Localization of sensory neurons traversing the stellate ganglion of the cat

The distribution of sensory cells whose axons traverse the stellate ganglion and project via sympathetic cardiac nerves to the heart of the cat has been examined quantitatively. Horseradish peroxidase (HRP) injected at multiple sites in the right stellate ganglion, or applied to the middle cardiac nerve, labelled small numbers of cells in the thoracic dorsal root ganglia (DRG) from T₁ to T₈. These cells were most numerous between T₂ and T₅ and were consistently small (< 40 μm) relative to other cells in the DRG. When HRP was applied to middle cardiac nerves, the numbers of labelled sensory cells always exceeded the numbers of myelinated axons counted in the same nerves from other cats. It is concluded that the distribution of the cells of cardiac sensory fibres is more extensive within thoracic DRG than has been previously reported, and it is suggested that such fibres travelling in the sympathetic cardiac nerves may be either myelinated or unmyelinated.     

The numbers of unmyelinated and myelinated axons in normal and regenerated rat saphenous nerves

Counts have been made of the numbers of unmyelinated and myelinated axons in the proximal and distal stumps of regenerated rat saphenous nerves and from equivalent sites in normal nerves. In the proximal part of normal nerves there were averages of 1 045 myelinated axons and 4 160 unmyelinated ones. Regenerated nerves contained the same number of myelinated axons in their proximal stumps but there was a 40% reduction in the unmyelinated axon count. In the distal stumps of these nerves the myelinated axon count had increased by an average of 620; this comes about because some regenerated myelinated axons support more than one process in the distal stump. In contrast, the number of unmyelinated axons was reduced further, from a mean of 2 476 in the proximal stump to one of 2 219.

The sizes of Schwann cell units in the normal and regenerated nerves were also noted. Schwann cell units in the proximal and distal stumps of the regenerated nerves were smaller than those in the normal ones.

These changes associated with unmyelinated axons in regenerated nerves are likely to contribute to the sensory, vasomotor and sudomotor abnormalities that sometimes occur after peripheral nerve injury and regeneration.     

Multipotentiality of Schwann cells in cross-anastomosed and grafted myelinated and unmyelinated nerves: quantitative microscopy and radioautography.

Cross-anastomoses and autogenous grafts of unmyelinated and myelinated nerves were examined by electron microscopy and radioautography to determine if Schwann cells are multipotential with regard to their capacity to produce myelin or to assume the configuration seen in unmyelinated fibres. Two groups of adult white mice were studied. (A) In one group, the myelinated phrenic nerve and the unmyelinated cervical sympathetic trunk (CST) were cross-anastomosed in the neck. From 2 to 6 months after anastomosis, previously unmyelinated distal stumps contained many myelinated fibres while phrenic nerves joined to proximal CSTs became largely unmyelinated. Radioautography of distal stumps indicated that proliferation of Schwann cells occurred mainly in the first few days after anastomosis but was also present to a similar extent in isolated stumps. (B) In other mice, CSTs were grafted to the myelinated sural nerves in the leg. One month later, the unmyelinated CSTs became myelinated and there was no radioautographic indication of Schwann cell migration from the sural nerve stump to the CST grafts. Thus, Schwann cell proliferation in distal stumps is an early local response independent of axonal influence. At later stages, axons from the proximal stumps cause indigenous Schwann cells in distal stumps from the previously unmyelinated nerves to produce myelin while Schwann cells from the previously unmyelinated nerves to produce myelin while Schwann cells from the previously myelinated nerves become associated with unmyelinated fibres. Consequently, the regenerated distal nerve resembled the proximal stump. It is suggested that this change is possible because Schwann cells which divide after nerve injury reacquire the developmental multipotentiality which permits them to respond to aoxonal influences.     

