Histone Variants as Stem Cell Biomarkers for Long-Term Injection Medialization Laryngoplasty.

OBJECTIVES/HYPOTHESIS: Vocal fold (VF) paralysis by sectioning the recurrent laryngeal nerve dramatically impacts the life of thyroidectomy patients. Volume-expanding materials can temporarily restore VF medialization. To prolong this benefit, adipose mesenchymal stem cells (ADSCs) and micronized acellular dermis (MACD) were co-injected in a rabbit model of injection medialization laryngoplasty. Biomarkers of in situ proliferation were identified by mass spectrometry proteomics and pathway analysis to guide future efforts to increase the length of benefit. METHODS: ADSCs were expanded and/or differentiated into chondrocytes (CHON) as collagen microspheres. After VF paralysis rabbits received MACD, MACD + undifferentiated ADSC, or MACD+CHON, ADSCs differentiated into chondrocytes. After 12 weeks, animals were sacrificed and 5-µm paraffin-embedded cryosections were prepared from larynges for hematoxylin and eosin visualization and nanoflow liquid chromatography electrospray-ionization tandem-mass spectrometry analysis of tissue collections. Validated proteins were processed by Venn subtraction and gene ontology (GO) overrepresentation analysis to identify unique pathways and biomarkers. RESULTS: Confirmed proteins numbered 147 (MACD), 1,243 (MACD+ADSC), and 1,033 (MACD+CHON). Totally, 333 proteins were uniquely found in the MACD+ADSC group, including mesenchymal surface markers CD9, CD44, fibronectin, and vimentin. Over 70% of proteins belonged to catalytic activity and binding GO categories, with the histone (H) family being overrepresented (P < 0.05). Histone variants H3.3, H2A.V, and H2A.Z (associated with open chromatin states) were overrepresented in the MACD+ADSC group, whereas structural histones H2A, H2B, and H4 were not. CONCLUSION: Biomarkers, including atypical histones, are associated with in vivo proliferation of ADSCs and an expanded VF medialization volume. LEVEL OF EVIDENCE: NA Laryngoscope, 128:E402-E408, 2018.

Morphologic evaluation of the fetal recurrent laryngeal nerve and motor units in the thyroarytenoid muscle.

This study is a morphologic description of the recurrent laryngeal nerve (RLN) and of the number and size of motor units (MUs) in the thyroarytenoid (TA) muscle bilaterally of a human fetus aged 25 weeks. A quantitative analysis of RLN and MUs is presented to investigate similarities with equivalent structures in adults. In the fetus used in our study, the morphologic organization of the RLN was similar to that commonly described in the adult RLN. Moreover, as is observed in adult TA, the TA of the analyzed fetus, particularly the right TA, showed MUs typical of muscles with great motor accuracy. These results may be used to increase our knowledge of the features of the voice in adults and newborns.

Recurrent laryngeal nerve transection and anastomosis: rat laryngeal motoneuron survival and effect of the anastomosis site.

OBJECTIVES: We investigated the quantity of recurrent laryngeal nerve motoneurons (RLNMs) that survive after transection and anastomosis of the rat recurrent laryngeal nerve (RLN), as well as the impact of the anastomosis site on RLN regeneration. METHODS: Ten rats underwent right RLN transection and anastomosis. After 16 weeks, Fluoro-Ruby (FR) was applied to the RLN that was transected proximal or distal to the anastomosis site. The brain stems were harvested, and the nucleus ambiguus was evaluated for labeled RLNMs. The RLNM counts were compared to each other and to those from 3 control rats in which FR was applied to an acutely transected RLN. RESULTS: The number of RLNMs that were stained after RLN transection, anastomosis, and regeneration was consistent with the total number of RLNMs in the nucleus ambiguus of control rats. This finding confirms that most RLNMs survived after RLN transection and anastomosis. The quantity of labeled RLNMs was statistically similar whether the FR was applied proximal or distal to the anastomosis, implying that most of the viable axons that were present proximal to the anastomosis crossed into the distal nerve. CONCLUSIONS: Rat RLNMs survive nerve transection, anastomosis, and regeneration. The anastomosis site does not significantly impede axonal regeneration, and most of the axons traverse the anastomosis into the distal nerve.

Reorganization of laryngeal motoneurons after crush injury in the recurrent laryngeal nerve of the rat.

