The afferent and sympathetic components of the lumbar spinal outflow to the colon and pelvic organs in the cat. II. The lumbar splanchnic nerves

The cell bodies of the lumbar sensory and sympathetic pre- and postganglionic neurons that project to the inferior mesenteric ganglion in the lumbar splanchnic nerves of the cat have been labeled retrogradely with horseradish peroxidase applied to the central end of their cut axons near the inferior mesenteric ganglion. The numbers, segmental distribution, location, and size of these labeled somata have been determined quantitatively. After all the lumbar splanchnic nerves on one side of an animal were labeled, most labeled cell bodies were situated ipsilaterally in dorsal root ganglia, ganglia of the lumbar sympathetic trunk, and spinal cord segments L2-L5, with the maximum numbers in L3 and L4. A few labeled somata lay contralaterally or rostral to L2. After labeling of only one lumbar splanchnic nerve, the majority of cell bodies were found in the labeled segment, but a few were also present up to three segments rostral or caudal. These variations could always be attributed to extraspinal connections usually via the lumbar sympathetic trunk. Cross-sectional areas of labeled afferent somata were small relative to those of the entire population of dorsal root ganglion cells. Preganglionic cell bodies were labeled in the intermediate gray matter extending from its lateral border ventrolaterally across to the central canal. Two regions of high density were observed: one laterally just medial to the edge of the white matter and the other lateral to the central canal. The dorsolateral group lay somewhat medial and caudal to the usual limits of the intermediolateral column. Labeled preganglionic neurons were on the average larger than the unlabeled cells in the inferior mesenteric ganglion, with the group lying medially being larger than those that were laterally positioned. From the data, it is estimated that about 4,600 afferent axons, about 4,600 preganglionic axons, and about 2,800 postganglionic axons travel in the lumbar splanchnic nerves to the inferior mesenteric ganglion of the cat.     

The afferent and sympathetic components of the lumbar spinal outflow to the colon and pelvic organs in the cat. I. The hypogastric nerve

The cell bodies of the lumbar sensory and sympathetic pre- and postganglionic neurons that project to the pelvic organs in the hypogastric nerve of the cat have been labeled retrogradely with horseradish peroxidase applied to the central end of their cut axons. The numbers, segmental distribution, location, and size of these labeled somata have been determined quantitatively. Afferent and preganglionic cell bodies were located bilaterally in dorsal root ganglia and spinal cord segments L3-L5, with the maximum numbers in L4. Very few cells lay rostral to L3. Afferent cell bodies were generally very small in cross-sectional area relative to the entire population in the dorsal root ganglia. Most of the preganglionic cell bodies lay clustered just medial to the region of the intermediolateral column and extended caudally well beyond its usual limit in the upper part of L4. These neurons were, on the average, larger than the cells of the intermediolateral column itself, with the largest cells lying in the most medial positions. Most of the post-ganglionic somata were in the ipsilateral distal lobe of the inferior mesenteric ganglion, while some (usually less than 10%) lay in accessory ganglia along the lumbar splanchnic nerves and in paravertebral ganglia L3-L5. Postganglionic somata in the inferior mesenteric ganglion were larger than both labeled and unlabeled ganglion cells in the paravertebral ganglia. From the data, it is estimated that about 1,300 afferent neurons, about 1,700 preganglionic neurons, and about 17,000 postganglionic neurons project in each hypogastric nerve in the cat.     

The cell bodies of origin of sympathetic and sensory axons in some skin and muscle nerves of the cat hindlimb

