Fiber composition of the rat sciatic nerve

The rat sciatic nerve originates from the spinal segments L4-L6. It is unifascicular at the trochanter; 5-7 mm distally, the nerve splits into two and then into four fascicles. The tibial portion gives rise to the tibial and the sural nerves, and the peroneal portion gives rise to the peroneal nerve and a cutaneous branch that perforates the lateral hamstring muscles to innervate the proximolateral face of the calf. The number and type of the axons in these branches were determined in light and electron micrographs of normal nerves, and after de-efferentation or sympathectomy. Deafferentation was technically not feasible because spinal ganglia and ventral roots were supplied by the same vascular plexus. The tibial nerve contained 1,000 motor and 3,500 myelinated afferent axons, 3,700 sympathetic axons, and 5,400 unmyelinated afferent axons. The peroneal nerve contained 600 motor and 1,300 myelinated afferent axons, 1,100 sympathetic axons and 3,000 unmyelinated afferent axons. The sural nerve contained 1,100 myelinated and 2,800 unmyelinated afferent axons; in addition, there were 1,500 unmyelinated sympathetic axons. The cutaneous branch consisted of 400 myelinated and 1,800 unmyelinated afferent axons. Thus, the entire sciatic nerve at midthigh is composed of about 27,000 axons; 6% are myelinated motor axons, 23% and 48% are myelinated and unmyelinated sensory axons, respectively, and 23% are unmyelinated sympathetic axons. The techniques used did not demonstrate sympathetic axons in the cutaneous branch and did not reveal the few motor axons contained in the sural nerve.     

Substance P- and calcitonin gene-related peptide immunoreactivity in primary afferent neurons of the cat's knee joint.

The distribution of substance P and calcitonin gene-related peptide was determined in primary afferent neurons of the medial and posterior articular nerve of the cat's knee joint. Perikarya of articular afferents were visualized by retrograde labelling with the fluorescent dye Fast Blue which was applied at the transected end of the peripheral nerves. Substance P was found in about 17% of labelled medial articular afferents and in about 16% of labelled posterior articular afferents, respectively, whereas calcitonin gene-related peptide was present in about 35 and 32% of the medial and posterior articular nerve cells, respectively. Taking into account that these neuropeptides are known to be co-localized, probably not more than one-third of the joint afferents contain substance P and/or calcitonin gene-related peptide. Quantification of cell diameters revealed that substance P was found only in small- or intermediate-sized perikarya (less than 50 microns) indicating that this peptide is predominantly found in unmyelinated neurons. Calcitonin gene-related peptide was present mainly in small- and intermediate- but also in some large-sized neurons (greater than 50 microns) providing evidence that this peptide is found in unmyelinated and to a lesser extent in myelinated neurons. This is consistent with previous studies that show that substance P and calcitonin gene-related peptide are present primarily in unmyelinated and thinly myelinated primary afferents. When the portion of substance P-positive neurons of the medial articular nerve is compared to the number of articular afferents displaying a nociceptive function as determined in earlier electrophysiological studies, it can be calculated that at most 30% of the nociceptive-specific articular afferents contain this neuropeptide.(ABSTRACT TRUNCATED AT 250 WORDS)     

Anatomy of the rat knee joint and fibre composition of a major articular nerve

Several recent reports discuss the role of joint nerves in arthritis. Many of these are based on studies in the rat. The aim of this study is to examine the anatomy of the rat knee joint, in search for a primary articular nerve, and to analyze the fibre composition of that nerve. The results show that the structure of the joint differs in some respects from the human knee. At the upper end of the bony patella a cartilaginous patella extends proximally, forming the anterior wall of the suprapatellar bursa. Distinct collateral ligaments are integrated in the joint capsule. The extensor digitorum longus muscle bridges the knee joint, originating from the lateral femoral epicondyle. The well-developed menisci contain pyramid-shaped ossicles. The cruciate ligaments are arranged like in the human knee. A large posterior (PAN) and a small medial (MAN) articular nerve can be identified. The PAN is composed of some 400 axons, about 80% of which are unmyelinated. All myelinated fibres are sensory. They present a unimodal size spectrum with a size range of 1-8 microns, and a predominance of small fibres. Specific denervations indicate that about 1/3 of the unmyelinated axons represent afferents, and some 2/3 are sympathetic efferents. Interestingly, neonatal capsaicin treatment did not influence the number of unmyelinated PAN axons. The functional significance of the numerous unmyelinated sympathetic and sensory PAN axons in the normal knee joint remains to be elucidated.     