Motoneurons innervating laryngeal muscles are located in the nucleus ambiguus (Amb), but there is no general agreement on the somatotopic representation and even less is known on how an injury in the recurrent laryngeal nerve (RLN) affects this pattern. This study analyzes the normal somatotopy of those motoneurons and describes its changes over time after a crush injury to the RLN. In the control group (control group 1, n = 9 rats), the posterior cricoarytenoid (PCA) and thyroarytenoid (TA) muscles were injected with cholera toxin-B. In the experimental groups the left RLN of each animal was crushed with a fine tip forceps and, after several survival periods (1, 2, 4, 8, 12 weeks; minimum six rats per time), the PCA and TA muscles were injected as described above. After each surgery, the motility of the vocal folds was evaluated. Additional control experiments were performed; the second control experiment (control group 2, n = 6 rats) was performed labeling the TA and PCA immediately prior to the section of the superior laryngeal nerve (SLN), in order to eliminate the possibility of accidental labeling of the cricothyroid (CT) muscle by spread from the injection site. The third control group (control group 3, n = 5 rats) was included to determine if there is some sprouting from the SLN into the territories of the RLN after a crush of this last nerve. One week after the crush injury of the RLN, the PCA and TA muscles were injected immediately before the section of the SLN. The results show that a single population of neurons represents each muscle with the PCA in the most rostral position followed caudalwards by the TA. One week post-RLN injury, both the somatotopy and the number of labeled motoneurons changed, where the labeled neurons were distributed randomly; in addition, an area of topographical overlap of the two populations was observed and vocal fold mobility was lost. In the rest of the survival periods, the overlapping area is larger, but the movement of the vocal folds tends to recover. After 12 weeks of survival, the disorganization within the Amb is the largest, but the number of motoneurons is similar to control, and all animals recovered the movement of the left vocal fold. Our additional controls indicate that no tracer spread to the CT muscle occurred, and that many of the labeled motoneurons from the PCA after 1 week post-RLN injury correspond to motoneurons whose axons travel in the SLN. Therefore, it seems that after RLN injury there is a collateral sprouting and collateral innervation. Although the somatotopic organization of the Amb is lost after a crush injury of the RLN and does not recover in the times studied here, the movement of the vocal folds as well as the number of neurons that supply the TA and the PCA muscles recovered within 8 weeks, indicating that the central nervous system of the rat has a great capacity of plasticity.

Electromyographic and histologic evolution of the recurrent laryngeal nerve from transection and anastomosis to mature reinnervation.

OBJECTIVES: To describe the natural evolution of recurrent laryngeal nerve (RLN) reinnervation in an animal model. STUDY DESIGN: Twenty Sprague Dawley rats underwent unilateral RLN transection and anastomosis. Animals were sacrificed at 4, 8, 12, 16, and 20 weeks. Prior to sacrifice, each rat underwent electromyography (EMG) and visual grading of vocal fold motion. Bilateral RLNs were harvested and evaluated histologically. RESULTS: EMG revealed synkinetic reinnervation at all time periods except at 4 weeks. EMG evolution plateaued at 16 weeks. Vocal fold motion was slight in three rats at 4 weeks but was otherwise absent except for one rat at 12 weeks. Histologic changes of the axons and their myelin sheaths were consistent at each time period. At 16 weeks, histologic changes plateaued. CONCLUSIONS: Consistent EMG, histologic, and vocal fold motion changes occur at specific time periods during RLN reinnervation after transection and anastomosis in a rat model. Reinnervation is mature at 16 weeks. Findings corroborate theories of preferential and synkinetic reinnervation after RLN transection. Use of a rat model to investigate the effect of interventions on RLN reinnervation requires a minimum of 16 weeks between transection and investigation to allow for maturation of reinnervation.

Collateral reinnervation by the superior laryngeal nerve after recurrent laryngeal nerve injury.

This study investigates the role of the intact superior laryngeal nerve (SLN) in the reinnervation process of one of the laryngeal muscles, the posterior cricoarytenoid muscle (PCA), following recurrent laryngeal nerve (RLN) injury. Using a chronic RLN injury model in the adult rat, PCA reinnervation was assessed by retrograde double-tracing techniques in combination with electrophysiology and immunohistochemistry of muscle sections. The results demonstrate that the PCA receives dual innervation from both laryngeal nerves even in the uninjured system. Functionally significant collateral reinnervation originates from intact SLN fibers following RLN injury, mainly due to intramuscular sprouting rather than by recruitment of more motor neurons. This may be important when choosing surgical and/or medical treatment for patients with RLN injury.

