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Incorporation of glycoproteins into peripheral nerve myelin.

Peripheral nerve myelin contains a dominant low molecular weight glycoprotein called Po. To study the metabolism of this glycoprotein, tritiated fucose was injected into the peripheral nerves of adult mice and developing rats, and the temporal distribution of label was examined by autoradiography and gel electrophoresis. Mice and rat pups, injected with fucose, were sacrificed from 1 h to 98 days later. Series of autoradiographs were prepared. At the shortest labeling periods, newly formed product was confined to juxtanuclear Schwann cell cytoplasm, in association with regions rich in Golgi apparatus. After longer labeling periods, silver grain levels in Schwann cell cytoplasm decreased; concomitantly, there was an increase of silver grains associated with myelin. In adult animals, label associated with myelin was concentrated over outer layers of thickly myelinated fibers. Even at the longest time intervals examined (72 and 98 days), this distribution of label was largely retained. In contrast, in developing animals, label became associated with inner layers of the thicker sheaths. At no time was label observed over axons. Gel electrophoresis revealed that tritiated fucose was a suitable precursor for the faster migrating peripheral nerve glycoprotein(s). At all times examined, there was a single major peak of radioactivity that co-migrated on sodium dodecyl sulfate (SDS) acrylamide gels with the Po protein. Sometimes, a faster migrating shoulder of radioactivity was noted. With increased labeling periods, there was an enrichment of radioactivity associated with Po, indicative of a relatively slow turnover rate.

Animals

Recent developments in peripheral nerve surgery. Management of open traumatic peripheral nerve lesions.

The modern technique of operative repair of traumatic nerve interruption is discussed. The results of microsurgical repair have proved to be superior to those of conventional methods. Besides the anatomy and the operative technique, the indications and timing for surgical intervention are discussed. Reference to the literature and the authors' own experience show that delayed primary repair is the method of choice.

Humans

Subperineurial space of the sural nerve in various peripheral nerve diseases.

The size of the subperineurial space of the sural nerve has been evaluated quantitatively in 69 cases of various peripheral nerve diseases and in controls. A significant increase was found in beriberi neuropathy (6 cases) and idiopathic polyradiculoneuropathy (9 cases) as compared with the control (8 cases). On electron microscopy a few macrophages, fibroblast processes, collagen fibrils with a diameter of 50 nm, microfibrils with a diameter of 8 nm, and amorphous material were observed in both the enlarged subperineural space and the endoneurial intercellular space. They were less frequently observed in controls. No significant correlation was found between the size of subperineurial space and the density of myelinated fibers.

Adolescent

Transcutaneous electrical nerve stimulation in chronic pain after peripheral nerve injury.

Transcutaneous electrical stimulation was tested in 24 patients with chronic pain following a peripheral nerve injury in an extremity, in 10 patients with a good effect. All of these 10 patients displayed signs of increased sympathetic activity in addition to hyperalgesia. Sympathetic block gave complete freedeom from pain. In 14 patients with the same symptomatology but without an increased or with only very slightly increased sympathetic activity, no or an insignificant effect was obtained. Sympathetic block did not relieve the pain in this group. Transcutaneous electrical stimulation should be tried as an alternative to sympathectomy in causalgia major or minor.

Adult

The characterization of type I and type III collagens from human peripheral nerve.

The normal chemical features of peripheral nerve collagens were determined on postmortem, histologically normal adult human femoral nerve. 1. Genetically distinct type I, [alpha1(I)2]alpha2, and type III, [alpha1(III)]3, were isolated by differential salt precipitation and the component subunit chains, alphal(I), alpha2 and alphal(III) were obtained by ion-exchange chromatography and gel filtration. 2. The molecular weight of alphal(I) and alpha2 of type I collagen was 95 000 and that for type III was 280 000. Reduction of type III with dithiothreitol yielded expected alpha1(III) chains of 95 000 molecular weight. 3. The amino acid composition of the three collagen chains, alpha1(I), alpha2, and alpha1(III), was the same as previously reported values for the corresponding chains from human skin except for slightly elevated hydroxylysine content. 4. Peripheral nerve collagen was found to contain 81% type I collagen and 19% type III. These results indicate that peripheral nerve collagen characteristics closely simulate that of human skin and differ from that of human aorta and other parenchymal organs. These data will permit a chemical analysis for possible abnormalities of peripheral nerve collagen in various neurogenic disorders.

Aged

Studies on myelin proteins in human peripheral nerve.

