Distribution of P0 protein and the myelin-associated glycoprotein in peripheral nerves from Trembler mice.

Summary The Trembler mouse has a dysymelination of peripheral nerves that includes hypomyelination, failure of myelin compaction, and demyelination/remyelination. We have localized the myelin proteins P₀ and myelin associated glycoprotein in Trembler peripheral nerve and correlated their distributions with the ultrastructure of myelin internodes. Immunocytochemically, myelin-associated glycoprotein was localized in Schwann cell periaxonal membranes, Schmidt-Lanterman incisures, paranodal loops, and internal and external mesaxons. P₀ staining was located over compact myelin and regions of Schwann cell cytoplasm rich in Golgi membranes. An unusual abundance of small, P₀-stained, Golgi-related vesicles was found in some Schwann cells. P₀ protein was also detected in multiple spiral wraps of myelin-associated glycoprotein-positive mesaxon membranes. At some sites the periodicity of the myelin membranes was intermediate to that found in mesaxon membranes and compact myelin. The distance between apposing extracellular leaflets was similar to that found in mesaxon membranes, while the cytoplasmic leaflets were fused but twice as thick as normal major dense lines. These intermediate membranes were stained by P₀ and myelin-associated glycoprotein antiserum. These studies suggest that altered transport and/or translocation of P₀ and myelin-associated glycoprotein results in defective myelin compaction in Trembler peripheral nerve.     

Electron microscopic demonstration of polysaccharides in central and peripheral myelin by thiosemicarbazide-protein-silver staining

Summary Thin sections of glutaraldehyde-fixed central and peripheral nerve myelin were stained with thiosemicarbazide and protein-silver after oxidation with periodic acid on thin sections. In compact CNS myelin, staining was observed exclusively on intraperiod lines. In peripheral myelin, both intraperiod and major dense lines were stained. In addition, dense staining was observed on plasma membranes of oligodendrocytes and Schwann cells, especially periaxonally on tongue processes and in Schmidt-Lanterman incisures. The observed staining was most prominent on glycogen granules in unfixed and freeze-substituted tissues. Therefore, the results strongly suggest that polysaccharides of glycoproteins and glycolipids are visualized in both CNS and PNS compact myelin as well as on surface membranes of oligodendrocytes and Schwann cells.     

Localizations of ruthenium red positive material in rabbit peripheral nerves

The penetration and distribution of ruthenium red in the axon-myelin-Schwann cell complex of developing rabbit peripheral nerve fibers are investigated. Ruthenium red positive material is established in the axoplasm, axolemma, periaxonal space, major dense lines and intraperiod lines of the compact myelin, mesaxons, split peripheral myelin lamellae, Schmidt-Lanterman and longitudinal incisures, paranodal loops and axo-glial contacts, Schwann cell cytoplasm and basal lamina, nodal extracellular matrix, desmosome-like structures, endoneural collagen. Some features of the distribution of the contrast material in the developing myelin sheath are described. Regional differences of the axolemma and of the Schwann cell cytoplasm and plasmalemma are established. The prevalence of glycoproteins or glycolipids in the ruthenium red stained material in its different localizations is discussed on the basis of trypsin and hyaluronidase digestion performed.     

Ultrastructural cytochemical localization of 5'-nucleotidase activity in axon-myelin-Schwann cell complex

5'-Nucleotidase activity has been localized at the ultrastructural level in the axon-myelin-Schwann cell complex. Sciatic nerves of rabbits of pre- and postnatal development were used. Positive reaction was found on the plasma membrane, basal lamina, cytoplasm, and finger-like processes of the Schwann cells; on the intraperiod lines of the compact myelin, on the surface of myelin sheath, in the split myelin lamellae in the paranodal regions and Schmidt-Lanterman clefts, in segments of outermost and innermost lamellae, split off from the interparanodal myelin, in the mesaxons (outer and inner), in the loose myelin lamellae in the earlier stages of myelinization; on the axolemma (especially in the nodal and paranodal segments), in the periaxonal space, axoplasm. The alterations of 5'-nucleotidase distribution were associated with the developing myelin sheath.     

Freeze-etching images of central myelinated nerve fibres

Summary The three-dimensional features of myelinated nerve fibres in rat brain were studied with the freeze-etching technique. The cross fracture of the myelin sheath exhibited an alternate orientation of ridges and furrows with a periodicity of about 60–70 Å. The ridge in the myelin sheath was either continuous with the outer ridge of the outer loop plasma membrane or formed by fusion of the inner ridge of the loop membrane. The furrow in the myelin sheath was continuous with the furrow of the loop plasma membrane. Since the ridge of the loop membrane practically corresponded to the leaflet of the loop unit membrane, the ridge in the myelin sheath was roughly equivalent to the major or the intraperiod dense line, and the furrow in the myelin sheath was then localized in the middle between the major and the intraperiod dense lines. The myelin sheath in tangential fractures was composed of repeating membrane layers, which were relatively free of membrane particles. Although the furrow in cross fractures generally corresponded to the potential fracture plane, the main fracture process in the myelin sheath often passed alternately in the potential fracture plane. The fracture face of the myelin sheath, therefore, might represent an extended lipid layer localized in the middle of the lipid bilayer.     