Cell bodies of sensory and sympathetic axons projecting to skin and skeletal muscle of the cat hindlimb have been labeled retrogradely with horseradish peroxidase (HRP) in order to study location, size, and numbers of the somata of these neurons. HRP was applied to the freshly transected axons of nerves supplying hairy skin (superficial peroneal, SP; sural, Su), hairy and hairless skin of the paw (medial plantar, MP), or skeletal muscle (gastrocnemius-soleus, GS). Serial sections of lumbosacral dorsal root and sympathetic ganglia were studied after standard histochemical processing. Additionally, the numbers of myelinated fibers in the same nerves were determined. All sensory somata and 99.4% of sympathetic cell bodies were located ipsilaterally. Sensory somata were commonly restricted to two adjacent dorsal root ganglia (usually L6–7 for SP, MP; L7-S1 for Su, GS). Although sympathetic somata were more widely distributed rostrocaudally, their maximum frequency always occurred in the segmental ganglia immediately rostral to the sensory outflows, i.e., corresponding to rami communicantes grisei. Dimensions of sympathetic somata varied little between populations projecting to different tissues and were unimodally distributed. The size distributions of sensory somata were characterized by a peak between 10 and 20 μm radius, similar to sympathetic somata, and a varying smaller number of cells ranging up to 60 μm radius. Each nerve had a characteristic distribution profile of afferent somata. A population of very small cells was only present in GS, while the largest sensory somata in GS and MP were bigger than those in SP and Su. Numerical analysis of the data disclosed the characteristic composition of both myelinated and unmyelinated fibers in each nerve studied.     

Ascending pathways from the spinal cord in the himé salmon (landlocked red salmon, Oncorhynchus nerka)

The ascending pathways from the spinal cord of the himé salmon were anterogradely labelled by the cobaltic lysine method, and their courses and terminations were examined. Following application of the cobaltic lysine to the cut end of the spinal cord or injection of the cobalt at the 10th to 15th spinal segment, labelled axons were traced from the spinal cord to various regions in the rhombencephalon and the mesencephalon. The axons ascending from the dorsal funiculus gave off many terminals as they ascended to the lower medulla. This terminal area may be homologous to the nucleus funiculi dorsalis of other vertebrates, although cytoarchitectural differentiation of the area is not evident. The dorsal funicular fibers also formed some terminals in the vagus and glossopharyngeal motor nuclei and the nucleus fasciculi solitarii. The axons arising from the anterolateral funiculus ascended as the common trunk of the lemniscus spinalis. The lemniscus spinalis fibers distributed many axon terminals on their way through the lower medulla, and most probably made synaptic contacts with the peripheral dendrites of the cells of origin of the reticulospinal pathways throughout its rostrocaudal extent. They also projected to all the medullary cranial nerve motor nuclei (V, VI, VII, IX, and X). The rostral continuation of the lemniscus spinalis fibers entered the lemniscus lateralis and gave a rostrocaudally elongated terminal area in the nucleus lateralis of the torus semicircularis.(ABSTRACT TRUNCATED AT 250 WORDS)     

Afferent and sympathetic neurons projecting into lumbar visceral nerves of the male rat.

The cell bodies of thoracolumbar sensory and sympathetic pre- and postganglionic neurons that project to the colon and pelvic organs of the male rat were labeled retrogradely with horseradish peroxidase (HRP) in order to study numbers, segmental distribution, and location of the somata of these neurons quantitatively. HRP was applied to one hypogastric nerve (HGN), to the lumbar colonic nerves (LCN) and to the intermesenteric nerve (IMN). In order to estimate the significance of the branching of one axon into both hypogastric nerves a double-labeling technique with fluorogold and HRP was used. About 2640 neurons project into the two HGN added together (800 afferent, 1320 pre-, and 520 postganglionic), 4650 neurons into the LCN (360 afferent, 0 pre- and 4290 postganglionic), and 5990 into the IMN (1500 afferent, 1250 pre-, and 3240 postganglionic). About 4190 sympathetic postganglionic prevertebral neurons innervate the colon and pelvic organs, 1900 are located in the inferior mesenteric ganglion and 2290 in ganglia of the IMN. Considering the efferent component, the HGN mainly are preganglionic and the LCN exclusively postganglionic nerves. Branching of one axon into both HGN is a rare event and quantitatively negligible (less than 3%). Afferent neurons of all three nerves were found in the dorsal root ganglia (DRG) T12-L2 with the maximum in L1 and L2. The distribution of afferent neurons projecting into the LCN is shifted slightly more rostrally compared to neurons projecting into the HGN. The IMN distribution is located in a position in between. Preganglionic neurons projecting into the IMN are located in the spinal cord segments T12-L3 with the maximum in L1 and L2.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of oxytocin and vasopressin on thoracic sympathetic preganglionic neurones in the cat