Spinal segmental preganglionic outflow to cervical sympathetic trunk and postganglionic cardiac sympathetic nerves

The aim of this study was to verify in which spinal cord segments the preganglionic neurones projecting to the cervical sympathetic trunk or converging onto the somata of the postganglionic cardiac sympathetic neurones are located in cats. The thoracic white rami T1 to T5 were electrically stimulated and the evoked responses were recorded in the cervical sympathetic trunks and postganglionic cardiac nerves. The responses were mostly evoked by electrical stimulation of group B preganglionic fibres. The maximum amplitude of evoked responses in the cervical sympathetic trunk was obtained when the T2 white ramus was stimulated and decreased gradually when followed by the stimulation of T1, T3, T4 and T5 white rami. In most cases the maximum amplitude of evoked responses in the left inferior cardiac nerve, right inferior cardiac nerve and left middle cardiac nerve was obtained when the T3 white ramus was stimulated. The size of the responses decreased when more cranial and caudal white rami were stimulated. It was found that the somata of the postganglionic neurones of the right and left inferior cardiac nerves were placed in the right and left stellate ganglion, respectively. Somata of the postganglionic neurones with axons in the left middle cardiac nerve were mainly located in the left middle cervical ganglion and some in the left stellate ganglion.     

Golgi-like immunoperoxidase staining of dopamine neurons in the reticular formation of the rat brainstem using antibody to tyrosine-hydroxylase

Cell bodies of sympathetic and sensory axons projecting via the superficial peroneal (SP) nerve supplying hairy skin of the distal hindlimb have been labeled retrogradely with horseradish peroxidase (HRP) on both sides of three cats in which the left SP nerve had been cut and ligated about 5 months previously. Three SP nerves from unoperated cats have also been studied. The location, size, and numbers of labeled somata have been determined from serial sections of lumbosacral dorsal root and sympathetic chain ganglia after standard histochemical processing. The numbers of myelinated fibers in each nerve have also been counted. The segmental distributions of both sympathetic and sensory cell bodies were identical bilaterally in each operated animal, but the number of labeled neurons was reduced on the lesioned side. There were only about 31% of sympathetic and about 51% of sensory somata relative to the numbers on the contralateral side. The average total number of neurons labeled from SP nerves in unoperated animals was about 8% higher than on the control side of operated animals. The average number of myelinated fibers in the neuromatized nerves was not reduced with respect to that in the contralateral nerve and both of these were not significantly different from the number in unoperated animals. The dimensions of samples of labeled sympathetic and sensory somata were reduced on the side with the neuroma, both in comparison with the contralateral side and with unlabeled neurons at the same levels. The mean cross-sectional area of profiles of sympathetic ganglion cells was 76% of the control; of sensory ganglion cells, 65% of the control. Assuming that HRP labeling was not impaired, we conclude that large numbers of neurons with unmyelinated axons had degenerated in the neuromatized cutaneous nerves.     

Avian sympathetic cardiac fibers and their cells of origin: anatomical and electrophysiological characteristics

The sympathetic cardiac innervation of the pigeon was investigated to describe certain anatomical and physiological properties of the cardiac nerve fibers and their postganglionic cells of origin. The compound action potential of the right cardiac nerve has two major components, one conducting at 2.0–5.6 m/sec with no chronotropic effect on the heart and the other conducting at 0.4–2.0 m/sec with a cardioacceleratory effect. Postganglionic neurons responding antidromically to cardiac nerve stimulation were then studied in ganglion 14 which contains most cells of origin of the cardiac fibers. These neurons have refractory periods of approximately 4 msec, following frequencies of< 4Hz, and axons conducting at 0.4–2.0 m/sec; this conduction velocity range corresponds to the slower compound action potential component. Electron microscopy of the cardiac nerve revealed unmyelinated fibers ranging in diameter from 0.2 to 1.2 μm and a population of myelinated fibers 1.0–3.6 μm in diameter. The unmyelinated fibers account for the slower compound action potential component and are largely postganglionic cardioaccelerator axons. The myelinated fibers account for the faster compound action potential component which has no chronotropic effect and is not reflected in postganglionic antidromic latencies; it is suggested that these myelinated fibers are cardiac sympathetic afferents. This study thus establishes electrophysiological criteria for identifying cardiac postganglionic neurons and describes the anatomical basis of these criteria.     

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.     

