Morphometric Comparison between Controlateral Sciatic Nerves in the Male and Female Rabbit

Some morphometrical parameters of the axons making up the controlateral sciatic nerves, both in the male and female rabbits were calculated and compared by means of a Zeiss Vidas image analyser (Ober Kochen, Germany). The results show that the fibres constituting the left nerve have a greater mean diameter but a lower mean density than those constituting the right nerve. This suggests that the diameter of the myelinated fibres and the density of both the myelinated and unmyelinated fibres do not vary from male to female. On the other hand, the G ratio and the diameter of the unmyelinated axons do, since the nerves on the right side (in both sexes) have higher morphometric values, on average, than the controlateral ones.     

Morphometric Changes in the Dog Trochlear Nerve with Growth

The objective of this work was to analyse changes in morphometric characteristics related to growth in the trochlear nerve in dogs. Twenty beagles, split into four dog age groups (A, 7 days; B, 21 days; C, 35 days; D, 49 days and E, 4 years), were used. The right intracranial portion of the nerve was analysed by light and electron microscopy. The nerve cross‐sectional area was calculated. Number, diameter and cross‐sectional area of unmyelinated and myelinated fibres were also calculated. In myelinated fibres, the corresponding axon area and diameter and myelin sheath thickness were also calculated. The number of myelinated and unmyelinated fibres was 1070.25 ± 112.07 and 592.25 ± 467.53 in group A, 1367 ± 57.98 and 143.67 ± 54.37 in group B, 1574.20 ± 299.50 and 151.67 ± 51.73 in group C, 1340.33 ± 151 and 127 ± 48.75 in group D and 1476 ± 260.71 and 284 ± 101.82 in group E. The mean diameter for myelinated and unmyelinated fibres was 4.37 ± 0.17 μm and 0.41 ± 0.08 μm for group A; 6.21 ± 0.12 μm and 0.30 ± 0.03 μm for B; 6.90 ± 0.91 μm and 0.32 ± 0.03 μm for C; 7.86 ± 1.19 μm and 0.32 ± 0.02 μm for D; 10.63 ± 0.50 μm and 0.30 ± 0.01 μm for E, respectively. This nerve possesses similar structural and ultrastructural features to the same nerve in other species and modifies its morphometry with growth. Results could enhance the understanding of pathological disorders.     

Quantitative analysis of the optic nerve of the horse (Thoroughbred)

Three optic nerves (L1, R2, R3) 12-18 mm behind the eyeball of the horse (Thoroughbred) were investigated quantitatively under light and electron microscopes. Thin sections at the thickness of 0.35 microm were cut, stained by toluidine blue and observed under the light microscope. The areas of the optic nerve and the axon bundles were 20.03 +/- 1.04 and 16.59 +/- 0.79 mm2 (mean +/- SD, n=3), respectively. The axon numbers for optic nerve L1, R2 and R3, estimated from light micrographs, were about 481 x 10(3), 543 x 10(3), and 494 x 10(3), respectively. Axons of optic nerve L1 were also counted from electron micrographs and the total number of 488 x 10(3) was received. Furthemore, axon diameters of optic nerve L1 were also measured from electron micrographs. The diameter of a circle with the same peripheral length as an axon, was regarded as its diameter. The medullary sheath of the axon was not included during measuring. Altogether 5,744 axons were measured and axon diameters were in a range of 0.23-12.69 microm, with a mean of 2.56 +/- 1.45 microm (mean +/- SD). A regional difference of axonal diameters was found across the optic nerve: the mean diameter of axons in the centrodorsal region (2.28 microm) was the smallest, and had significant difference with those in several peripheral regions (P<0.05).     

A sensory neuropathy affecting Long-haired Dachshund dogs

A sensory neuropathy affecting three Long-haired Dachshunds is described. The clinical signs were similar and include disturbances of proprioception, pain sensation and urinary function. Sensory nerve conduction was absent in one dog. Nerve biopsy showed a severe depletion of myelinated fibres in sensory nerves and widespread unmyelinated fibre pathology. Clinical and pathological evidence suggests that this axonal neuropathy, which is possibly of inherited origin, affects both myelinated and unmyelinated fibres in sensory and autonomic nerve trunks.     

Quantitative studies of the optic nerve fiber layer in the chicken retina.