Fibers in monkey posterior articular nerves

The distribution of axons in the posterior articular nerves in rhesus monkey has been studied in terms of ratio of myelinated to unmyelinated fibers, and size of myelinated fibers. Conduction velocity measurements were also made. The nerves contained 2,000 to 2,200 unmyelinated fibers of 0.2 to 1.2 μm. diameter. They contained 399 to 478 myelinated fibers, varying in size from 1.4 to 12.3 μm. Thus 80 to 85% of fibers in these nerves were unmyelinated. The maximum conduction velocity of 70 to 80 meters per second corresponded to an initial small deflection, with most fibers conducting at slower rates. These results are compared to previous reports which severely underestimated the number of unmyelinated fibers because electron microscope counts were not utilized. It is suggested that pain fibers from the knee joint of monkey make up much of this large population of unmyelinated axons. An unknown number of post-ganglionic sympathetic fibers is also included in the unmyelinated fiber group.     

Number, distribution and neuropeptide content of rat knee joint afferents

Retrograde tracing with Fluoro-Gold was used to identify the complete population of knee joint afferents in the lumbar dorsal root ganglia of adult female Wistar rats. There was an average of 581±31 (mean± S.D. ) afferents supplying each joint. These were found distributed from L1 to L5 with the great majority localised in the L3 and L4 ganglia. Electron microscopy of the posterior articular nerve of the knee revealed an average of 103±15 (mean± S.D. ) myelinated and 513±39 unmyelinated axonal profiles. Since about 50–60% of the unmyelinated profiles would be expected to be sympathetic efferents, these numbers are consistent with the numbers of afferents found by Fluoro-Gold retrograde tracing and suggest that the posterior articular nerve contains about 50% of the total number of knee joint afferents in the rat. Immunohistochemistry revealed that an average of 10% of identified joint afferents expressed substance P-like immunoreactivity and that 33% expressed calcitonin gene-related peptide-like immunoreactivity.     

An electron microscopic analysis of the left phrenic nerve in the rat

In this electron microscopic study, the axonal categories in the left phrenic nerve at its entrance to the diaphragm have been determined. At a level 3 mm rostral to the diaphragm, the left phrenic nerve contains approximately 700 axons: 57% are myelinated and 43% are unmyelinated. The dorsal root ganglion cells give rise to 31% of the myelinated axons and the ventral root contributes 69%. Of the unmyelinated axons, the dorsal root ganglion cell contributes 59%, the cervical sympathetic chain 24%, and 17% course through the ventral roots. These ventral root unmyelinated axons are presumably preganglionic efferents since the proximal stump of the ventral root showed no decrease in unmyelinated axons after ventral rhizotomy.     

Dichotomizing somatic nerve fibers exist in rats but they are rare

Electrophysiological methods that resolve problems of current spread were used to reinvestigate recent claims that over 40% of myelinated afferent neurons in rats have dichotomizing axons in which branches are maintained in two separate hindlimb nerves. Out of a sample of about 6400 axons recorded in rat hindlimb nerves including 3641 myelinated afferents, 14 axons were found to have a branch in both the sciatic nerve and a second somatic nerve. Examples were found of dichotomizing myelinated afferents, myelinated efferents and unmyelinated afferents.     

Effects of a substance P analogue with antagonist properties ([D-Arg1, D-Trp7,9, Leu11]substance P) on spontaneous activity of the adrenal sympathetic nerve and its evoked reflex discharges in response to somatic afferent stimulation

The present study was designed to test the effect of a substance P (SP) antagonist, [D-Arg1, D-Trp7,9, Leu11]substance P (Spantide), on spontaneous efferent activity of adrenal sympathetic nerve and the two A- and C-sympathetic reflex components evoked in adrenal sympathetic nerve by stimulation of myelinated A- and unmyelinated C-afferent nerve fibers of a hindlimb nerve, respectively, in anesthetized rats. The spontaneous adrenal nerve activity and both of the A- and C-sympathetic reflex components were proportionally reduced by intrathecal application of the SP antagonist (0.038-0.76 nmol), although the same doses did not affect A- and C-somatic reflex components. These data suggest that the SP antagonist has a greater effect on SP-containing neurons related to the generation of spontaneous activity of sympathetic neurons and somato-sympathetic reflex pathways than on SP-containing primary somatic afferent nerves and somato-somatic reflex pathways.     