Spinal segmental sympathetic outflow to cervical sympathetic trunk, vertebral nerve, inferior cardiac nerve and sympathetic fibres in the thoracic vagus

The spinal segmental localization of preganglionic neurons which convey activity to the sympathetic nerves, i.e. vertebral nerve, right inferior cardiac nerve, sympathetic fibres in the thoracic vagus and cervical sympathetic trunk, was determined on the right side in chloralose anaesthetized cats. For that purpose the upper thoracic white rami were electrically stimulated with a single pulse, suprathreshold for B and C fibres, and the evoked responses were recorded in the sympathetic nerves. The relative preganglionic input from each segment of the spinal cord to the four sympathetic nerves was determined from the size of the evoked responses. It was found that each sympathetic nerve receives a maximum preganglionic input from one segment of the spinal cord (dominant segment) and that the preganglionic input gradually decreased from neighbouring segments. The spinal segmental preganglionic outflow to the cervical sympathetic trunk, thoracic vagus, right inferior cardiac nerve and vertebral nerve gradually shifted from the most rostral to the most caudal spinal cord segments. In some cases, a marked postganglionic component was found in the cervical sympathetic trunk. It was evoked by preganglionic input from the same spinal cord segments which transmitted activity to the vertebral nerve. These results indicate that there is a fixed relation between the spinal segmental localization of preganglionic neurons and the branch of the stellate ganglion receiving the input from these neurons.     

The components of the hypogastric nerve in male and female guinea pigs

A quantitative study has been made of the neural components of the hypogastric nerves of male and female guinea pigs using retrograde transport of horseradish peroxidase (HRP) to identify the population of neurones projecting in the nerve trunk, and electronmicroscopic analysis of the myelinated and unmyelinated axons present. Application of HRP to the transected axons of the hypogastric nerve labelled the cell bodies of sensory neurones in lumbar and sacral dorsal root ganglia, preganglionic neurones in the lumbar and sacral spinal cord, and postganglionic neurones in the inferior mesenteric ganglion and in the lumbar paravertebral chain; some ganglion cells of the pelvic plexus were also labelled. The number and distribution of each type of neurone with axons in the hypogastric nerve differed between the sexes: in particular, about twice as many preganglionic axons were present in the male as in the female.     

Unmyelinated axons in the ventral roots of the cat lumbosacral enlargement

The ventral roots L₇ and S₁ of the cat spinal cord were examined with the light and electron microscopes. Differences in the morphology of Schwann cells associated with large myelinated fibers and with small myelinated or unmyelinated fibers were observed. The blood vessels were largely encircled by pericytes. The most noteworthy finding was that 29% of the axons in these roots were unmyelinated. These unmyelinated axons were greatly reduced in number proximal but not distal to a ventral rhizotomy. Furthermore, they were reduced in number following dorsal root ganglionectomy, but not after dorsal rhizotomy, sympathectomy or peripheral nerve section. It is concluded that the ventral roots of the lumbosacral enlargement contain a large population of unmyelinated fibers originating from dorsal root ganglion cells.     

Hypoglycaemia causes degeneration of large myelinated nerve fibres in the vagus nerve of insulin-treated diabetic BB/Wor rats

The aim of this study was to find out whether dysglycaemia causes neuropathy in the vagus nerve of insulin-treated diabetic BB/Wor rats. Specimens were collected from the left vagus nerve proximal and distal to the level of recurrent laryngeal branch and from the recurrent branch itself in control rats and diabetic BB/Wor rats subjected to hyper- or hypoglycaemia. Myelinated and unmyelinated axons were counted and myelinated axon diameters were measured by electron microscopy. In controls, the vagus nerve proximal to the recurrent branch exhibited three regions in terms of fibre composition: part A was mainly composed of large myelinated axons, part B contained small myelinated and unmyelinated axons, and part C contained mainly unmyelinated axons. The distal level resembled part C at the proximal level and the recurrent branch resembled parts A and B. In hyperglycaemic rats, a normal picture was found at the proximal and distal levels of the vagus nerve and in the recurrent branch. In hypoglycaemic rats, signs of past and ongoing degeneration and regeneration of large myelinated axons were found at the proximal and distal levels and in the recurrent branch. We conclude that hypoglycaemia elicits degenerative alterations in large myelinated axons in the vagus and recurrent laryngeal nerves in diabetic BB/Wor rats. The absence of signs of neuropathy in unmyelinated and small myelinated axons suggests that the sensory and autonomic components of the nerve are less affected. In contrast, the hyperglycaemic rats examined here did not show obvious degenerative alterations.     

