Survival of Pacinian corpuscles after denervation in adult rats

Summary The ultrastructure of Pacinian corpuscles located on the crural interosseous membrane was studied in adult rats 6 h to 10 months after transection of the right sciatic nerve. Axon terminals degenerated one day after transection and were engulfed and resorbed by cells of the inner core within one week. The axial space left after removal of the axonal debris was closed by the lamellae of the inner core. The main structural features of the inner core and capsule remained preserved after denervation throughout the period of study. The denervated inner cores, however, became atrophic 10 months after neurotomy, their mean diameter being reduced by 17.5% compared with that of contralateral control corpuscles. The number of capsular lamellae was unaltered, and perineurial pathways of the peripheral nerve stump remained preserved. Schwann cells proliferated and formed Büngner bands during the first month after denervation, but retracted their processes and became atrophic at later stages after neurotomy.Survival of Pacinian corpuscles after long-term denervation in adult rats is in contrast to their rapid degeneration within several days after nerve section in neonates.     

Enhancement of nerve regeneration and orientation across a gap with a nerve graft within a vein conduit graft: a functional, stereological, and electrophysiological study

In this study, the right sciatic nerves of 40 rats were used to determine whether a nerve graft within a vein graft might accelerate and facilitate axonal regeneration, compared with a nerve graft alone. The animals were separated into four groups, as follows: group 1, sham control; group 2 (control), segmental nerve resection and no repair; group 3, segmental nerve resection and nerve grafting; group 4, segmental nerve resection and reconstruction with a nerve graft within a vein conduit graft. For all groups, sciatic functional indices were calculated before the operation and on postoperative days 7 and 90. On postoperative day 90, the sciatic nerves were reexposed and nerve conduction velocities were recorded. The sciatic nerves were harvested from all groups for counting of the myelinated axons with a stereological method. No statistically significant differences with respect to return of gait function, axon count, or nerve conduction were noted between groups 3 and 4 (p > 0.05). However, functional recovery in group 4 on postoperative day 90 was significant, compared with group 2 (p < 0.05); the recovery difference between groups 2 and 3 was not significant (p > 0.05). This study was not able to demonstrate any functional benefits with the use of a nerve graft within a vein graft, compared with standard nerve grafting.     

Retrograde degeneration within the dorsal motor vagal nucleus following bilateral vagotomy in rabbits

The effects of unilateral and bilateral intrathoracic vagotomy on the neuronal structure of the dorsal motor nucleus of the vagus were studied in rabbits. Degeneration affected mainly the small neurones which disintegrated and vanished from dorsal motor nucleus relative to the survival time after operation. A substantial proportion of large neurones was lost or degenerated, but some were preserved unchanged. In unilaterally vagotomized rabbits the dorsal motor nucleus of the intact side showed scattered neurones with axonal reaction which stands up for peripheral crossing of the vagi. The degree of retrograde degeneration was largely determined by the survival time. The nucleus ambiguus was bilaterally preserved unchanged.     

Acetylcholinesterase activity in nerve endings of tails of Rana japonica and R. catesbeiana during metamorphosis

Summary In anuran tadpole tails, the myelinated motor nerve fibers branch in the myoseptum to innervate both red and white muscle fibers at, or near, their ends. There are no significant ultrastructural differences between the nerve endings of the two types of muscle fibers.Intense acetylcholinesterase reaction product was observed in synaptic clefts and junctional folds, as well as in transverse tubules. As metamorphosis proceeded, the junctional folds of the nerve endings disappeared, however, acetylcholinesterase reaction product was still observed in the synaptic clefts. As muscle fibers began to degenerate, nerve endings began to separate from them. However, after nerve endings were completely separated from the surfaces, degenerated muscle fibers, synaptic and cored vesicles were still well preserved although no acetylcholinesterase reaction product was found. It seems clear that the mechanism of the muscle degeneration in the tadpole tail during metamorphosis is not the result of the degeneration of its nerve endings.     

