Small Myelinated Fibers Research Articles

HISTOMETRIC CHARACTERISTICS AND REGENERATIVE CAPACITY IN WOBBLER MOUSE MOTOR NEURON DISEASE

In wobbler mice, histometric features and the response to injury in the less affected hindlimb nerves were compared with the severely affected forelimb nerves which were studied previously. The frequency of vacuolated anterior horn cells was not different at the individual ages studied (3 and 6 weeks, 3 months), but with the three ages combined, the frequency in the two systems differed significantly, being 1.7 +/- 0.9% in the cervical and 0.6 +/- 0.7% in the lumbar anterior horns (P = 0.005). The number of large myelinated nerve fibres (greater than or equal to 6 microns in diameter) became progressively reduced in ventral roots as the disease advanced, particularly for the forelimb. The total number of myelinated axons was well preserved in the hindlimb ventral roots because of an increase in the number of small myelinated nerve fibres. The regenerative capacity of motor neurons after axotomy (tested by radiolabelling growth cones) was well preserved. In the hindlimb, the increased numbers of small myelinated fibres in ventral roots and the normal regenerative capacity in the sciatic nerve and its branches contrast with the severely affected forelimb system. Our studies suggest that cervical and lumbar anterior horn cells have different susceptibilities to the wobbler gene, producing vulnerability in one system and disease resistance in the other.

Effect of pancreas allografts on the ultrastructure of sciatic nerves in diabetic Rats

In a long-term study using cyclosporin-A (Cy-A) as immunosuppressant in a dose of 10 mg/kg/day, pancreas-duodenum was transplanted from Brown-Norway donors to alloxan-diabetic Lewis rats (n = 190). The pancreas transplants (PT) were performed after 1, 3, 6, 9, 12, and 15 months of diabetes. Recipient rats were sacrificed between 3 and 12 months of graft retainment. The mean axonal cross-sectional area and relative percentage of small, medium, and large myelinated fibers was evaluated. Also studied were unmyelinated fiber, ovoid body, and glycogen inclusion densities. Control rats consisted of non-diabetic rats (n = 36), similar rats receiving Cy-A (n = 42), diabetic rats (n = 103), and diabetic rats receiving Cy-A (n = 45). It was found that PT had a beneficial effect on the axonal cross-sectional area of myelinated nerves, the relative percentage of the various sizes of nerve fibers, and the ovoid body density, especially so in early diabetes. The effects in late diabetes were less spectacular. PT did not prove beneficial to the glycogen inclusion and unmyelinated fiber densities.

Sensations conducted by large and small myelinated afferent fibres are lost simultaneously under compression-ischaemia block.

Sensations conducted by large and small myelinated afferent fibres are lost simultaneously under compression-ischaemia block.

Small diameter myelinated afferents produce vasodilatation but not plasma extravasation in rat skin.

1. Antidromic stimulation of the rat saphenous nerve at intensities sufficient to excite small myelinated (A delta) fibres but not unmyelinated (C) fibres produced evidence of a transient increase in skin blood flow in the saphenous nerve territory. The magnitude and time course of the vasodilatation depended on the number and frequency of stimuli delivered to the nerve. 2. There was no evidence of an accompanying plasma extravasation. 3. The results suggest that A delta afferent fibres are involved in axon reflex/axon response reactions as well as unmyelinated (C) afferent fibres.