When applied by iontophoresis onto single sympathetic preganglionic neurones in the intermediolateral nucleus of segments T₁–T₃ in the cat, oxytocin and vasopressin each had an excitatory effect. This effect consisted of a prolonged (30–300 sec) after-discharge following termination of application. These results indicate that oxytocin and vasopressin each exert excitatory effects on sympathetic preganglionic neurones and support the possibility that they may be chemical mediators of synaptic transmission in the intermediolateral nucleus, perhaps in cardioacceleratory and/or pressor pathways descending from the paraventricular nucleus of the hypothalamus.     

The origin of postganglionic and sensory axons in the cervical sympathetic trunk of the cat: A horseradish peroxidase study

The composition of the cervical sympathetic trunk (CST) in the cat is still not completely understood. The present study investigates, by the horseradish peroxidase (HRP) method of tracing neuronal connections, the presence of postganglionic and sensory neurons projecting via the CST. Following sympathectomy at the midcervical level and the application of HRP crystals to the cut ends of the CST which had been isolated from the surrounding by a 1.5% solution of agar-agar, labelled neurons were seen in the superior cervical (SCG), stellate (SG), inferior vagal ganglia (IVG), and spinal ganglia C₈–T₈. The maximum number of labelled nuerons was 536 in the SCG, 460 in the SG, 180 in the IVG, and 104 in spinal ganglia C₈–T₈.     

Sympathetic and afferent somata projecting in hindlimb nerves and the anatomical organization of the lumbar sympathetic nervous system of the rat

The anatomy of the sympathetic pathways from the spinal cord to the lumbar sympathetic trunk and the inferior mesenteric ganglion was studied systematically in the rat. Details of the arrangements of white and gray rami communicantes, sympathetic trunk ganglia, the intermesenteric nerve, and the lumbar splanchnic nerves are summarized. A modified nomenclature for the segmental ganglia of the paravertebral sympathetic chain is proposed. Cell bodies of sensory and sympathetic axons projecting to the skin and skeletal muscle of the rat hindlimb were labeled retrogradely with horseradish peroxidase (HRP) in order to study numbers, segmental distribution, and location of the somata of these neurons quantitatively. HRP was applied to the nerves supplying skeletal muscle (gastrocnemius-soleus, GS), hairy skin (sural, SU; saphenous, SA) and to a mixed nerve (tibial, TI). All sensory somata and 96.4% of the sympathetic cell bodies were located ipsilaterally. Sensory somata were commonly restricted to two adjacent dorsal root ganglia (usually L3-4 for SA; L4-5 for GS, TI; L5-6 for SU). Although the sympathetic somata were more widely distributed rostrocaudally (four to six segments), their maximum was always located one or two segments more cranially than the sensory outflow, i.e., corresponding to the rami communicantes grisei. From the data, it is estimated that 420 sympathetic and 530 afferent neurons project into GS, 590 and 3,610 into SU, 920 and 3,750 into SA, and 1,070 and 5,760 into TI. These absolute neuron numbers are compared with electron microscopic fiber counts from the literature.     

Effects of glutamate and aspartate on sympathetic preganglionic neurons in the upper thoracic intermediolateral nucleus of the cat

Glutamate and aspartate excited all spontaneously active sympathetic preganglionic neurons (SPNs) tested in the intermediolateral nucleus of spinal segments T1-T3. Most silent neurons could be induced to discharge but the remainder showed only a decrease in antidromic spike amplitude. These effects were typically fast (on, off less than 1 s). D,L-Homocysteic acid also produced excitation; this effect was typically slower. Glutamate and aspartate were usually equipotent; 20% were differentially sensitive to aspartate, 10% to glutamate.     