Axonal protection by Nmnat3 overexpression with involvement of autophagy in optic nerve degeneration

Axonal degeneration often leads to the death of neuronal cell bodies. Previous studies demonstrated the crucial role of nicotinamide mononucleotide adenylyltransferase (Nmnat) 1, 2, and 3 in axonal protection. In this study, Nmnat3 immunoreactivity was observed inside axons in the optic nerve. Overexpression of Nmnat3 exerts axonal protection against tumor necrosis factor-induced and intraocular pressure (IOP) elevation-induced optic nerve degeneration. Immunoblot analysis showed that both p62 and microtubule-associated protein light chain 3 (LC3)-II were upregulated in the optic nerve after IOP elevation. Nmnat3 transfection decreased p62 and increased LC3-II in the optic nerve both with and without experimental glaucoma. Electron microscopy showed the existence of autophagic vacuoles in optic nerve axons in the glaucoma, glaucoma+Nmnat3 transfection, and glaucoma+rapamycin groups, although preserved myelin and microtubule structures were noted in the glaucoma+Nmnat3 transfection and glaucoma+rapamycin groups. The axonal-protective effect of Nmnat3 was inhibited by 3-methyladenine, whereas rapamycin exerted axonal protection after IOP elevation. We found that p62 was present in the mitochondria and confirmed substantial colocalization of mitochondrial Nmnat3 and p62 in starved retinal ganglion cell (RGC)-5 cells. Nmnat3 transfection decreased p62 and increased autophagic flux in RGC-5 cells. These results suggest that the axonal-protective effect of Nmnat3 may be involved in autophagy machinery, and that modulation of Nmnat3 and autophagy may lead to potential strategies against degenerative optic nerve disease.     

When should nerve gaps be grafted? An experimental study in rats

In conclusion, animal experiments have shown the following: (1) extensive elevation (mobilization) of a nerve from its bed does not interfere with its capacity to regenerate as long as the longitudinal epineural vessels are preserved, (2) suturing nerve ends under tension has a deleterious effect on the final results, (3) when a segment of nerve has been resected, the remaining nerve and the site of repair can lengthen to accommodate joint extension (within limitations), (4) if there is a segmental loss of nerve and if the nerve ends can be approximated with 10-0 epineural sutures, even if the joints must be fully flexed, the result is better than using a nerve graft, and (5) when a graft is required, it is important to avoid reversing the nerve graft. We believe direct nerve repair is preferred when flexion of the joints and mobilization of the nerve ends permits approximation with 10-0 epineural suture.     

PHOSPHOLIPASE A ACTIVITIES IN NORMAL AND SECTIONED RAT SCIATIC NERVE

Abstract— The phospholipase A₁ (EC 3.1.1.32) and A₂ (EC 3.1.1.4) activities of rat sciatic nerve homogenates have been studied. With phosphatidylcholine as substrate normal nerve had significant activity of both types at pH 5.0. Substantial increases occurred in nerve undergoing Wallerian degeneration after transection, beginning as early as 2 days after operation and rising to eight times normal values by the second week.     

Pyramidal tract function during onset of brain death

In 51 patients with primary brain lesions, who fulfilled the criteria of brain death, sequential recording of transcranial magnetic evoked potentials (TMEPs) and somatosensory evoked potentials (SEPs) were performed.In all comatose patients with apnoeic cranial nerve areflexia the TMEP could not be elicited, while the response after cervical magnetic stimulation was always preserved. Similarly, no cortical SEPs were preserved in apnoeic cranial nerve areflexia; however, the cervical somatosensory response was preserved in 44%.In deteriorating patients with coma grade III TMEPs were preserved in 3 instances, while cortical SEPs were already absent. Current brain death criteria, however, were not challenged, as TMEPs were absent in all 51 patients, at the latest when apnoea was noted.     

Free flap from the flexor aspect of the wrist for resurfacing defects of the hand and fingers

The distal portion of the flexor aspect of the forearm has been used as the donor site of full-thickness skin grafts, venous skin grafts, and Chinese forearm flaps. This article describes the use of a free flap harvested from the flexor aspect of the wrist and based on the superficial palmar branch of the radial artery to repair skin defects of the hand and fingers. The advantages of this flap are as follows: (1) the operative field is the same; (2) the radial artery is preserved; (3) it is thin, pliable, and hairless and thus can supply a gliding surface for tendons beneath it; (4) when it involves a palmaris longus tendon and/or the palmar cutaneous branch of the median nerve, it can be used as a vascularized tendon or nerve graft; and (5) in view of the flow-through type of the pedicle of the flap, the digital artery can be reconstructed simultaneously. However, it should be noted that a hypesthesia in the proximal central carpal area remains when the palmar cutaneous branch of the median nerve is harvested as a vascularized nerve graft. The scar of the donor site should be left in the distal wrist crease. If it is not lying in the distal wrist crease, it may suggest that the patient has tried to commit suicide.     