Innervation of the guinea pig trachea: A quantitative morphological study of intrinsic neurons and extrinsic nerves

The innervation of the guinea pig trachea was studied in wholemount preparations stained for acetylcholinesterase, catecholamines, and substance P immunoreactivity and by electron microscopy. The majority of parasympathetic and afferent nerve fibres arrive from the vagus via branches of the recurrent laryngeal nerves. The recurrent laryngeal nerves are composed of several fascicles comprising 600-700 small myelinated fibres (2-5 microns diameter) and about 1,000-2,000 unmyelinated fibres; both components exit from the nerve and project in fine branches to the trachea. A separate component of 200-250 large myelinated fibres (more than 5 microns diameter) runs the full length of the nerve and innervates the striated muscles of the larynx. The recurrent laryngeal nerves are slightly asymmetric in their origin, length, number, and composition of fibres, with the right nerve being shorter but with more numerous and thinner myelinated fibres. At the distal end of the recurrent nerve, a fine branch called the ramus anastomoticus connects it to the superior laryngeal nerve. In the tracheal plexus, there are on average 222 ganglion cells (range 166-327), distributed mostly in small ganglia of 12 or fewer neurons. The ganglionated plexus is situated entirely outside the tracheal wall, overlying the smooth muscle. Ligation experiments show that sympathetic nerve fibres reach the trachea with the recurrent nerves via anastomoses between the sympathetic chain and vagus nerves, or occasionally with recurrent nerves directly, the largest being at the level of the ansa subclavia. There are also perivascular sympathetic nerve plexuses. Substance P immunoreactive fibres enter the trachea from the vagus nerves and by pathways similar to those of sympathetic nerves. There are also paraganglion cells within the recurrent laryngeal nerve that contain catecholamines and are surrounded by substance P immunoreactive fibres. After cervical vagotomy, all the large myelinated fibres of the ipsilateral recurrent laryngeal nerve degenerate and so do all but 10 or 20 small myelinated fibres and all but a few unmyelinated fibres. Degenerating fibres are found within the entire tracheal plexus, indicating bilateral innervation. The small myelinated fibres that survive cervical vagotomy probably represent sympathetic or afferent nerves with their cell bodies located in sympathetic or dorsal root ganglia.

A fulminant case of Guillain-Barre syndrome: topographic and fibre size related analysis of demyelinating changes.

In a necropsy case of early fulminant Guillain-Barré syndrome, demyelinating changes were observed throughout the peripheral nervous system, most conspicuous in the spinal nerve roots. The central/peripheral nervous system transition regions and most proximal zones of the roots escaped damage. In some root areas there were widespread early changes in myelin sheaths in the absence of significant infiltrates of inflammatory cells. In the fibre size analytical study, small myelinated fibres were preferentially involved.

Carbonic anhydrase activity in the peripheral nervous system of rat: the enzyme as a marker for muscle afferents

The distribution of carbonic anhydrase (CA) activity was determined histochemically using Hansson's cobalt phosphate method in cross-sections of peripheral nerves from rats. As the studied nerves contain either efferent, proprioceptive or exteroceptive myelinated fibres, our survey particularly focused on the question whether CA-reactive nerve fibres are functionally related. Intense CA activity was detected in all large diameter (8–12 μm) muscle afferents. The amount of similarly reactive cutaneous afferents was negligibly low (3.6%). Efferent fibres displayed only weak CA activity, which was confined to the small myelinated fibres (3–6 μm). Moderate staining could be assessed in medium-sized (4–11 μm) proprioceptive fibres. The same reactivity occurred in a sizeable percentage (11.4%) of exteroceptive afferents. Their diameters ranged from 4 to 11 μm. These results indicated, that high enzyme activity is found predominantly in large-calibre proprioceptive afferents, which according to Hunt's classification were identified as group IA and IB fibres. Further confirmation for our observations was obtained by demonstrating high levels of enzyme activity in primary nerve endings of muscle spindles (IA fibres) and in axon terminals of Golgi tendon organs (IB fibres) constantly. Finally possible functions for neuronal CA are discussed with respect to its high activity in a functionally related subpopulation of nerve fibres.