Regeneration of unmyelinated and myelinated sensory nerve fibres studied by a retrograde tracer method

Regeneration of myelinated and unmyelinated sensory nerve fibres after a crush lesion of the rat sciatic nerve was investigated by means of retrograde labelling. The advantage of this method is that the degree of regeneration is estimated on the basis of sensory somata rather than the number of axons. Axonal counts do not reflect the number of regenerated neurons because of axonal branching and because myelinated axons form unmyelinated sprouts. Two days to 10 weeks after crushing, the distal sural or peroneal nerves were cut and exposed to fluoro-dextran. Large and small dorsal root ganglion cells that had been labelled, i.e., that had regenerated axons towards or beyond the injection site, were counted in serial sections. Large and small neurons with presumably myelinated and unmyelinated axons, respectively, were classified by immunostaining for neurofilaments. The axonal growth rate was 3.7 mm/day with no obvious differences between myelinated and unmyelinated axons. This contrasted with previous claims of two to three times faster regeneration rates of unmyelinated as compared to myelinated fibres. The initial delay was 0.55 days. Fewer small neurons were labelled relative to large neurons after crush and regeneration than in controls, indicating that regeneration of small neurons was less complete than that of large ones. This contrasted with the fact that unmyelinated axons in the regenerated sural nerve after 74 days were only slightly reduced.     

The sympathetic and sensory components of the caudal lumbar sympathetic trunk in the cat

The cell bodies of the lumbar sensory and sympathetic pre- and postganglionic neurons that project in the caudal lumbar sympathetic trunk of the cat have been labeled retrogradely with horseradish peroxidase applied to the central end of their cut axons. The application was made just proximal to the segmental ganglion that sends its gray rami to the L7 spinal nerve, and so identified the sympathetic outflow concerned primarily with the vasculature of the hindlimb and tail. The numbers, segmental distribution, location, and size of the labeled somata have been determined quantitatively. Labeled cell bodies were found ipsilaterally, but the segmental distributions of the different cell types were not matched. Afferent cell bodies lay in dorsal root ganglia L1-L5 (maximum L4), preganglionic cell bodies in spinal segments T10-L5 (maximum L2/3), and postganglionic cell bodies in ganglia L2-L5 (maximum L5). Both numbers and dimensions of labeled dorsal root ganglion cells were variable between experiments (maximum about 1,000); the majority were small relative to the entire population of sensory neurons. Labeled preganglionic cell bodies were located right across the intermediate region of the spinal cord, extending from the lateral part of the dorsolateral funiculus to the central canal. The highest density of labeled neurons lay at the border between the white and gray matter (corresponding to the intermediolateral cell column) with smaller proportions medially in L1-L2, and laterally in caudal L4-L5. Medial preganglionic neurons were generally larger than those lying in lateral positions. From the data, it is estimated that about 650 afferent, about 4,500 preganglionic, and some 2,500 postganglionic neurons project in each lumbar sympathetic trunk distal to the ganglion L5 in the cat.     

Axonal branching following crush lesions of peripheral nerves of rat

Branching of myelinated and unmyelinated nerve fibers in normal and regenerating personal and soleus nerves was studied by light and electron microscopy. There were at most 2% more myelinated and 13% more unmyelinated axons in the distal as compared with the proximal nerve segments. Two to four weeks after a crush lesion the distal axons became 2-3 times more numerous; thereafter their number decreased. The number of axons in the proximal nerve segment did not change. The number of myelinated sprouts in most regenerated nerves equalled the number of myelinated fibers in the proximal nerve, while the number of unmyelinated axons after 12-19 weeks was 18-60% higher than normal. Branching was not restricted to the crush region. The results indicate that following a crush lesion all axons branch but only branches of unmyelinated fibers persist for a prolonged period of time. It is tentatively suggested that regenerating axons branch when searching for a target and that when contact is made with the target this prevents additional branching and eliminates redundant branches. Myelinated axons are guided by existing Schwann cells, whereas unmyelinated axons do not follow predetermined pathways; this may explain their greater tendency to form permanent branches.     