FINE STRUCTURAL LOCALIZATION OF CHOLESTEROL-1,2-3H IN DEGENERATING AND REGENERATING MOUSE SCIATIC NERVE

The localization of ³H-labeled cholesterol in nerves undergoing degeneration and regeneration was studied by radioautography at the electron microscope level. Two types of experiments were carried out: (a) Cholesterol-1,2-³H was injected intraperitoneally into suckling mice. 5 wk later, Wallerian degeneration was induced in the middle branch of the sciatic nerve, carefully preserving the collateral branches. The animals were then sacrificed at various times after the operation. During degeneration, radioactivity was found over myelin debris and fat droplets. In early stages of regeneration, radioactivity was found in myelin debris and regenerating myelin sheaths. Afterwards, radioactivity was found predominantly over the regenerated myelin sheaths. Radioactivity was also associated with the myelin sheaths of the unaltered fibers, (b) Wallerian degeneration was induced in the middle branch of the sciatic nerves of an adult mouse, preserving the collateral branches. Cholesterol-1,2-³H was injected 24 and 48 hr after the operation and the animal was sacrificed 6 wk later. Radioactivity was found in the myelin sheaths of the regenerated and unaltered fibers. The results from these experiments indicate that: (a) exogenous cholesterol incorporated into peripheral nerve during myelination remains within the nerve when it undergoes degeneration. Such cholesterol is kept in the myelin debris as an exchangeable pool from which it is reutilized for the formation of the newly regenerating fibers, especially myelin. (b) exogenous cholesterol incorporated into the nerves at the time that degeneration is beginning is also used in the formation of new myelin sheaths during regeneration, (c) mature myelin maintains its ability to incorporate cholesterol.     

The ultrastructure of the endings of nerve fibers of ordinary hairs of mammals under normal conditions and during degeneration and regeneration

At ultrastructural level, three zones are distinguished of the palisade endings of nerve fibers, most of them in contact with the basement membrane of the outer epithelial sheath of the radix pili on rat muzzle. After division of the supplying nerve (n. infraorbitalis) in test animals of the same species, its nerve fibers and palisade endings undergo degeneration, followed by regeneration within 4-5 months. Changes in the ultrastructure of the mentioned nerve elements are thoroughly studied and demonstrated in the course of the two processes. While within 6...12 days of the operation, the palisade endings degenerate, 130 days after the intervention most of them are restored and exhibit an appreciable difference relative to their typical ultrastructure in normal state. The Schwann's cells processes, enveloping degenerated and regenerated palisade endings, similarly display marked fine-structure alterations.     

Degeneration of the laminated corpuscles (of Vater--Pacini) following colchicine application to a nerve]

Laminated pacinian corpuscles from the cat mesentery have been studied morphologically and morphometrically after nerve section and colchicine application to the nerve and the results obtained are represented. Similar interventions in the nerve produce changes in the receptors resembling those of wallerian type degeneration, degeneration rate after sectioning being higher than after colchicine application. At early stages after colchicine application the internal cone and its nuclei increase in size. The data obtained suggest the nuclei of the internal cone to be under neurotrophic control of the sensory neuron that might be realized via axoplasmic transport of substances.     