Segmental distribution of peptide-like immunoreactivity in cell bodies of the thoracolumbar sympathetic nuclei of the cat

The distribution of leucine-enkephalin, methionine-enkephalin, neurotensin, somatostatin, substance P, oxytocin, vasopressin, and neurophysin II in cell bodies of sympathetic autonomic nuclei of the thoracolumbar (T-L) spinal cord was studied immunohistochemically in cats after intrathecal administration of colchicine. Neurons containing only enkephalin-, neurotensin-, somatostatin-, and substance P-like immunoreactivity (ENK, NT, SS, SP, respectively) were found in the intermediolateral nucleus pars principalis (IMLp) and pars funicularis (IMLf), the nucleus intercalatus (IC), and the central autonomic area (CA). The size, shape, location, and numbers of the peptide-positive neurons in the IMLp, IMLf, and IC suggested that they were sympathetic preganglionic neurons (SPN). This was confirmed by a combined retrograde tracing/immunohistochemical study showing that most of these neurons at the levels of the T-L cord known to provide preganglionic fibers to the stellate ganglion were SPN. On the other hand, the functional identification of the neurons in the CA is uncertain as neurons were not observed which were both retrogradely labelled and contained ENK, NT, SS, or SP. Immunoreactive neurons in each area were counted in ten sections from each segment from C8 to L4. In the IMLp, the SPN with ENK were greatest in number (up to 25) in segments T4-T7 and L2-L3. The maximum number of SPN containing NT was found in segments T4-T7 (45 neurons). Of the four peptides, neurons containing SS were found in the greatest number (up to 48 in segments T2-T6); neurons containing SP were found in the smallest number (15 or fewer per segment). Few SPN containing each of the four peptides were found in the IC; CA neurons with ENK and NT were also few in number. A comparison of the numbers of immunoreactive neurons in the IML with earlier estimates for the total numbers of SPN in the IML at each level showed that the proportions of IML neurons containing each of the four peptides were fairly consistent throughout the T-L cord, with some exceptions. These results suggest that the innervation of visceral organs is not obviously peptide-specific, although some organs may be innervated by a greater proportion of SPN containing one of these peptides. Finally, the presence of ENK, NT, SS, and SP in SPN suggests that these four peptides act as neurotransmitters in preganglionic pathways to sympathetic ganglia.     

Synaptic organisation of lumbar sympathetic ganglia of guinea pigs: Serial section ultrastructural analysis of dye-filled sympathetic final motor neurons

The authors serially sectioned seven dye-filled neuronal somata and more than 1.6 mm of their dendrites from the lumbar sympathetic ganglia of guinea pigs and examined them ultrastructurally to determine the distribution of preganglionic synaptic inputs to their dendrites and cell bodies. Most of the surface of the neurons was covered with Schwann cells. Apposing boutons were rare, with an average density of one axosomatic bouton per 125 μm² of somatic membrane and one axodendritic bouton per 25 μm of dendrite. Many dendritic segments that were more than 50 μm long completely lacked any apposing boutons. Although the average density of apposing boutons was low, local densities could be high, so that clusters of up to four adjacent boutons occurred on cell bodies and dendrites alike. The spatial arrangement of the apposing boutons for each of the cells examined here was not significantly different from a random distribution. Consequently, the number of apposing boutons observed for any neuron was simply proportional to the amount of neuronal surface sampled in the serial section run. About 50% of boutons directly apposing the neurons lacked any detectable presynaptic specialisations. When they were present, the presynaptic densities had a mean length of about 220 nm, with no difference between boutons that made axosomatic or axodendritic appositions. By applying these data to complete reconstructions of the dendritic trees of dye-filled sympathetic neurons at the light microscopic level, the authors estimated that few neurons in the lumbar sympathetic chain of guinea pigs would receive more than 200 synapses or apposing boutons and that many of them would receive less than 100 synapses. Up to 50% of these boutons would be predicted to make axosomatic contacts. These new observations provide a strong morphological framework for a better understanding of how sympathetic final motor neurons process their preganglionic synaptic inputs. J. Comp. Neurol. 402:285–302, 1998. © 1998 Wiley-Liss, Inc.     