The optic nerve fiber layer (NFL) of the chicken retina was studied quantitatively and morphologically at 17 positions along seven radially arranged bands from the dorsal tip of the pecten oculi using electron microscopy. The number of nerve fibers was counted in areas 6 microm in width x the full thickness of the NFL. Myelinated nerve fibers in the NFL were also identified immunohistochemically using anti-myelin basic protein serum. The dorsal area (dorsal, dorso-temporal and dorso-nasal bands) in the retina had thin NFL and contained the largest number of nerve fibers, which were mainly thin and unmyelinated. The ventral area (ventral and ventro-temporal bands) had a thick NFL and contained a relatively small number of nerve fibers, many of which were myelinated. The nasal band had the thickest NFL and contained as many nerve fibers as the dorsal area, with the temporal band showing a high ratio of myelinated fibers. The band had a thick NFL and contained many nerve fibers with a relatively low ratio of myelinated fibers. The relationship between the number and composition of nerve fibers in the NFL to the chicken visual characteristics was discussed. Although the myelin in the chicken retina was loose type, the myelin-forming cells were similar in appearance to dense oligodendrocytes. retina, morphometry, myelinated fiber, nerve fiber layer.     

Pathological aspects of Australian Stringhalt.

Nine horses with clinical signs of Australian Stringhalt were killed and tissues collected for a detailed pathological study. Lesions were limited to peripheral nerves and muscles. The most severely affected nerves were the superficial and deep peroneal, distal tibial, plantar digital, volar and recurrent laryngeal nerve with changes characterised by a selective loss of large diameter myelinated fibres with various degrees of demyelination, fibrosis, Schwann cell proliferation and onion-bulb formation. A routine evaluation of the brain and spinal cord by light microscopy failed to reveal any consistent abnormalities. Morphometric analysis of deep peroneal and recurrent laryngeal nerves confirmed the reduced number of large diameter myelinated axons. Teased fibre preparations of these nerves did not show any abnormalities in internodal distance. The most severe muscle lesions were in the long and lateral digital extensors, cranial tibial, dorsal cricoarytenoid, gracilis and lateral deep digital flexor with extensive atrophy of fibres and diffuse fibrosis. Histochemical evaluation of the long digital extensor from 3 affected horses showed an abnormally wide distribution in fibre size and a reduction in type II fibres compared with controls. These lesions are consistent with a distal axonopathy leading to neurogenic muscle atrophy. The distribution of neuromuscular lesions in Australian Stringhalt may be explained by the susceptibility of longer, larger myelinated nerve fibres to injury, but the cause for this distal axonopathy remains unknown.     

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 gm) 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 μm to 17 μm. Approximately, 95 per cent of fibres had a diameter larger than 5 μm.     

Morphologic and morphometric study of the facial nerve in clinically normal adult dogs.

A morphologic and morphometric study was carried out on the facial nerve: to determine normal histologic data in myelinated fibers of clinically normal young adult dogs; to establish reference values for mean fiber diameter, and to delineate the relative diameter frequency distribution curve. Few degenerative changes were observed in single teased-nerve fibers and semithin cross-sectional nerve preparations. Statistical difference was not observed between left and right facial nerves. The distribution of fiber diameters in the facial nerve was unimodal. Mean (+/- SD) fiber diameter of the facial nerve was 3.92 +/- 1.18 microns. Approximately 89% of fibers in the facial nerve had diameter between 3 and 6 microns. Fiber diameter ranged from 2 to 12 microns; however, less than 1% of fibers had diameter greater than 8 microns.     

Fine structure of the retino-optic nerve junction in dogs

In most mammals, the optic nerve fibers are myelinated in its extraocular part (EON) but not in its intraocular part (ION) and also in the retina. Transitional zone from the myelinated to unmyelinated optic nerve usually lies in the central part to the lamina cribrosa. It has been known that dogs contain exceptionally myelinated fibers in ION by light microscopy. The aim of this study was to investigate electron microscopically the retino-optic nerve junction in dogs and re-evaluate the barrier to migration of oligodendroblasts into ION. Fourteen adult dogs were used. EON was largely myelinated. In ION the percentage of myelinated fibers decreased gradually toward the retina. A narrow area of ION adjoining the retina was completely unmyelinated. In most mammalian optic nerves, oligodendrocytes are not found in ION. It has been suggested that oligodendroblasts are prevented from migrating from EON into ION; that is to say, there is a barrier to migration of oligodendroblasts. The lamina cribrosa, a dense meshwork of fibrous astrocytic processes, and a defect in the blood optic nerve barrier have been proposed as a candidate for the barrier to migration. Our results suggest, however, that these factors, at least in dogs, would be not involved in the formation of a barrier to migration of oligodendroblasts.     