Shoulder region of the rat: Anatomy and fiber composition of some suprascapular nerve branches

Background: The pathophysiology of chronic supraspinatus tendinitis is not fully understood. This may be due to the scarcity of experimental studies on this issue. Methods: In search for a system suitable for experimental analysis, the present study describes the relevant gross anatomy of the rat shoulder region (dissection), and examines the fiber composition of relevant suprascapular nerve branches (electron microscopy, selective denervations). Results: The rat shoulder region is similar to the human shoulder in terms of gross anatomy. The average suprascapular nerve (SSC) is derived mainly from the spinal cord segment C5 and contains 3,435 axons, 74% of which are unmyelinated. The supraspinatus branch (SSP) contains 627 fibers. Of the SSP fibers, 52% are myelinated, including 32% motor and 20% sensory axons. Of the C-fibers in the SSP 16% are sympathetic efferents and 32% are sensory. Many of the latter disappear after neonatal capsaicin treatment. The SSC emits a subacromial articular branch (ART), with some 260 axons, about 90% of which are unmyelinated. The myelinated ART fibers are sensory, and of the unmyelinated ones about 24% are sympathetic efferents and 66% are afferents. The latter resist neonatal capsaicin treatment. Conclusions: In view of the anatomy of the supraspinatus muscle, of the subacromial space, and of relevant nerves, the rat shoulder should be appropriate for experimental studies on inflammatory conditions in the subacromial space. © 1994 Wiley-Liss, Inc.     

An electron microscope study of vagus nerve composition in the ferret

Summary The total number of axons in the cervical and abdominal vagus nerves of the ferret was counted. The ratio of myelinated to non-myelinated, and afferent to efferent axons was determined. The fibre diameter spectrum of myelinated axons was measured. The total number of axons in the ferret cervical vagus is similar to other mammals (approximately 28,000); the majority of axons are afferent (approx. 24,000) and also the majority of axons are nonmyelinated (approx, 27,000). The dorsal abdominal trunk is about twice the size of the ventral trunk although both trunks have the same number of efferent axons. The abdominal vagal trunks are over 90% afferent.     

Unmyelinated axons in a muscle nerve

Summary In continuation of earlier studies on the innervation of the sternomastoid muscle of the rat, a detailed morphometric analysis was performed on the unmyelinated axons of the nerve, in normal rats and after extensive cervical sympathectomy.In 4 normal rats an average of 314 myelinated and 319 unmyelinated axons were present. 42 days after surgery, the 3 animals showed Horner's syndrome and a highly significant 40% loss of unmyelinated axons. We therefore suggest that 40% of the C-fibers in this nerve are postganglionic sympathetic efferents and that the remaining 60% are type IV fibers, i.e., unmyelinated afferents. Our counts also indicate that part of the Remak bundles of the Schwann cells contain only sympathetic axons, whereas others contain mixed groups of sympathetic and afferent axons. Myelinated nerve fibers were not lost due to sympathectomy.Unexpectedly, the 3 animals analyzed 7–13 days after surgery showed Horner's syndrome but only a 16% loss of unmyelinated axons, which was not even statistically significant. Morphological signs of degeneration and sprouting did not provide any clue, but a possible explanation would be that a transitory sprouting of the remaining afterent C-fibers or Schwann cells occurred.     