Radial and tibial nerve pathology of two lactating ewes with “kangaroo gait”

Radial and tibial nerves of two ewes with clinical signs of chronic “kangaroo gait” were examined by qualitative and quantitative techniques and compared to the same nerves of a clinically normal ewe in late lactation. In affected ewes, there was extensive axonal degeneration of myelinated fibres in the radial nerve. Large and small myelinated fibres were affected equally and unmyelinated fibres were normal. Nerve fibre regeneration was present. In contrast, tibial nerve changes in the “kangaroo giat” ewes were minimal. The chronic nature of the radial nerve pathology was consistent with the clinical time course of “kangaroo gait”. Regeneration may account for gradual improvement with eventual recovery in most chronically affected ewes. An episode of bilateral severe compression of a proximal radial nerve site is proposed as an explanation for the neuropathy, although the specific mechanism of this trauma is not known.

The extent of small fibre sensory neuropathy in diabetics with plantar foot ulceration.

Thresholds for cutaneous warming and cooling stimuli were measured in 20 diabetics with neuropathic foot ulcers. All patients had a profound disturbance of sensory perception in the ulcerated foot with complete loss of perception of warming; thresholds for vibration and cooling were highly abnormal in all but two patients. Measurements of thermal threshold were made on both feet in 10 patients: warming was lost bilaterally in all, and cooling was bilaterally absent in six. There was no clear pattern of sensory loss in those diabetics with unilateral foot ulceration to suggest that sensory impairment was the determining factor for the development of a plantar ulcer. Measurements of thermal thresholds were made at additional sites in 13 patients and although the most marked abnormalities of sensation were always found in the feet, in some severe neuropaths, abnormal thresholds on the hand and even the face were demonstrated. Thresholds for warming were invariably more abnormal than thresholds for cooling. The diabetics with neuropathic ulceration in this study all had severe generalised peripheral nerve disease involving large myelinated as well as both small myelinated and unmyelinated sensory fibres. The quantitative evidence on the distribution of sensory loss for thermal sensations supports the hypothesis that the neuropathic process affecting the small myelinated and unmyelinated fibres is length dependent.

The auditory system in methyl mercurial intoxication: a neuropathological investigation on 14 autopsy cases in Niigata, Japan.

The auditory system in 14 autopsy cases of methyl mercurial intoxication in Niigata was examined neuropathologically. In addition to degeneration of the transverse temporal gyrus, there was also either significant loss of small myelinated fibers or decrease of large neurons in the cochlear nerve, ventral cochlear nucleus and inferior colliculus. The degree of decrease was higher in acute patients than in the chronic, and was not correlative to the severity of cerebrovascular sclerosis. The hearing impairment in methyl mercurial intoxication may be induced by the combined degeneration of the neurons or nerve fibers of these structures in both acute and chronic patients.

Axonal degeneration and regeneration in sensory roots in a genital herpes model.

In a mouse model of genital herpes simplex virus type 2 (HSV-2) infection, roots of the lower spinal cord were examined 5 days to 6 months after inoculation. Using immunoperoxidase methods on paraffin sections, viral antigen was found in sensory ganglia, their proximal roots and distal nerves on days 5 and 6 after infection. In Epon sections, most mice had focal sensory root abnormalities in lower thoracic, lumbar or sacral levels. At days 7 and 10, lesions showed chiefly nerve fiber degeneration, particularly of large myelinated fibers. At 2 weeks, lesions contained relatively large bundles of small unmyelinated fibers with immature axon-Schwann cell relationships. From 3 to 6 weeks, lesions again contained many more small unmyelinated fibers than normal but, in increasing proportions, axons in bundles were isolated from their neighbors by Schwann cell cytoplasm, and Schwann cells having 1:1 relationships with axons showed mesaxon or thin myelin sheath formation. At later times, the proportion of small unmyelinated axons decreased in parallel with increased numbers of small myelinated axons. By 6 months, affected roots showed a relative reduction in large myelinated fibers, increased proportions of small myelinated fibers and Schwann cell nuclei. Numbers of unmyelinated fibers were reduced relative to 3- to 6-week lesions. Axonal degeneration and regeneration appears to be the chief pathological change in sensory roots in this model. If regenerated fibers arise from latently infected neurons, then establishment of latency is not a relatively silent event, but is associated with major long-lasting, morphologically detectable effects.