Segmental distribution of corticotropin-releasing factor-like and vasoactive intestinal peptide-like immunoreactivities in presumptive sympathetic preganglionic neurons of the cat

The distribution of corticotropin-releasing factor (CRF), vasoactive intestinal peptide (VIP), and luteinizing hormone-releasing hormone (LH-RH) in cell bodies of sympathetic autonomic nuclei of the thoracolumbar spinal cord was studied immunohistochemically in cats after intrathecal administration of colchicine. Neurons containing CRF-like immunoreactivity (CRFir) and VIP-like immunoreactivity (VIPir), but not LH-RH-like immunoreactivity, were found in the intermediolateral nucleus pars principalis (IMLp) and pars funicularis (IMLf). On the basis of identification in previous studies and the size, shape, and location of the immunoreactive cells, it is suggested that the neurons are sympathetic preganglionic neurons. Most of the neurons with CRFir (85.5%) were found in the IMLp in segments T2-T7 and L2-L3 and the remaining 14.5% were found in the IMLf in segments T2-T5. The largest proportion of neurons with VIPir (93.7%) was found in the IMLp in segments T2, T4-T7, and T9-T13. Only 6.3% of the neurons containing VIPir were found in the IMLf in segments T2, T4, T5, and T10. These findings suggest that CRF and VIP may participate in peptide-specific pathways to peripheral organs.     

Reflex responses of single salivatory neurons to stimulation of trigeminal sensory branches in the cat

Responses of 71 single salivatory neurons, identified by antidromic spikes evoked by stimulation of the chorda tympani, were tested to stimulation of the ipsilateral infraorbital (IO), inferior alveolar (IA) and lingual nerves (LN) in the cat. Fifty-one neurons responded with spike potentials to stimulation of one or more of these nerves (responsive type, R), while the remaining 20 neurons did not respond to stimulation of any of them (non-responsive type, NR). Thirty-three R neurons activated by stimulation of all of the 3 trigeminal afferent branches, while 12 neurons responded with spikes to stimulation of only one branch, usually of LN. Reflex spike responses appeared with a latency of 5.6–14.6 ms to LN stimulation, 6.4–15.7 ms to IO stimulation and 6.0–26.0 ms to IA stimultion. Impulses of both Aβ and Aδ afferent fibres of the trigeminal nerve were found to be effective for activation of salivatory neurons.     

Lateralisation of projections from the rostral ventrolateral medulla to sympathetic preganglionic neurons in the rat

Spinally projecting sympathoexcitatory neurons in the rostral ventrolateral medulla (RVLM), synapse with sympathetic preganglionic neurons (SPN) and regulate the activity of sympathetic nerves that control the heart, blood pressure and the adrenal medulla (AM). However, the degree of lateralization of the bulbospinal projections to SPN innervating specific targets is poorly understood. Three approaches were used in this study. Anterograde tracer was iontophoresed into a pressor site in the RVLM (left or right) and retrograde tracer injected into the superior cervical ganglion (SCG, right) and the AM (left). Close appositions between anterogradely labelled axons and retrogradely labelled SCG- or AM-SPN were counted. Projections to the SCG were bilateral. Projections to the AM were markedly ipsilateral. In the second part, retrograde tracers were injected unilaterally into the region of the intermediolateral cell column at spinal segment T2 or T8 on one side and the number of labelled neurons in the RVLM counted. The results from each level of injection were similar showing that 63–64% of the neurons were ipsilateral. Responses to glutamate microinjection into the RVLM on adrenal nerve (left) and superior cervical nerve (left) activity were measured. The ratio of the nerve responses was the same even when different sides of the RVLM were injected. The anterograde data strongly suggest that the RVLM projections to AM-SPN are predominantly ipsilateral. Although other experimental approaches also attempted to investigate lateralization, the retrograde data target different and functionally heterogeneous pools of SPN that may mask the ipsilateral projection to the AM. Similarly, chemical stimulation of the RVLM will excite not only monosynaptic projections but also polysynaptic projections that may also mask the predominant ipsilateral monosynaptic projection to AM.     