Equine laryngeal hemiplegia. Part II. An electron microscopic study of peripheral nerves

The recurrent laryngeal nerves were examined by electron microscopy in five control, four subclinical and four clinical laryngeal hemiplegic horses. In addition, the peroneal nerve was examined in two horses in the latter group. The distally distributed loss of large myelinated fibres in the left recurrent laryngeal nerve seen by light microscopy was confirmed. In addition, active axonal pathology was found to be more evident than indicated by light microscopic investigations. The onion bulb formations observed indicated the repetitive nature of the damaging influence to nerve fibres. Although the pathological changes were most obvious in the distal left recurrent laryngeal nerve, alterations similar in type and distribution were present in other areas of the left and right nerves, and in the distal hindlimb nerves. The observation of fibres with inappropriately thick myelin sheaths relative to their axonal calibre, was confirmed statistically by determining the regressions of axis cylinder perimeter against the number of myelin lamellae. In conclusion, the peripheral nerve pathology of equine laryngeal hemiplegia was demonstrated to be a distally distributed loss of myelinated fibres, with considerable active axonal damage, in conjunction with axonal atrophy. These features suggest that this disease may be classified as a distal axonopathy.     

Preferential denervation of the adductor muscles of the equine larynx. II: Nerve pathology

The terminal branches of the recurrent laryngeal nerve (RLN) of three normal ponies and six horses with sub-clinical laryngeal disease were examined qualitatively and quantitatively in an attempt to explain the preferential denervation of the laryngeal adductor muscles in the neuropathy of idiopathic laryngeal hemiplegia (ILH). The myelinated fibre spectra of all the motor nerve fibres in the left and right abductor and adductor branches of the RLN in three normal ponies were measured. The density of myelinated fibres was also calculated. There was no significant difference between the larger group of myelinated fibres in the adductor or abductor branches. In the six horses with laryngeal hemiparesis, however, there was a marked preferential loss of the medium/large size myelinated fibres in the left adductor branch, although nerve fibre densities were not significantly different. While no simple morphometrical feature was found to explain the selective muscle denervation, the greater loss of large diameter myelinated fibres in the adductor branches confirms the earlier observation of adductor muscle susceptibility in the neuropathy of ILH.     

Equine laryngeal hemiplegia. Part I. A light microscopic study of peripheral nerves

This light microscopic investigation of 15 Thoroughbred horses provided substantial evidence for the classification of equine laryngeal hemiplegia as a distal axonopathy. Morphologic and morphometric examinations were performed on resin embedded recurrent laryngeal nerves from control, subclinical and clinical laryngeal hemiplegic animals. In the latter group of animals some distal hindlimb nerves were also examined. A distally graded loss of myelinated fibres selectively affecting those of large diameter was demonstrated in both left and right recurrent laryngeal nerves. Morphologic evidence of similar pathological changes in long hindlimb nerves was also present. An explanation for the early involvement of the cricoarytenoideus lateralis muscle in the course of laryngeal hemiplegia, was offered by the demonstration of more large diameter fibres in the branch of the recurrent laryngeal nerve innervating it, thus making it more susceptible to the disease process.     

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)     

Stereological characteristics of the equine accessory nerve

Stereological techniques have been increasingly employed for assessment and characterization of neuromuscular diseases in humans and animals. As an adjunct to histopathology, morphometrical algorithms provide quantitative evidence of the peripheral nerve composition, thereby shedding light on its fibre characteristics and basic electrophysiological properties. In the horse, stereological investigations already have focussed on the recurrent laryngeal, deep peroneal and lateral palmar nerves (LPN). Of these, only the latter is suitable for taking biopsies in clinical settings, however, it does not contain any motor fibres and Ia-afferents. On account of its virtually mixed fibre qualities, most researchers today recommend the cervical branch of the equine accessory nerve (AN) for harvesting diagnostic samples. Thus, the present study was carried out to gain morphometrical proof of the AN composition and to obtain stereological base values in healthy individuals using state-of-the-art technology. All parameters were compared to the common peroneal nerve (CPN), known to harbour all myelinated fibre classes. As this second biopsy site is located farther distally to the neuro-axis, attention was paid to possible length-dependent features. Taken together, digital image analysis could be accurately applied on all AN samples. Stereology supported the histological and clinical evidence that the AN contains all myelinated fibre types. The huge range and scatter of fibre counts and density (3351-17,812/mm(2)) per fascicle were comparable to that measured in the equine common peroneal, deep peroneal, lateral palmar and recurrent laryngeal nerves. Similar to those, fibre diameter distribution was bimodal with slow Abeta- and Agamma-mechanoceptor afferents outnumbering large myelinated Aalpha-fibres by a factor of about 1.5. With a g-ratio at 0.55 +/- 0.001, the overall degree of myelination in the AN is highly consistent and insignificantly ranges between that of the equine common peroneal and LPNs. Apart from this subtle deviation, a statistically relevant difference between the more proximal AN and the distal CPN could not be documented. By obtaining morphometrical standard parameters and even more sophisticated distribution indices, stereology is a valuable tool for detection of subtle changes that are likely to escape from the investigators' eyes. The AN serves as a reliable source for advanced peripheral nerve research and should be accompanied by farther distal nerve probes for assessment of neuropathies that present with a proximodistal gradient.     