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.     

Stimulation of sciatic nerve regeneration in the adult rat by low-intensity electric current

Morphometric analysis was used to evaluate regeneration in transected sciatic nerves of adult rats constantly stimulated with low-intensity direct current. The ends of the cut nerve were separated by a distance of 5 mm and inserted into a Silastic tube. Histological and electron microscopy criteria were used to measure and evaluate the cross-sectional area and the structure of the bridge connecting the distal and proximal stumps. After 3 weeks of stimulation (10 microA DC with distal cathode) the stimulated animals showed a cross-sectional bridge area twice the size of nonstimulated controls. The number of myelinated and unmyelinated axons, and the vascular areas, were also larger in the experimental group. This is the first quantitative analysis of low-intensity direct current-enhanced peripheral nerve regeneration in adult mammals.     

An analysis of the axon populations in the nerves to the pelvic viscera in the rat

The present study uses selective surgical ablations combined with electron microscopic analyses to determine the number of axons in yarious catagories in rat hypogastric, pelvic, and pudendal nerves, these being the nerves to the pelvic viscera in this animal. Unmyelinated fibers predominate in all of these nerves. One of the most significant findings is that the pelvic nerve contains almost as many postganglionic sympathetic fibers as the hypogastric nerve. Previous investigators thought that the pelvic nerve supplied the parasympathetic inflow and the hypogastric nerve the sympathetic inflow to the pelvic viscera. The finding that there is a sizable sympathetic component in the pelvic nerve negates this idea, at least for the rat, and presumably calls for a reevaluation of the syndromes that arise from pelvic as opposed to hypogastric nerve section. Other findings of interest are (1) that there are unmyelinated efferent axons in the pudendal nerve, indicating that the pudendal is not a typical somatic nerve, (2) that the hypogastric nerve has a very small sensory component, and (3) that there are fibers surviving in the distal stumps of all these nerves, particularly the pelvic and hypogastric nerves.     

Unmyelinated primary afferent fibers in dorsal funiculi of cat sacral spinal cord

The present study tests the hypothesis that there are numerous unmyelinated primary afferent fibers in cat posterior funiculi. The animals have unilateral dorsal rhizotomies from L6 to Ca3. One week later the axons of both S2 dorsal funiculi are counted. The data indicate that there are approximately 22,500 myelinated and 8,500 unmyelinated axons on the unoperated side and 11,000 myelinated and 3,900 unmyelinated axons on the operated side. On this basis, we suggest that 51% of the myelinated and 54% of the unmyelinated axons in cat dorsal funiculi arise from dorsal root ganglion cells and thus are primary afferent axons. If this is correct, then 71% of the primary afferent axons in the cat dorsal funiculus are myelinated and 29% are unmyelinated. The function of this large group of previously unsuspected fine sensory axons remains to be determined.     

Properties of mechanoreceptor afferent fibres in left inferior cardiac nerve

Single afferent fibres with receptive field in the heart chambers and great vessels were dissected from the thoracic sympathetic chain. The fibres were identified by the electrical stimulation of the left inferior cardiac nerve, which was left in continuity with the heart. Fifty-six percent of the afferent fibres from the left inferior cardiac nerve were spontaneously active, the rest was silent. The spontaneously active fibres with receptive field in the aorta or left ventricle fired in close relation to the cardiac cycle as revealed by constructing post-R-wave time histogram of their activity. Fibres with cardiac rhythmicity were all myelinated. The second group of the spontaneously active fibres had their receptive field in the left atrium. The activity of these fibres was slightly or not at all correlated with cardiac cycle. They were both myelinated and unmyelinated fibres.