人周围神经内乙酰胆碱酯酶活性与神经束性质的鉴别

本文用Karnovsky乙酰胆碱酯酶组织化学方法,系统地观察了成人周围神经中乙酰胆碱酯酶的分布。材料包括20侧上肢主要神经干及其分支、部分下肢神经的肌支和皮支、6侧C_7脊神经前后根。皮神经内,有髓纤维酶反应阴性;大量无髓纤维显示强阳性酶反应,呈群块状分布。肌支神经内58%的有髓纤维酶反应阳性,酶活性局限于轴索内,髓鞘不显酶活性;示强阳性酶反应的无髓纤维远较皮神经少。C_7脊神经前根87%有髓纤维酶反应阳性,与肌支内酶反应阳性有髓纤维相似;后根纤维酶反应阴性;灰交通支内酶反应强阳性,与皮支内阳性无髓纤维酶染色强度一致。本文结果表明,周围神经运动束和感觉束内,有髓及无髓纤维酶活性都有明显不同;明确提出,酶染色有髓纤维来源脊神经前根运动纤维,具有强酶活性的无髓纤维是交感神经节后纤维。     

The distribution of nodal sprouts in a paralysed or partly denervated mouse muscle

The right gluteus maximus muscles of young adult mice were paralysed with botulinum toxin for up to 21 days or partly denervated by spinal root section for up to 63 days; the intramuscular and extramuscular nerves were then examined in the electron microscope in thin sections of tissue fixed conventionally or stained with zinc iodide-osmium tetroxide. Contralateral muscles were also examined as controls. The distribution of nodal sprouts in the nerves of the paralysed or partly denervated muscles was determined by calculating the mean ratios of unmyelinated to myelinated axons in nerve profiles containing different numbers of myelinated axons (intact or degenerating) and comparing these with control ratios.In paralysed muscles there was a significantly higher proportion of nerve profiles containing one or two unmyelinated axons alongside a single myelinated axon. Nerve profiles containing two or more myelinated axons did not show any increase in the proportion of unmyelinated axons. Thus, there is probably nodal sprouting in paralysed muscles which is restricted to the most distal nodes of the intramuscular nerves.In muscles partly denervated for 8 days there were significant increases in the proportion of unmyelinated axons in nerve profiles which had contained up to 5–10 myelinated axons. After 21 days of partial denervation, similar increases may have occurred in the larger intramuscular nerve profiles and after 63 days there were large increases in the proportion of unmyelinated axons in the main intramuscular nerve branches and in the extramuscular nerve. Nodal sprouting in response to partial denervation is therefore localised initially to the smaller, more distal nerve branches; at later times, some sprouts probably grow slowly in a disto-proximal direction along denervated Schwann cell pathways.The existence of nodal sprouts in paralysed muscles and their restricted distribution in paralysed and partly denervated muscles suggest that the nodal sprouting stimulus is produced by the muscle and acts only at distal nodes.     

Afferent aortic nerve fibers with their pathways in cardiac sympathetic nerves.

The effect of mechanical and chemical stimulation on activity of afferent aortic nerve fibers with pathways in the cardiac sympathetic nerves has been examined. Action potentials were derived from the second or third thoracic communicating ramus of the left side of anesthetized dogs. Thirty myelinated and 19 unmyelinated fibers responded to tapping the ascending aorta, aortic arch, and descending aorta. Both groups of fibers also responded to a rise as well as to a fall in aortic pressure. Spontaneous discharge of myelinated fibers was related to aortic pressure pulse whereas that of unmyelinated fibers was related to respiration. Asphyxia caused excitation of unmyelinated fibers but not of myelinated fibers. Both groups of fibers responded to topical application of lactic acid. Mechanical and chemical stimulation of the aorta after vagotomy caused a rise in systemic blood pressure and extension of the limbs. The results indicate the existence of afferent aortic fibers in the cardiac sympathetic nerves that cause circulatory and somatic responses.     