Morfometría de los nervios laríngeos recurrentes de la rata / Morphometry of the recurrent laryngeal nerves of the rat

Los nervios laríngeos recurrentes (NLR) de los mamíferos son diferentes en longitud entre ambos lados. Esta asimetría implica, a priori, diferentes tiempos de conducción del estímulo a la musculatura laríngea controlada por cada nervio, postulándose diversos modelos para explicar el cierre glótico sincrónico más allá de la citada diferencia. Varios son los estudios publicados en esta materia aunque, no obstante, presentan carencias en datos relevantes. Utilizando dos grupos de 10 y 6 ratas, respectivamente, nuestro estudio compara la longitud de los NLR por lado y, mediante microscopía óptica acoplada a un sistema de análisis morfométrico, el número y características de las fibras mielínicas que los componen. Los resultados muestran que el NLR izquierdo (NLRi) es, de promedio, 0,84 cm más largo que el NLR derecho (NLRd). No hay diferencias estadísticamente significativas en el número de fibras por lado pero sí en el grosor de las mismas, mayores en el NLRd. Estos datos se analizan valorando los posibles mecanismos de compensación de la diferencia de longitud de los NLR

In mammals the recurrent laryngeal nerves are dissimilar in length between both sides. This asymmetry involves different time of arrival of the stimulus to the laryngeal musculature controlled by each nerve. Thus, several explanations have been addressed to elucidate the closest of the glottis at the same time despite the unlike length of the nerves. However, previous works on the topic lack of several important data. The present study compares, in two groups of 10 and 6 rats, the length and the composition of myelinated fibers in the recurrent laryngeal nerves of both sides, by means of light microscopy and a computerized morphometric analysis. The results show a mean difference of 0,84 cm longer the left than the right recurrent laryngeal nerve. No statistical differences were observed in the number of myelinated fibers between both sides. However, the myelinated fibers of the right side were statistically bigger in diameter than the fibers of the left side. The data are discussed in the context of the mechanisms for the compensation of the dissimilar length of both recurrent laryngeal nerves

[Morphometry of the recurrent laryngeal nerves of the rat]. / Morfometría de los nervios laríngeos recurrentes de la rata.

In mammals the recurrent laryngeal nerves are dissimilar in length between both sides. This asymmetry involves different time of arrival of the stimulus to the laryngeal musculature controlled by each nerve. Thus, several explanations have been addressed to elucidate the closest of the glottis at the same time despite the unlike length of the nerves. However, previous works on the topic lack of several important data. The present study compares, in two groups of 10 and 6 rats, the length and the composition of myelinated fibers in the recurrent laryngeal nerves of both sides, by means of light microscopy and a computerized morphometric analysis. The results show a mean difference of 0,84 cm longer the left than the right recurrent laryngeal nerve. No statistical differences were observed in the number of myelinated fibers between both sides. However, the myelinated fibers of the right side were statistically bigger in diameter than the fibers of the left side. The data are discussed in the context of the mechanisms for the compensation of the dissimilar length of both recurrent laryngeal nerves.

Laryngeal abductor muscle reinnervation in a pig model.

OBJECTIVE: To develop a large animal model for studies of laryngeal abductor reinnervation. MATERIAL AND METHODS: Six minipigs underwent unilateral anastomosis of the phrenic nerve-abductor branch of the recurrent laryngeal nerve (RLN). Polyhydroxybutyrate (PHB) conduits were used for repair. At each of 30, 60 and 120 days, 2 animals underwent video laryngeal endoscopy (VLE) and were then killed. VLE was also performed in the 120-day pair at 60 days. Nerve-conduit-nerve-muscle samples were fixed for light and immunofluorescence (pan-neurofilaments, S-100) microscopy. Laryngeal muscles were harvested (myosin heavy chain analysis). RESULTS: VLE showed recovery of abductor function in 1 animal at 60 days and in 1 at 120 days. Haematoxylin-eosin staining demonstrated a complex inflammatory response. Eosinophil recruitment was observed. Stepwise regeneration and reorganization of the distal nerve between 30 and 120 days was observed with pan-NF staining. The mean minimum diameter in the reinnervated posterior crico-arytenoids tended to increase for up to 120 days. CONCLUSIONS: Anastomosis of the phrenic nerve-abductor branch of the RLN with a PHB conduit in a pig can result in functional and histological recovery within 2-4 months and appears to at least sustain abductor muscle fibre morphology. Recovery occurs despite a complex inflammatory response, which may be an essential part of healing rather than inhibitory.