Growth Dynamics of White Muscle Fibres in Relation to Somatic Growth of Larvae of European Sea Bass (Dicentrarchus labrax, L.)

The body length and diameter of muscle fibres from the right hypaxial tail myomeres were measured in 100 sea bass larvae (Dicentrarchus labrax, L.) at ages of 40 to 90 days, from a fish farm in Croatia. The larvae were kept at a temperature of 19-20 degrees C and fed with standard feed for their respective ages. Body lengths of larvae 40-90 days old varied from 8 to 42 mm, increasing significantly in 10-day intervals (p < 0.001), except between days 50 and 60 when there was no increase in the body length. The measured diameters of muscle fibres ranged from 10 to 80 microm, the mean value increasing except between days 50 and 60. The fibre diameter in larvae aged up to 60 days was 10-40 microm, while that in larvae older than 70 days was 51-80 microm. In 90-day-old larvae there were no fibres of 10-20 microm in diameter; however, the number of fibers of 61-80 microm diameter increased. A statistically significant concordance was determined between the body length and fibre diameter of larvae 70 to 90 days old. Between days 50 and 60, arrested growth of the larval body and corresponding decrease in fibre diameter was observed. From day 45 until days 59, larvae were fed with Artemia nauplii and microparticles of formulated diets. From the day 60, minced fish was added to formulated diets. This change in diet probably resulted in the observed growth arrest.     

Assessment of dorsal nerve root and spinal cord dorsal horn function in clinically normal dogs by determination of cord dorsum potentials

OBJECTIVE: To establish normal predictive values for cord dorsum potential (CDP) onset latency after thoracic and pelvic limb sensory or mixed nerve stimulation in adult dogs. ANIMALS: 26 clinically normal adult dogs. PROCEDURE: Sensory nerve action potentials (SNAP) were recorded proximally from tibial and lateral superficial radial nerves after distal stimulation. The CDP were recorded from the L4-L5 interarcuate ligament for the tibial nerve and from the C7-T1 interarcuate ligament for the radial nerve. Linear regression analyses were performed for CDP onset latency, and mean +/- SD was calculated for CDP onset to peak latency differences and sensory nerve conduction velocities (SNCV). RESULTS: For the tibial nerve, expected CDP onset latency (CDPOL) = -1.194 + 0.014 X pelvic limb length (mm; R2 = 0.912); CDPOL = -2.156 + 0.011 X pelvic limb/spinal length (mm; R2 = 0.911); and CDPOL = 0.941 + 2.197 X tibial nerve SNAP latency (milliseconds; R2 = 0.903). For the radial nerve, CDPOL = -0.9 + 0.014 x thoracic limb length (mm; R2 = 0.873); and CDPOL = 1.454 + 1.874 X radial nerve SNAP latency (milliseconds; R2 = 0.903). Mean +/- SD for CDP onset to peak latency difference for tibial and radial nerves was 3.1+/-0.3 and 3.0+/-0.4 milliseconds, respectively. CONCLUSIONS: Strong linear associations exist between CDPOL and a number of easily measured peripheral independent variables in dogs. There is also a narrow range of normal values for CDP onset to peak latency differences that is independent of limb length. CLINICAL RELEVANCE: CDP evaluation can be used to accurately assess functional severity and distribution of abnormalities in proximal sensory nerves, dorsal nerve roots, and spinal cord dorsal horns in dogs with suspected neuropathy, radiculopathy, or myelopathy involving the brachial or lumbosacral intumescences.     

Distribution of peptidergic nerve fibres in the guinea pig trabecular meshwork

A quantitative analysis of peptidergic nerve fibres located in the trabecular meshwork of the guinea pig has been performed. Our results confirm that this structure contains VIP-, NPY- and substance P-like immunoreactivity as major neurotransmitters. These findings were obtained using immunohistochemical techniques. For this purpose serial sections of the eye were stained by immunohistochemistry for each of three neurotransmitters and stained sections were analysed by quantitative image analysis. Our findings demonstrate that SP-positive, NPY-positive and VIP-positive nerve fibres occupy 11.2, 4.9 and 2.4%, respectively, of the observed area (expressed as conventional units, C.U.) in the trabecular meshwork of the guinea pig eye. It is relevant to emphasize that the area containing these three types of peptidergic nerve fibres appears to be large (18.5 +/- 6.6 C.U.) in proportion to the total observed area. The innervation of the drainage angle of the guinea pig eye has been well described by many authors. This is the first study to report quantitative measurements of three types of peptidergic nerve fibres identified and measured in this area. The presence of these three neurotransmitters in the trabecular meshwork of guinea pig eye suggests their possible participation in aqueous humor regulation.