The myelin fraction from human peripheral nerve was prepared. Two basic protein fractions (BF-P2 and PB) were isolated from acid extracts of the myelin fraction and three glycoproteins (BR-PO, PASII and Y protein) were purified from its acid-insoluble residue. In biochemical analysis the human BF-P2 protein (M.W.13,000) showed similar but not identical properties to bovine BF-P2 protein. The PB fraction was suggested to include the encephalitogenic CNS-BP (M.W.18,000) and another, new protein of similar molecular weight. Both the human BR-PO protein (M.W.28,000) and PASII protein (M.W.13,000) showed similar biochemical properties to the corresponding myelin proteins of bovine peripheral nerve, while they both are clearly different from other myelin proteins. Close relationship between the BR-PO protein and the Y protein (M.W.22,000) was suggested by amino acid analysis. Injection of the myelin fraction of bovine peripheral nerve with the complete adjuvant produced EAN while the CNS-BP induced EAE in laboratory animals. However, all three purified proteins, BF-P2, BR-PO and PASII, from bovine peripheral nerve myelin were inactive in inducing demyelinating diseases.

Amino Acids

Differential atrophy of sensory and motor fibers following section of cat peripheral nerves.

Differential effects of peripheral nerve section on myelinated sensory and motor fiber populations were investigated in 5 hindlimb nerves of cats. Upon electrical stimulation of each nerve, monophasic compound action potentials were recorded from the L6, L7 and S1 dorsal and ventral roots, and the impedance of each root was measured. The decline in the electrical charge computed from potentials 43 to 252 days after nerve section gave a measure of the effect of axotomy on the diameters of sensory and of motor fibers in each nerve. No significant difference in the rate of atrophy of sensory and motor fibers was observed after about 45 days following nerve section. After about 145 and 245 days, however, dorsal root charge contributions had decreased significantly more than ventral root values. Exponential decay curves were fitted separately to charge data for sensory and for motor fibers. The calculated value for the endpoint of the decay was about 35% of the control value for motor fibers, and not significantly different from zero to sensory fibers. These results suggest that in response to axotomy, myelinated motor fiber diameters decline at first but later stabilize, while myelinated sensory fibers continue to decline and may atrophy completely if regeneration is prevented. Possible roles of electrical activity and of 'trophic' interactions with the periphery in the maintenance of cell properties are discussed.

Animals

Microsurgery of the peripheral nerve: a physiological approach.

Results with peripheral nerve injuries, which today still fall short of our goals, should intensify our efforts in this specialized field of microsurgery. We are living in an exciting era of functional reconstitution. Long gone are the times when a pink replanted digit or a successful free flap comprised the end-point of our surgical efforts. The direction from now on is toward achieving functional restoration by utilizing basic principles while armed with the unlimited potentials of reconstructive microsurgery. To this end we need the assistance and cooperation of other sciences since the enigma of the peripheral nerve can be deciphered only by a multidisciplinary approach.

Electrophysiology

An explanation of axonal regeneration in peripheral nerves and its failure in the central nervous system.

Nerve fibres severed within peripheral nerves are able to regenerate and reinnervate the structures they formerly supplied. Most axons severed within the mammalian central nervous system (CNS) do not regenerate in this way. Regenerative axonal growth begins to occur in the CNS but ceases about two weeks after injury. Five earlier theories purporting to explain this difference are reviewed and found not to account satisfactorily for many experimental observations. A new hypothesis is advanced in which it is held that in order for regeneration to take place, the growing tips of the axons must be surrounded by extracellular fluid containing proteins (of specified identity) derived from the blood plasma. Such proteins are thought to be imbibed by the tips of the fibres and transported retrogradely to the neuronal cell-bodies. With this hypothesis it is possible to explain the success of axonal regeneration in peripheral nerves and its failure in the CNS. It is also possible to account for the exceptional circumstances in which axons do regenerate in the CNS. Various experiments are suggested for testing the validity of the new hypothesis.

Amphibians

Tension and creep phenomena in peripheral nerve.

Tension introduced into peripheral nerves during their surgical repair may reduce the success of this procedure. Two mechanical factors are important; the tension required to effect a repair, and the rate at which this tension changes after surgery. These two factors have been investigated in the rat sciatic nerve. The results show an increasing resistance to elongation of the nerves with increasing tension. Under a constant elongation the tension in the nerves reduces by about 30 per cent in the first 10 minutes and by a small amount in the following 20 minutes.

Animals

A comparative study of the polypeptides of mammalian peripheral nerve myelin.

Polypeptide components of peripheral nerve myelin prepared from sciatic nerves of mouse, rat, rabbit, cat, guinea pig, monkey, dog and chicken were analyzed using a 20% polyacrylamide slab-gel electrophoresis system. Using this technique, we have demonstrated that there are at least two polypeptides present in the region of P1 protein (mol. wt. 18,000 and 16,000), and two closely moving components in the region of P2 protein (mol. wt. 13,500). A polypeptide (mol. wt. 10,500) is present in significant amounts only in mouse sciatic nerve myelin. Staining with periodic acid-Schiff reagent shows that in addition to P0 protein (mol. wt. 28,000), the major glycoprotein of peripheral nerve myelin, there are several minor glycoprotein components in both the high and low molecular weight regions.

Animals

Complement receptors in human peripheral nerve tissue.