Comparative localization of colorectal sensory afferent central projections in the mouse spinal cord dorsal horn and caudal medulla dorsal vagal complex

The distal colon and rectum (colorectum) are innervated by spinal and vagal afferent pathways. The central circuits into which vagal and spinal afferents relay colorectal nociceptive information remain to be comparatively assessed. To address this, regional colorectal retrograde tracing and colorectal distension (CRD)-evoked neuronal activation were used to compare the circuits within the dorsal vagal complex (DVC) and dorsal horn (thoracolumbar [TL] and lumbosacral [LS] spinal levels) into which vagal and spinal colorectal afferents project. Vagal afferent projections were observed in the nucleus tractus solitarius (NTS), area postrema (AP), and dorsal motor nucleus of the vagus (DMV), labeled from the rostral colorectum. In the NTS, projections were opposed to catecholamine and pontine parabrachial nuclei (PbN)-projecting neurons. Spinal afferent projections were labeled from rostral through to caudal aspects of the colorectum. In the dorsal horn, the number of neurons activated by CRD was linked to pressure intensity, unlike in the DVC. In the NTS, 13% ± 0.6% of CRD-activated neurons projected to the PbN. In the dorsal horn, at the TL spinal level, afferent input was associated with PbN-projecting neurons in lamina I (LI), with 63% ± 3.15% of CRD-activated neurons in LI projecting to the PbN. On the other hand, at the LS spinal level, only 18% ± 0.6% of CRD-activated neurons in LI projected to the PbN. The collective data identify differences in the central neuroanatomy that support the disparate roles of vagal and spinal afferent signaling in the facilitation and modulation of colorectal nociceptive responses.     

Sympathetic and sensory neurons projecting into the cervical sympathetic trunk in the chicken.

The cell bodies of the sensory and sympathetic pre- and postganglionic neurons projecting into the cervical sympathetic trunk were retrogradely labeled with horseradish peroxidase in the chicken. Preganglionic neurons were located in the spinal segments T1-T6 (maximum T2), postganglionic neurons in the paravertebral ganglia T1-T3 (maximum T1) and sensory neurons in the dorsal root ganglia T1-T4 (maximum T1). Labeled preganglionic neurons were widely distributed across the intermediate gray matter and lateral funiculus, but the majority of them were located in the intermediomedial area dorsolateral to the central canal. The short and long axis diameters of labeled preganglionic neurons in this area decreased caudally. From the data of the present study, it is estimated that about 4190 preganglionic, about 450 postganglionic and about 390 sensory neurons project into the cervical sympathetic trunk cranial to the paravertebral ganglion T1 in the chicken.     

Effect of preganglionic stimulation or chronic decentralization on neurotensin-like immunoreactivity in sympathetic ganglia of the cat

In pentobarbital-anesthetized cats, supramaximal stimulation (40 Hz, 2 h) of the preganglionic input to the acutely decentralized right stellate (RSG) or superior cervical (RSCG) ganglion resulted in a decrease in neurotensin (NT)-like immunoreactivity (IR), by 83% in the SG and by 46% in the SCG, as determined by radioimmunoassay. Chronic (7 days) decentralization of the ganglia resulted in a similar depletion of NT-like IR (SG: 86%; SCG: 76%). Supramaximal stimulation (40 Hz, 2 h) of the intact postganglionic outflow of either ganglion had no effect on NT-like IR. These data suggest that NT in the SG and SCG is present in preganglionic axons and is released by activation of these axons.     

Specific and potassium components in the depolarization of the Ia afferents in the spinal cord of the cat

In the cat spinal cord, primary afferent depolarization (PAD) of group Ia fibers of extensor muscles is produced by high-frequency stimulation (100 Hz) of group I muscle flexor afferents without significant increases in extracellular potassium. On the other hand, the PAD produced by stimulation of mixed and pure cutaneous nerves correlates well with increases in potassium ions. We conclude that the PAD produced by group I muscle afferents results from the activation of specific pathways making axo-axonic synapses with the Ia fiber terminals. The PAD of Ia fibers resulting from activation of cutaneous nerves involves instead unspecific accumulation of potassium ions.