Differential expression of tetrodotoxin-resistant sodium channels Nav1.8 and Nav1.9 in normal and inflamed rats

In an attempt to understand mechanisms underlying peripheral sensitization of primary afferent fibers, we investigated the presence of the tetrodotoxin-resistant Na+ channel subunits Nav1.8 (SNS) and Nav1.9 (SNS2) on axons in digital nerves of normal and inflamed rat hindpaws. In normal animals, 14.3% of the unmyelinated and 10.7% of the myelinated axons labeled for the Nav1.8 subunit. These percentages significantly increased in 48 h inflamed animals to 22.0% (1.5-fold increase) and 57.5% (6-fold increase) for unmyelinated and myelinated axons, respectively. In normal animals, Nav1.9 labeled 9.9% of the unmyelinated and 2.1% of the myelinated axons and following inflammation, the proportion of Nav1.9-labeled unmyelinated axons significantly decreased to 3.0% with no change in the proportion of labeled myelinated axons. These data indicate that Nav1.8 and Nav1.9 subunits are transported to the periphery in normal animals and are differentially regulated during inflammation. The massive increase in Nav1.8 expression in myelinated axons suggests that these may contribute to peripheral sensitization and inflammatory hyperalgesia.     

The articulo-cardiac sympathetic reflex in spinalized, anesthetized rats

Somatic afferent regulation of heart rate by noxious knee joint stimulation has been proven in anesthetized cats to be a reflex response whose reflex center is in the brain and whose efferent arc is a cardiac sympathetic nerve. In the present study we examined whether articular stimulation could influence heart rate by this efferent sympathetic pathway in spinalized rats. In central nervous system (CNS)-intact rats, noxious articular movement of either the knee or elbow joint resulted in an increase in cardiac sympathetic nerve activity and heart rate. However, although in acutely spinalized rats a noxious movement of the elbow joint resulted in a significant increase in cardiac sympathetic nerve activity and heart rate, a noxious movement of the knee joint had no such effect and resulted in only a marginal increase in heart rate. Because this marginal increase was abolished by adrenalectomy suggests that it was due to the release of adrenal catecholamines. In conclusion, the spinal cord appears to be capable of mediating, by way of cardiac sympathetic nerves, the propriospinally induced reflex increase in heart rate that follows noxious stimulation of the elbow joint, but not the knee joint.     

Fibre composition of the ventral roots L7 and S1 in the owl monkey (Aotus trivirgatus)

Summary The ventral roots L7 and S1 of the owl monkeyAotus trivirgatus, were examined by electron microscopy. On average, these roots contain 2950 and 1837 myelinated axons respectively. In both roots the myelinated axons have bimodal size distributions, but the S1 root contains more small myelinated axons. Both roots contain a substantial proportion of unmyelinated axon profiles (UAP). In the L7 root the proportion of UAP decreases as the spinal cord is approached, from 19% distally to 5% in the juxtamedullary rootlets. Unmyelinated and very small myelinated CNS-type axons have not been observed in the L7 transitional region. The average S1 root contains some 40% unmyelinated axons at all examined proximo-distal levels. Unmyelinated/ very small myelinated axons are easily found on the CNS side of the S1 transitional region, in direct relation to motoraxon bundles. Bundles of unmyelinated and small myelinated axons occur in the ventral pia mater of both segments. The unmyelinated axons in the L7 root of the owl monkey appear to be arranged like those in the feline L7 ventral root, possibly representing afferents. It is likely that most unmyelinated and small myelinated axons in the ventral root S1 are autonomic efferents.     

Carotid sinus nerve terminals which are tyrosine hydroxylase immunoreactive are found in the commissural nucleus of the tractus solitarius

Summary Tyrosine hydroxylase immunoreactive sensory neurons in the petrosal ganglion selectively innervate the carotid body via the carotid sinus nerve. Central projections of the carotid sinus nerve were traced with horseradish peroxidase. The commissural nucleus of the tractus solitarius was examined by dual labelling light and electron microscopy. Dense bilateral labelling with horseradish peroxidase was found in the tractus solitarius and commissural nucleus of the tractus solitarius. Horseradish peroxidase was found in unmyelinated axons, myelinated axons, and nerve terminals. About 88% of horseradish peroxidaselabelled carotid sinus nerve axons were unmyelinated. Tyrosine hydroxylase immunoreactivity was identified in unmyelinated axons, myelinated axons, dendrites, perikarya, and nerve terminals. Most tyrosine hydroxylase immunoreactive axons (93%) in the commissural nucleus of the tractus solitarius were unmyelinated. Tyrosine hydroxylase immunoreactivity was simultaneously identified in carotid sinus nerve unmyelinated axons, myelinated axons, and nerve terminals. These double-labelled terminals comprised 28% of the number of tyrosine hydroxylase immunoreactive terminals in the commissural nucleus of the tractus solitarius, and 55% of transganglionically-labelled terminals. Therefore, there are both central and peripheral sources of tyrosine hydroxylase immunoreactive nerve terminals in the commissural nucleus of the tractus solitarius. These data support the hypothesis that peripheral tyrosine hydroxylase immunoreactive neurons are involved in the origination of the chemoreceptor reflex. Axo-axonic synapses between peripheral carotid sinus nerve afferent terminals and central terminals containing tyrosine hydroxylase immunoreactivity were observed in 22% of the axo-axonic synapses observed. Thus, central tyrosine hydroxylase immunoreactivity neurons are involved in the modulation of the chemo- and/or baroreceptor reflexes. Synaptic contacts were not observed between carotid sinus nerve afferents and tyrosine hydroxylase immunoreactive perikarya of dendrites. Catecholaminergic neurons are thus unlikely to be the second order neurons of either the chemo- or baroreceptor reflex in the commissural nucleus of the tractus solitarius.     