Cryostat sections of human peripheral nerve tissue adsorbed sheep erythrocytes sensitized with antibody and human complement (EAC). Activated complement was essential for the reaction to occur. The receptor had specificity for the C3b fragment. In order to obrain binding to nerve tissue, the indicator cells had to be coated with more complement than that required for binding to C3b receptors in spleen and renal tissue. The receptors are located within the nerve fascicles and are probably of glycoprotein nature. The receptors for C3b in peripheral nerve tissus may be of significance in the deposition of immune complexes, which may play a role in acute polyradiculoneuritis.

Complement Activation

Characterization of the calcium-induced disruption of neurofilaments in rat peripheral nerve.

Transverse frozen sections of desheathed rat peripheral nerve were incubated in media of different composition prior to fixation and processing for electron microscopic examination. Neurofilaments remained intact when these tissues were incubated in calcium-free media. A loss of neurofilaments and their replacement by granular debris occurred in myelinated and unmyelinated fibers following incubation in media containing 2 mM calcium. The calcium-mediated disruption of neurofilaments was inhibited by preincubation or incubation with 1 mM p-chloromercuribenzoate (PCMB). The inhibition by preincubation with PCMB could be partially reversed by subsequent preincubation with 10 mM dithioerythritol (DTE). Calcium-mediated breakdown of neurofilaments did not occur after prolonged preincubation in calcium-free media, a finding which suggested that neurofilament disruption was dependent upon a tissue factor which could be lost or inactivated in frozen-sectioned nerve tissues. The findings of the present study provide morphological evidence that neurofilament disruption in mammalian peripheral nerve is mediated by a calcium-activated, PCMB-sensitive enzyme in the axoplasm of myelinated and unmyelinated nerve fibers.

Animals

Ischaemia of peripheral nerve and muscle.

Although skeletal muscle and peripheral nerves are both resistant to ischaemia there are nevertheless many syndromes in which they are affected, either separately or together. It is frequently difficult to distinguish the effects of arterial ischaemia from those of compression, which may operate through vascular occlusion, or, in the case of peripheral nerve, by mechanical deformation of nerve fibres. A great deal has been learned from experimental models, but not all of it is applicable to the complexity of human neuromuscular ischaemia which requires further study.

Arteriosclerosis

On basic proteins in bovine peripheral nerve myelin.

1. Myeline proteins in bovine peripheral nerve migrated as two main band-(BF and BR protein) and one faint middle band (BM protein) on sodium dodecyls sulfate-polyacrylamide gel electrophoresis. The relative mobility of these two main bands differed from those of myelin proteins in the central nervous system. 2. The acid extract of the myelin fraction from bovine peripheral nerve was separated into one main peak and two minor peaks on a Sephadex G-75 column. The major component of the second minor peak was the BM protein; the major component of the main peak was the BF protein. The BR protein was not extractable by acid solution. 3. Molecular weights of the BF, the BM and the BR protein were determined as around 13 000, 20 000 and 28 000, respectively, by sodium dodecylsulfate-polyacrylamide gel electrophoresis. 4. The amino acid composition of the BF protein was quite different from the encephalitogenic protein and the Folch-Lees type proteolipid protein in the central nervous system. However the BM protein showed similar amino acid composition to the encephalitogenic protein. 5. The tryptic peptide maps of the BF protein and of the encephalitogenic protein were quite different. The results suggested that the amino acid sequences of these two proteins are different and that they contain no common tryptophan-containing peptide.

Amino Acid Sequence

Peripheral nerve abnormalities related to galactose administration in rats.

Electrophysiological, biochemical, and morphometric observations were made on the peripheral nerves of rats after galactose feeding. Motor nerve conduction velocity was found to be reduced. This was associated with an accumulation of galactitol in the peripheral nerves and a diminution in their myoinositol content. An increased water content and fascicular area, taken in conjunction with a probable increase in the area of the endoneurial spaces, indicated overhydration of the peripheral nerves. Morphometric observations on the myelinated fibre population in the tibial nerve showed no loss of fibres and although both the maximal and the average diameter of the myelinated fibres was slightly less than in age-matched controls, this was insufficient to explain the reduction in conduction velocity. Segmental demyelination was not detected and the relationship between myelin thickness and axon circumference was not altered. Electron microscope observations revealed no ultrastructural changes in the myelinated fibres and, in particular, no abnormalities at the nodes of Ranvier or indication of abnormal hydration of the Schwann cells. The relevance of these findings to the peripheral nerve changes in human and experimental diabetes is discussed.

Animals

Somatic unmyelinated fibre degeneration in rabbits with granulomatous hypersensitivity produced by a non-myelin antigen in sensory peripheral nerve.

Rabbits previously sensitised by injection of human sensory peripheral nerve showed degeneration of somatic unmyelinated fibres at skin test sites when challenged by sensory nerve or certain non-myelin fractions prepared from human sensory nerve. Epithelioid cells were found in the endoneurium and may mediate the nerve damage. The degeneration was consistently found and lasted for at least 58 days. Phagocytosing macrophages were absent. Differences in the pattern of nerve damage to that of EAN and EAE are discussed.

Animals