Ipsilateral and cross-over elongation of the motor nerve by nerve grafting: an experimental study in sheep

In difficult reconstructions, ipsilateral or cross-over nerve grafting is sometimes necessary to achieve reinnervation and motor function. This experimental study in sheep was to answer the question of limitation of elongation of a motor nerve by grafting, the question of the optimal time for suturing the nerve graft to the muscle nerve, and the question of the successful application of this surgical technique in extremities. In 18 sheep, the vastus nerve was elongated by a saphenous nerve graft as long as possible up to 30 cm (step 1). In 10 animals the nerve graft was applied ipsilaterally, and in 8 animals it was used as a cross-over nerve graft to the contralateral limb. The time between nerve grafting and connection of the distal end of the nerve graft to the freshly cut rectus nerve supplying the rectus femoris muscle (step 2) was variable: 0, 3, 6, 9, and 12 months. In all animals, the final experiments (step 3) were performed 6 months after the last operation (step 2). Muscle force measurements in the rectus femoris muscle and quantitative analysis of the number and diameter of myelinated nerve fibers in cross sections of the nerve biopsies at different levels showed that elongation of a motor nerve by nerve grafting is principally not limited. The functional results were rather inhomogeneous and therefore unpredictable (ipsilateral group: maximum tetanic tension = 27 to 172 N; cross-over group: 0 to 227.5 N). Nevertheless, crossover nerve grafting is recommended for selected cases even in extremities. There was no correlation between the time interval between the two operations and the functional or morphologic results, although better functional results were obtained when the distal nerve suture (step 2) was performed some months after nerve grafting (step 1). A clear correlation was found only between the number of regenerated axons in the rectus nerve behind the second suture line and the muscle function.     

The effect of long-term denervation on the ultrastructure of Pacinian corpuscles in the cat

Summary The ultrastructure of Pacinian corpuscles of the cat located in the crural region and innervated by the interosseous nerve was studied 1 to 14 months after denervation. Both the Pacinian inner core and capsule remained well preserved one month after denervation. However, the denervated inner cores underwent progressive atrophy and wasting, which resulted in a gradual reduction of the amount of inner-core cells and lamellae, widening of interlamellar clefts, formation of empty spaces in the axial region and a considerable increase in the number of collagen fibrils. In spite of the wasting, the inner core still survived 14 months after denervation, but at least half of its volume became occupied by collagen fibrils which surrounded the remaining inner-core cells and lamellae. Collagen fibrils assembled in the denervated core were markedly thinner than those found in the capsule, as is also the case in normal Pacinian corpuscles. In the capsule, discrete focal degeneration, occasional pyknosis of the innermost capsular cells and macrophage infiltration were observed from the first month after nerve section onward, but the number of capsular layers remained within the normal range (30–40) up to 14 months after denervation.     

Changes of the dermal-fascial autograft after transplantation on the vascular-nervous connections using a microsurgical technique

In 56 dogs 86 microsurgical operations on transplantation of free dermal-facial autografts from the internal knee surface have been performed on the hidden vascular-nervous bundle. The animals have been observed for 1 day up to 1 year. The implanted grafts (63) have been studied, using a complex of anatomical, histological and roentgenological methods. During early time (up to 7 days) after the operation in the flap signs of edema, dystrophy and inflammatory infiltration of tissues predominate. The graft gets blood at the expense of the restored main artery and has no vascular connections with the surrounding tissues. Its nervous conductors are fragmented. During 2 weeks--1 month epidermis completely regenerates along the line of the dermal suture. In the flap bed mature granulations result in vascular connections with its surrounding tissues. These connections become stable by the end of the first month, this means that the graft has implanted. Its nervous fibers are also restored. Long-term observations demonstrate that the adaptive changes of the flap and its vascular bed are near to completion. By the end of the 1st year restoration of the main innervational connections of the graft takes place. According to the data obtained, the nervous conductors grow into it along the sewed hidden nerve and along the course of paravasal nerve plexuses. Across the scar from the surrounding tissues the dermal-fascial autograft does not reinnervate.     