Recurrent laryngeal nerve regeneration by tissue engineering.

The recurrent laryngeal nerve (RLN) does not regenerate well after it has been cut, and no current surgical methods achieve functional regeneration. Here, we evaluate the functional regeneration of the RLN after reconstruction using a biodegradable nerve conduit or an autologous nerve graft. The nerve conduit was made of a polyglycolic acid (PGA) tube coated with collagen. A 10-mm gap in the resected nerve was bridged by a PGA tube in 6 adult beagle dogs (group 1) and by an autologous nerve graft in 3 dogs (group 2). Fiberscopic observation revealed functional regeneration of the RLN in 4 of the 6 dogs in group 1. No regeneration of the RLN was observed in any dog in group 2. We also tested for axonal transport, and measured the compound muscle action potential. The RLN can be functionally regenerated with a PGA tube, which may act as a scaffold for the growth of regenerating axons.

Glycinergic inhibition is essential for co-ordinating cranial and spinal respiratory motor outputs in the neonatal rat.

Eupnoeic breathing in mammals is dependent on the co-ordinated activity of cranial and spinal motor outputs to both ventilate the lungs and adjust respiratory airflow, which they do by regulating upper-airway resistance. We investigated the role of central glycinergic inhibition in the co-ordination of cranial and spinal respiratory motor outflows. We developed an arterially perfused neonatal rat preparation (postnatal age 0-4 days) to assess the effects of blocking glycine receptors with systemically administered strychnine (0.5-1 microM). We recorded respiratory neurones located within the ventrolateral medulla, inspiratory phrenic nerve activity (PNA) and recurrent laryngeal nerve activity (RLNA), as well as dynamic changes in laryngeal resistance. Central recordings of postinspiratory neurones revealed an earlier onset in firing relative to the onset of inspiratory PNA after exposure to strychnine (260 +/- 38.9 vs. 129 +/- 26.8 ms). After glycine receptor blockade, postinspiratory neurones discharged during the inspiratory phase. Strychnine also evoked a decrease in PNA frequency (from 38.6 +/- 4.7 to 30.7 +/- 2.8 bursts min(-1)), but amplitude was unaffected. In control conditions, RLNA comprised inspiratory and postinspiratory discharges; the amplitude of the latter exceeded that of the former. However, after administration of strychnine, the amplitude of inspiratory-related discharge increased (+65.2 +/- 15.2 %) and exceeded postinspiratory activity. Functionally this change in RLNA caused a paradoxical, inspiratory-related glottal constriction during PNA. We conclude that during the first days of life in the rat, glycine receptors are essential for the formation of the eupnoeic-like breathing pattern as defined by the co-ordinated activity of cranial and spinal motor inspiratory and postinspiratory activities.

Enhanced expression of neuronal nitric oxide synthase and phospholipase C-gamma1 in regenerating murine neuronal cells by pulsed electromagnetic field.

Pulsed electromagnetic field (PEMF) has been shown to improve the rate of peripheral nerve regeneration. In the present study we investigated the expression of neuronal nitric oxide synthase (nNOS) and phospholipase C-gamma1 (PLC-gamma1) in regenerating rat laryngeal nerves during the exposure to PEMF after surgical transection and reanastomosis. Axons were found to regenerate into the distal stump nearly twice faster in PEMF-exposed animals than in the control. Consistently, motor function was better recovered in PEMF-treated rats. The expression of nNOS and PLC-gamma1 was highly enhanced in the regenerated nerves.

Pulmonary C-fiber activation enhances respiratory-related activities of the recurrent laryngeal nerve in rats.