The pathology of early leprous neuropathy

A qualitative and quantitative study was made of early changes in nerves from 10 patients with tuberculoid or lepromatous type of leprosy. Five nerve biopsies, taken from sites remote from skin lesions, were considered to be unaffected when examined by paraffin histology but showed abnormalities in semi-thin resin sections and by electron microscopy; 5 showed mild to moderate involvement by paraffin histology. Changes in ‘unaffected’ nerves in both types of leprosy included the presence of subperineurial oedema; occasional evidence of fibre regeneration, sometimes with atypical features; increased numbers of small myelinated fibres, possibly a consequence of axonal atrophy; a few thinly remyelinated fibres, probably due to secondary demyelination, and some loss of unmyelinated axons. In more affected nerves there was variable loss of axons, both myelinated and unmyelinated. Demyelination was not a conspicuous feature; there was evidence of axonal atrophy in some fibres. Similarities in some of the changes observed in tuberculoid and lepromatous types of leprosy suggest a common mechanism of nerve damage, at least in the early stages. The presence of abnormalities in nerves at a distance from skin lesions implies a more diffuse nerve involvement than might have been expected in both types of leprosy.

Synaptic organization of primary axons in trigeminal nucleus oralis.

This report examines the morphology and synaptic connections of small-diameter primary trigeminal axons that terminate in the border zone (BZ) and ventrolateral (VL) subdivisions of rat trigeminal nucleus oralis (Vo). Primary axons were made visible for light and electron microscopic analysis by utilizing the method of anterograde transport of horseradish peroxidase. BZ receives the terminal arborizations of two different populations of small-diameter primary axons. One of these arises from unmyelinated parent fibers and terminates in the dorsal one-half of BZ, while the other has small myelinated parent branches that arborize throughout the subdivision. Terminating within VL are the arbors of a second population of small myelinated primary axons. The endings of all three populations of primary axons lie in synaptic glomeruli. Endings in both subdivisions derived from small myelinated parent fibers lie centrally in glomeruli. Those in VL form axodendritic synapses on numerous dendritic shafts and spines, while endings in BZ glomeruli make at least one axodendritic synapse on one or two dendritic shafts. Endings of unmyelinated primary axons in BZ lie at the periphery of glomeruli where each forms a single axodendritic synapse on a central dendrite. It is at these asymmetrical axodendritic synapses that these three populations of primary axons are thought to transfer their inputs directly to the dendritic arbors of second-order BZ and VL neurons. Common to all three glomeruli is one or more small axonal endings filled with flattened synaptic vesicles that establish axoaxonic synapses on the primary ending as well as axodendritic synapses on the dendritic element(s) receiving primary input. In view of their symmetrical to intermediate synaptic contacts, these endings are thought to belong to axons derived from at least one source that can inhibit or diminish the firing rate of second-order BZ and VL neurons in response to primary input.

SELECTIVE DEPRESSION BY ALFENTANIL OF GROUP III AND IV SOMATOSYMPATHETIC REFLEXES IN THE DOG

The effect of alfentanil on sympathetic reflexes evoked by supramaximal electrical stimulation of the radial nerve has been observed in five dogs anaesthetized with alpha-chloralose, paralysed with suxamethonium and artificially ventilated. In five dogs during the infusion of alfentanil at a rate of 20 micrograms kg-1 min-1 the late long latency sympathetic response evoked by unmyelinated fibres (group IV, C) was abolished at a mean dose of 68.8 (SE 2.85) micrograms kg-1. The infusion rate was then increased to 200 micrograms kg-1 min-1 and the early, short latency response evoked by small myelinated fibres (group III, A delta) was eliminated at mean total dose of 809 (SE 72) micrograms kg-1. When the infusion was stopped the group III reflex returned within 1-5 min and recovery to approximately 50% of control for both reflexes occurred within 15-60 min in different preparations. Mean arterial pressure and mean heart rate decreased from 140 (6) mm Hg and 132 (11) beat min-1 to 100 (7) mm Hg and 70 (6) beat min-1, respectively, by the time the group IV response was eliminated; that is, after a mean infusion time of 3.4 min. Thereafter, there was no further cardiovascular depression. Within 3 min of the administration of naloxone 2 mg i.v., the sympathetic reflexes, arterial pressure and heart rate returned to within control values.