NADPH-diaphorase-positive nerve cells in heterotopic spinal cord transplants

The method of ectopic transplantation of embryonic CNS rudiments makes it possible to study the mechanisms underlying adaptation of the transplanted embryonic rudiments. The production of nitric oxide by cells is considered as one of such mechanisms. NADPH-diaphorase is an index of the presence of nitric oxide synthase in cells. It was shown that the nerve cells of rat embryonic spinal cord transplants preserved their capacity to express NADPH-diaphorase after transplantation in the sciatic nerve of an adult animal for six months. The dynamics of NADPH-diaphorase-positive neurons of rat embryonic spinal cord developing after transplantation and in situ were studied. In spinal cord neck region, small bipolar NADPH-diaphorase-positive neurons were visualized on day 17 of prenatal development. After transplantation of the embryonic (day 15) spinal cord in the nerve, NADPH-diaphorase-positive neurons were formed later than in situ: within seven days. The results of histochemical studies carried out within six months after the operation suggest a protective role of NADPH-diaphorase in the neurons of allotransplants developing under the conditions of altered microenvironment and insufficient innervation and also suggest that nitric oxide can cause the death of neurons in long surviving transplants.     

Use of an innervated skin graft to provide sensation to the reconstructed heel.

An innervated skin graft--a full-thickness skin graft with its nerve supply intact--is presented. In the case described, the sural nerve was preserved and an island of skin supplied by it was transposed to provide sensation to a heel pad, which had first been reconstructed by a cross-thigh flap. This transposed sensation has allowed full activity for 3 years in a young boy wearing normal footwear. This suggests that ulceration of all, or most, heel reconstructions is due to lack of sensation and, further, that the necessary sensation can be provided by this technically simple transposition of an innervated skin graft.     

Glial and Neuronal Marker Proteins in the Silicone Chamber Model for Nerve Regeneration

Abstract: In the present study, neuronal and Schwann cell marker proteins were used to biochemically characterize the spatiotemporal progress of degeneration/regeneration in the silicone chamber model for nerve regeneration. Rat sciatic nerves were transected and the proximal and distal stumps were inserted into a bridging silicone chamber with a 10-mm interstump gap. Using dot immunobinding assays, S-100 protein and neuronal intermediate filament polypeptides were measured in different parts of the nerve 0–30 days after transaction. In the most proximal nerve segment, all the measured proteins were transiently increased. In the proximal and distal stumps adjacent to the transection, the studied proteins were decreased indicating degeneration of the nerve. Within the silicone chamber, the regenerating nerve expressed the Schwann cell S-100 protein already at 7 days, whereas the neurofilament polypeptides appeared later. These observations are corroborated by previous morphological studies. The biochemical method described provides a new and fast approach to the study of nerve regeneration.     

Mast cell distribution and density in Lewis rat gingiva after bilateral neurectomy of inferior alveolar nerves

The effect of uni- and bilateral neurectomy of inferior alveolar nerve on mast cell (MC) density and topography were studied in Lewis rat gingival mucosa. The results were compared with unilateral sham operation effects. The samples were taken in each group both form left and right side of gingiva, MC were revealed by pinacyanol erythrosinate and counted in connective tissue adjacent to the basal membrane of gingival epithelium. MC were insensitive to sham operation and the differences were noted only in MC density after unilateral neurectomy between both sides of gingiva, and in distribution between the control and left side after bilateral neurectomy. The results suggests that neurectomy and sham operation trauma after 3 weeks have limited effect on mast cell density and distribution.     

NADPH-Diaphorase-Positive Nerve Cells in Heterotopic Spinal Cord Transplants

The method of ectopic transplantation of embryonic CNS rudiments makes it possible to study the mechanisms underlying adaptation of the transplanted embryonic rudiments. The production of nitric oxide by cells is considered as one of such mechanisms. NADPH-diaphorase is an index of the presence of nitric oxide synthase in cells. It was shown that the nerve cells of rat embryonic spinal cord transplants preserved their capacity to express NADPH-diaphorase after transplantation in the sciatic nerve of an adult animal for six months. The dynamics of NADPH-diaphorase-positive neurons of rat embryonic spinal cord developing after transplantation and in situwere studied. In spinal cord neck region, small bipolar NADPH-diaphorase-positive neurons were visualized on day 17 of prenatal development. After transplantation of the embryonic (day 15) spinal cord in the nerve, NADPH-diaphorase-positive neurons were formed later than in situ: within seven days. The results of histochemical studies carried out within six months after the operation suggest a protective role of NADPH-diaphorase in the neurons of allotransplants developing under the conditions of altered microenvironment and insufficient innervation and also suggest that nitric oxide can cause the death of neurons in long surviving transplants.