The purpose of the current study was to characterize the response of the recurrent laryngeal nerve (RLN) to pulmonary C-fiber activation. Male rats of Wistar strain were anesthetized by urethane (1.2 g/kg, i.p.). Tracheostomy was performed. Catheter was inserted into the femoral artery and vein. Additional catheter was placed near the entrance of the right atrium via the right jugular vein. The animal was then paralyzed with gallamine triethiodide, ventilated and maintained at normocapnia in hyperoxia. Activities of the phrenic (PNA) and recurrent laryngeal nerves (RLNA) were monitored simultaneously. Two experimental protocols were completed. In the first experiment, various doses of capsaicin were delivered into the right atrium to activate pulmonary C-fibers with vagal intact. Low dose of capsaicin (1.25 microg/kg) produced apnea, a decrease in amplitude of PNA, an enhancement of RLNA during apnea and recovery from apnea, hypotension, and bradycardia. High dose of capsaicin (5 and 20 microg/kg) evoked the same tendency of response for both nerves and biphasic changes in blood pressure. Dose dependency was only seen in the period of apnea but not observable in nerve amplitudes. After bilateral vagotomy, low dose of capsaicin produced an increase in PNA without apnea, no significant change in RLNA, and hypertension. These results suggest that activation of vagal and nonvagal C-fibers could produce different reflex effects on cardiopulmonary functions. The reflex responses evoked by these two types of afferents might play defensive and protective roles in the airways and lungs.

Morphology and electrophysiology of guinea-pig paratracheal neurones.

BACKGROUND: Although guinea-pig tracheal preparations are used as models of asthma, the morphological and electrophysiological characteristics of its associated ganglion neurones (paratracheal neurones) have not been characterized. METHODS: Intracellular staining and electrophysiological recording techniques have been applied to guinea-pig paratracheal neurones in isolated preparations. RESULTS: Most (32/35) neurones were multipolar, with many short (< 70 microns), finely tapering processes and one or more long processes; the latter, which were traced for up to 400 microns, travelled along the interconnecting nerve trunks, often in pairs, or over smooth muscle bundles. About 20% (6/32) of neurones had conspicuous somal extensions that gave rise to 3-8 processes. The soma morphology of neurones of the intrinsic ganglionated plexus close to the trachealis muscle were usually more complex than those in or associated with recurrent or vagal nerve trunks. Two types of neurone were identified electrophysiologically; neurones with fast excitatory synaptic potentials were found only in ganglia located very close to the smooth muscle, whereas > 90% of neurones lacking synaptic inputs were associated with recurrent nerve trunks. Transmural or focal electrical stimulation failed to evoke either slow inhibitory or slow excitatory (cholinergic or non-cholinergic) synaptic potentials in either electrophysiological type. CONCLUSIONS: It is tentatively concluded that the neurones of the intrinsic ganglionated plexus receiving synaptic input probably provided the para-sympathetic innervation to effector cells (such as trachealis muscle). Both these and the spiking neurones located in or near nerve trunks showed little potential for synaptic modulation of their excitability.

Properties of postganglionic sympathetic fibers isolated from the right recurrent laryngeal nerve.

The pattern of response of 45 single postganglionic sympathetic axons dissected from the right recurrent laryngeal nerve was examined in chloralose-anesthetized cats. Both vagoaortic nerves were cut, and both sinus nerves were left intact. Each neuron, based on the presence of cardiac and respiratory rhythmicities in its resting activity and reaction to systemic hypoxia (10% O2 in N2 for 2 min), was classified into one of three classes. Class I neurons (n = 29, 64%) were activated during systemic hypoxia and had a pronounced cardiac and inspiration-related rhythmicity in their resting activity. Class II neurons (n = 12,27%) were inhibited during systemic hypoxia, and their cardiac and respiratory rhythmicities were either negligible or totally absent. Class III neurons (n = 4,9%), similarly to class I, had a pronounced cardiac and inspiratory rhythmicity but were not affected by systemic hypoxia. The systemic hypoxia was always accompanied by an increase in blood pressure. We conclude that class I and possibly class III neurons innervate the arteries of upper airways. We also discuss the possibility that class II neurons are responsible for regulating the smooth muscles of upper airways.

Morphometric study of the recurrent laryngeal nerve in young 'normal' horses.

Quantitative measurements were made on cross-sectional preparations of the distal part of the recurrent laryngeal nerve (RLN) from nine young mixed-breed horses to establish reference values for the total number of myelinated fibres, mean fibre diameter and percentage of thickest fibres (over 9.5 microns) and to delineate diameter distribution curves. The total number of myelinated fibres, mean fibre diameter and percentage of thickest fibres for the left RLN were significantly lower than those of the right RLN (P < 0.005). The distribution of fibres was unimodal. The fibre diameter ranged from 1 micron to 17 microns. Approximately, 95 per cent of fibres had a diameter larger than 5 microns.

