impulses

Single unit electrical activity has been recorded extracellularly from 103 neurones in the thoracic spinal cord of decerebrate cats. The responses of these neurons to electrical stimulation of cutaneous and visceral afferent fibres, their projection through ascending sensory pathways and the effects of descending impulses on the neurones have been studied. Of the 103 neurones recorded, 45 (43.7%) responded only to activation of cutaneous afferent fibres (‘Somatic’ neurones). Their recording sites were located mainly in laminae II, III and IV of the dorsal horn. The remaining 58 neurones (56.3%) responded to stimulation of cutaneous and visceral afferent fibres (‘Viscero-somatic’ neurones). Their recording sites were located in laminae I, V, VII and VIII of the grey matter. Sixteen neurones had axons projecting through ascending pathways: 6 were post-synaptic dorsal column cells (PSDC), 2 were spino-cervical tract cells (SCT), 5 projected through the contralateral ventro-lateral funiculus (VLQ) and 3 through the ipsilateral dorso-lateral funiculus (DLF). All PSDC cells were somatic and all VLQ neurones were viscero-somatic. Reversible spinalization of the animals by cold block resulted in a selective increase of the responses of viscero-somatic neurones to cutaneous and visceral C-fibre input. In some viscero-somatic neurones, cold block induced a reduction or abolition of the visceral input suggesting its mediation via supraspinal loops. Electrical stimulation of the ipsilateral DLF evoked non-specific inhibitions of all inputs to viscero-somatic neurones. These results are discussed in relation with the mechanisms of visceral sensation.     

Electrophysiological analysis of the ascending and descending components of the micturition reflex pathway in the rat

Electrophysiological techniques were used to examine the organization of the spinobulbospinal micturition reflex pathway in the rat. Electrical stimulation of afferent axons in the pelvic nerve evoked a long latency (136 +/- 41 ms) response on bladder postganglionic nerves, whereas stimulation in the dorsal pontine tegmentum elicited shorter latency firing (72 +/- 25 ms) on these nerves. Transection of the pelvic nerve eliminated these responses. Firing on the bladder postganglionic nerves was evoked by stimulation in a relatively limited area of the pons within and close to the laterodorsal tegmental nucleus (LDT) and adjacent ventral periaqueductal gray. Stimulation at sites ventral to this excitatory area inhibited at latencies of 107 +/- 11 ms the asynchronous firing on the bladder postganglionic nerves elicited by bladder distension. Electrical stimulation of afferents in the pelvic nerve evoked short latency (13 +/- 3 ms) negative field potentials in the dorsal part of the periaqueductal gray as well as long latency (42 +/- 7 ms) field potentials in and adjacent to the LDT. The responses were not altered by neuromuscular blockade. Similar responses were elicited by stimulation of afferent axons in the bladder nerves. The sum of the latencies of the ascending and descending pathways between the LDT and the pelvic nerve (i.e. 72 ms plus 42 ms = 114 ms) is comparable although somewhat shorter (22 ms) than the latency of the entire micturition reflex. These results provide further evidence that the micturition reflex in the rat is mediated by a spinobulbospinal pathway which passes through the dorsal pontine tegmentum, and that neurons in the periaqueductal gray as well as the LDT may play as important role in the regulation of the micturition.