Glutamate and substance P coexist in primary afferent terminals in the superficial laminae of spinal cord.

By light microscopic immunocytochemistry it has been previously shown that approximately equal to 70% of the neurons in rat dorsal root ganglia are labeled with an antiserum for glutamate conjugated to hemocyanin; the smaller among these neurons are also positive for substance P. By using a postembedding ImmunoGold method and electron microscopy, it is shown here that synaptic terminals in the superficial laminae of the spinal cord of rats selectively stain for the same glutamate antiserum. Immunolabeling is in small dome-shaped and in large scalloped synaptic terminals. Scalloped terminals are of two types. One type consists of dark terminals with many agranular vesicles of different size and a few large granular vesicles; these are probably endings of unmyelinated and small myelinated primary afferent fibers. The other type consists of light terminals with small agranular vesicles homogeneous in size with neurofilaments and many mitochondria; these are probably endings of larger myelinated primary afferent fibers. By means of double-labeling electron microscopic immunocytochemistry with colloidal gold particles of two different sizes, it is also shown here that substance P is present in only the dark type of glutamate-labeled scalloped terminals. The primary afferent origin of the terminals labeled by the antisera for glutamate and for substance P is demonstrated by a triple-labeling strategy: immunocytochemistry for both antisera on sections from rats in which dorsal rhizotomy or dorsal root ganglion injection of horseradish peroxidase conjugated to wheat germ agglutinin was performed. It is proposed that glutamate is the neurotransmitter in primary afferents mediating input from different peripheral receptor classes, including nociceptors. Effects of glutamate and substance P on spinal dorsal horn neurons may result from co-release of these two mediators from the same dorsal root afferent terminal.

The conduction velocities of peripheral nerve fibres conveying sensations of warming and cooling.

With the current practice of measuring thresholds for warming and cooling separately, the question of the exact nature of afferents subserving these sensations assumes new importance. Experiments to measure reaction times to warming and cooling stimuli at two sites on the lower limb are described. The conduction velocity for each sensation was estimated from the conduction distance and conduction time in the limb. The estimated mean conduction velocity for warming was 0.5, SD 0.2 m/s and cooling 2.1, SD 0.8 m/s. These figures confirm that the sensation of warming is conveyed in unmyelinated and cooling in small myelinated peripheral nerve fibres.

Conduction studies in peripheral cat nerve using implanted electrodes: II. The effects of prolonged constriction on regeneration of crushed nerve fibers

Arrays of chronically implanted electrodes were used to examine the time course of elongation and maturation of peripheral nerve fibers in the cat after crush of the tibial nerve in the proximal calf. Regeneration after crush alone was compared with crush 5 mm proximal to a tight constriction of the nerve. Regeneration was monitored by the progression of excitability along the electrode arrays on the tibial and plantar nerves. The sensitivity was sufficient to record the averaged activity in single nerve fibers allowing detection of the earliest regeneration. The diameters of the fastest regenerating fibers were estimated from the conduction velocity proximal to the site of crush. Both after crush alone, and after crush constriction, small myelinated fibers regenerated in front of large fibers. The rate of elongation after crush alone was 3.2 mm/day, whereas it was slower (P less than 0.02) distal to crush + constriction (2.2 mm/day). In both lesions, the extrapolated delay to onset of regeneration was 8 days. In observations up to 300 days after crush, maturation was delayed or impaired by the constriction, and the compound nerve action potential had a smaller amplitude and a dispersed shape. Transverse sections of nerves after crush + constriction showed a diminished number of large and an increased number of small fibers compared with crush alone, possibly due to persistent branching of regenerated fibers. After both crush alone and crush + constriction, regenerated fibers had similar g ratios, suggesting that myelination developed fully in fibers of diminished diameters.