Phrenic and recurrent laryngeal motoneuron activities during hyperoxia and hypoxia in piglets.

We hypothesized that synchronization of inspiratory motoneurons may involve inputs from two central pattern generators (CPG): one characterized by medium-frequency (< 50 Hz) and the other by high-frequency oscillations (> or = 50 Hz). We studied phrenic and recurrent laryngeal nerve activities recorded during hyperoxia and hypoxia in Saffan anesthetized, paralyzed, and artificially ventilated piglets. Spectral analyses, derived from the full as well as partitioned halves of inspiration, showed that phrenic and recurrent laryngeal discharges contained peaks in the medium-frequency band, which were indicative of common inputs. The phrenic spectra of many animals had peaks in the high-frequency band; such peaks were uncommon in recurrent laryngeal spectra; consequently, correlated activities corresponding to high-frequency oscillations were not usually observed. Thus, it is likely that acquisition of modulating inputs from a high-frequency CPG may emerge in an age-dependent manner in different motoneuron pools. During hypoxia, both phrenic and recurrent laryngeal discharges were facilitated as shown by increases in both the amplitudes of signal-averaged histograms and the magnitudes of their respective power spectral activities. Also, there was a significant increase in the values of phrenic-recurrent laryngeal coherence estimates in the medium-frequency region. Hence, medium-frequency oscillations are more apparent in early development, perhaps to facilitate synchronization of inspiratory motoneuron activities, especially under conditions of increased chemical drive.

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.

Morphologic and morphometric studies of the vagus and recurrent laryngeal nerves in clinically normal adult dogs.

Morphologic and morphometric studies were performed on the vagus nerve and its major branch, the recurrent laryngeal nerve (RLN): (1) to determine normal histologic data in myelinated fibers of clinically normal young adult dogs, (2) to establish reference values for mean fiber diameter in the vagus nerve and in the proximal and distal portions of the RLN, and (3) to delineate relative frequency distribution curves for each nerve. Few degenerative changes were observed in single teased-nerve fiber preparations. There was no statistical difference between left and right sides of the vagus nerve or between the proximal and distal portions of the RLN (right and left sides). In contrast to the unimodal distribution of fibers in the vagus nerve and the proximal portion of the RLN, the distribution of fibers in the distal portion of the RLN was bimodal. Mean (+/- SD) fiber diameters of the vagus nerve and in the proximal portion of the RLN (3.02 +/- 1.44 microns and 3.63 +/- 1.49 microns, respectively) were not significantly different, despite a shift to large-diameter fibers in the latter. However, mean fiber diameters of distal and proximal portions of the RLN (5.56 +/- 1.88 microns and 3.63 +/- 1.49 microns, respectively) were significantly (P less than 0.001) different. Approximately 86% of fibers in the vagus nerve and 76% of fibers in the proximal portion of the RLN, had a diameter less than 5 microns, and about 70% of fibers in the distal portion of the RLN had a diameter greater than 5 microns.(ABSTRACT TRUNCATED AT 250 WORDS)

Lack of size-related differential sensitivity to equilibrium conduction block among mammalian myelinated axons exposed to lidocaine.

This study sought to evaluate the sensitivity of individual, relatively thick myelinated axons of mammalian nerve to equilibrium conduction block by lidocaine, and to compare this to the incidence of conduction block previously measured in individual thinner myelinated axons. The incidence of conduction block by lidocaine 0.3 and 0.6 mM (8.1-16.2 mg/dl) was determined on 35 individual axons in dissected filaments of rabbit recurrent laryngeal nerve (RLN) in which the control conduction velocity ranged from 28 to 77 m/sec. Thirty-four axons (97%) remained excitable in lidocaine 0.3 mM; 2 axons (6%) remained excitable in lidocaine 0.6 mM. These proportions did not differ significantly (P greater than 0.2) from those in thinner axons of vagus, either in comparisons with previous data from extracellular recordings or with new data from filament recordings. The results imply that differential blocks observed under clinical conditions probably depend on factors other than a size-related difference in the minimal equilibrium blocking concentration among myelinated axons. Equilibrium depression of the amplitude of RLN compound action potentials by lidocaine 0.3 and 0.6 mM was disproportionately great relative to the incidence of equilibrium conduction block in individual axons, confirming that depression of the compound action potential is not a reliable measure of nerve conduction block.