Stretch receptor-associated expression of alpha 3 isoform of the Na+, K+-ATPase in rat peripheral nervous system

Expression of the neuronal alpha(3) isoform of the Na(+),K(+)-ATPase (alpha(3) Na(+),K(+)-ATPase) was studied in the rat peripheral nervous system using histological and immunohistochemical techniques. Non-uniform expression of the alpha(3) Na(+),K(+)-ATPase was observed in L5 ventral and dorsal roots, dorsal root ganglion, sciatic nerve and its branches into skeletal muscle. The alpha(3) Na(+),K(+)-ATPase was not detected in nerve fibers in skin, saphenous and sural nerves. In dorsal root ganglion 12+/-2% of neurons were immunopositive for alpha(3) Na(+),K(+)-ATPase and all these neurons were large primary afferents that were not labeled by Griffonia simplicifolia isolectin B4 (marker of small primary sensory neurons). In dorsal and ventral roots 27+/-3% and 40+/-3%, respectively, of myelinated axons displayed immunoreactivity for alpha(3) Na(+),K(+)-ATPase. In contrast to the dorsal roots, strong immunoreactivity in ventral roots was observed only in myelinated axons of small caliber, presumably gamma-efferents. In the mixed sciatic nerve alpha(3) Na(+),K(+)-ATPase was detected in 26+/-5% of myelinated axons (both small and large caliber). In extensor hallicus proprius and lumbricales hind limb muscles alpha(3) Na(+),K(+)-ATPase was detected in some intramuscular axons and axonal terminals on intrafusal muscle fibers in the spindle equatorial and polar regions (regions of afferent and efferent innervation of the muscle stretch receptor, respectively). No alpha(3) Na(+),K(+)-ATPase was found in association with innervation of extrafusal muscle fibers or in tendon-muscle fusion regions. These data demonstrate non-uniform expression of the alpha(3) isoform of the Na(+),K(+)-ATPase in rat peripheral nervous system and suggest that alpha(3) Na(+),K(+)-ATPase is specifically expressed in afferent and efferent axons innervating skeletal muscle stretch receptors.     

Segmental reflexes mediated by joint afferent neurons in cat knee

1. Reflex effects mediated by knee joint afferent neurons were tested using the electrically evoked monosynaptic reflex in motoneurons innervating muscles which act about the knee. 2. Joint afferents were excited using small extension movements of the knee, stimuli which have been shown to be effective in activating a large fraction of the myelinated fibers in joint nerves. Because of the variability of reflex responses observed by others under similar experimental conditions, experimental control and data collection were done automatically under computer control. Large numbers of trials were presented in random sequence, and the resulting data were sorted and averaged. 3. Positive feedback was observed in both knee extensor (vasti) and knee flexor (posterior biceps-semitendinosus (PB-ST)) motoneuron populations. Vasti motoneurons were excited, and PB-ST motoneurons were inhibited by knee extensions when the posterior articular nerve (PAN) was intact; these effects were lost or substantially reduced when the PAN was cut. 4. Using knee extension stimuli, no reliable reflex effects were observed which could be attributed to afferents in medial articular nerve.