Clinical and morphometric analysis of biopsied biceps brachii muscles in adult-onset chronic proximal spinal muscular atrophy. With special reference to intramuscular nerves.

In a group of 16 cases with adult-onset chronic proximal spinal muscular atrophy, we performed a morphometric analysis of 88 intramuscular nerves in biopsied biceps brachii muscles and correlated this analysis with clinical and histochemical parameters. The total number of large and small myelinated fibres in all fascicles was reduced to 71%, 23% and 83% of control values, respectively. The percentage reduction of large myelinated fibres for each case was significantly correlated with duration of illness, biceps muscle power and histochemical atrophy factors. Histograms of large intramuscular nerve fascicles showed unimodal distributions and shifts to the left with single peak increases at 1 micron. The densities of small myelinated fibres in large fascicles correlated with counts of 'enclosed' muscle fibres in ATPase preparations. Compared with amyotrophic lateral sclerosis, a lesser reduction of large myelinated fibres, but a greater increase in small myelinated fibres, was noted in adult-onset chronic proximal spinal muscular atrophy. These findings imply that there is a less marked loss of myelinated nerve fibres with more effective reinnervation in adult-onset chronic proximal spinal muscular atrophy than in amyotrophic lateral sclerosis.

Inhibition of nociceptive responses of lumbar dorsal horn neurones by remote noxious afferent stimulation in the cat

In cats anaesthetized with nitrous oxide and sodium pentobarbital, multireceptive lumbar dorsal horn neurones excited by controlled noxious radiant heating of glabrous hind paw skin were recorded by extracellular microelectrodes. These noxious heat responses were inhibited by concomitant noxious stimulation of the ipsilateral forepaw or pinna, or repetitive electrical stimulation of the ipsilateral forelimb deep radial nerve. Similar extents of inhibition were produced by noxious peripheral stimulation and by deep radial nerve stimulation in repetitive trains at intensities sufficient to excite small myelinated fibres or unmyelinated fibres. A greater inhibitory effect was produced by continuous repetitive high-intensity stimulation of the deep radial nerve. With a constant frequency (5 Hz) of continuous deep radial nerve stimulation, graded increases in stimulation intensity revealed the threshold for inhibition in the small myelinated fibre range, and an additional increment of the inhibitory effect when unmyelinated fibres were also activated. When suprathreshold for unmyelinated fibres, the efficacy of continuous deep radial nerve stimulation increased with graded increases in stimulation frequency, with a threshold frequency for inhibition between 0.5 and 1 Hz and maximal effect at 5 Hz. Two nociceptive-specific neurones studied were also inhibited by deep radial nerve stimulation. The results indicate that ‘diffuse noxious inhibitory controls’ (DNIC) occur in the cat and can be activated by remote electrical or natural noxious stimulation.

Spinal roots of rats poisoned with methylmercury: Physiology and pathology

The evoked potentials in the ventral and dorsal roots were recorded independently by stimulating the sciatic nerve of both control and methylmercury-poisoned rats. Poisoned rats showed markedly decreased amplitudes but normal latencies of the potentials evoked in the dorsal roots. Potentials evoked in the ventral roots had normal latencies and amplitudes. Pathological correlates indicated acute axonal degeneration of the dorsal roots, with a significant decrease of the large and small myelinated fiber densities. The ventral roots were histologically unremarkable. Our pathological confirmation of the electrophysiologic changes in the methylmercury-poisoned rats enables us to substantially assess the pathophysiological aspects of acute lesions in the spinal roots.