Progressive axonopathy: An inherited neuropathy of boxer dogs. 4. myelin sheath and Schwann cell changes in the nerve roots

Summary Changes in the myelin sheath have been studied in the nerve roots of dogs with Progressive axonopathy, an autosomal recessive inherited neuropathy. The earliest changes were attenuation of the sheath at the proximal paranode and adjacent internode, probably in response to the axonal swelling which occurs in this area. Myelin bubbles were frequently observed along internodes. As the disease developed, progressively more fibres demonstrated short internodes of irregular length and thin myelin sheaths suggesting extensive remyelination and remodelling of the sheath. Short lengths of axons devoid of myelin, and occasional macrophages were also encountered. Sheaths of both original and newly formed internodes were highly irregular in outline. Occasional intra-axonal projections of adaxonal Schwann cell cytoplasm were observed, but complex interdigitations were unusual.A moderately electron-dense, granular material accumulated within the myelin sheath, becoming more obvious in the advanced disease. This material of unknown origin and composition was located predominantly at the intraperiod line principally between the adaxonal cytoplasm and the inner major dense line, but also at Schmidt-Lanterman incisures and between paranodal loops. Xenografts of the canine nerves into athymic mice failed to demonstrate any of the myelin sheath changes. The temporal and spatial relationship of the myelin sheath and axonal changes and the failure to reproduce the natural lesion in grafts suggest that Schwann cell alterations probably occur in response to the axonal changes.     

Segmental demyelination in peripheral nerves of old cats

This study of the fine structure of sciatic nerve branches in normal old cats provides evidence indicating that segmental demyelination may account, in part, for the significant decrease with age in the mean axonal conduction velocity in these hindlimb nerves. Fibers of different diameters exhibited focal abnormalities of their myelin sheath. Lipid-like droplets and granulo-vacuolar debris were present in distended portions of the inner adaxonal rim and in the outer cytoplasmic compartment of the Schwann cell. These inclusions extended into the cytoplasm of the paranodal myelin loops and clefts of Schmidt-Lantermann. There also occurred disruption of the axoglial junctions and separation of the myelin loops from the paranodal axolemma which widens the nodes of Ranvier. Complete disruption of one or more contiguous segments of the myelin sheath was produced by interlamellar splitting and ballooning along the major dense and intraperiod lines. Axonal degeneration occurred less frequently and was not present in all hindlimb nerves.     

Development of early postnatal peripheral nerve abnormalities in Trembler-J and PMP22 transgenic mice

Mutations in the gene for peripheral myelin protein 22 ( PMP22 ) are associated with peripheral neuropathy in mice and humans. Although PMP22 is strongly expressed in peripheral nerves and is localised largely to the myelin sheath, a dual role has been suggested as 2 differentially expressed promoters have been found. In this study we compared the initial stages of postnatal development in transgenic mouse models which have, in addition to the murine pmp22 gene, 7 (C22) and 4 (C61) copies of the human PMP22 gene and in homozygous and heterozygous Trembler-J ( Tr ^J ) mice, which have a point mutation in the pmp22 gene. The number of axons that were singly ensheathed by Schwann cells was the same in all groups indicating that PMP22 does not function in the initial ensheathment and separation of axons. At both P4 and P12 all mutants had an increased proportion of fibres that were incompletely surrounded by Schwann cell cytoplasm indicating that this step is disrupted in PMP22 mutants. C22 and homozygous Tr ^J animals could be distinguished by differences in the Schwann cell morphology at the initiation of myelination. In homozygous Tr ^J animals the Schwann cell cytoplasm had failed to make a full turn around the axon whereas in the C22 strain most fibres had formed a mesaxon. It is concluded that PMP22 functions in the initiation of myelination and probably involves the ensheathment of the axon by the Schwann cell, and the extension of this cell along the axon. Abnormalities may result from a failure of differentiation but more probably from defective interactions between the axon and the Schwann cell.     

Wallerian degeneration: A reevaluation based on transected and colchicine-poisoned nerves in the amphibian,Triturus

The sequence of degeneration in colchicine-poisoned or in transected peripheral nerve fibers is reported. The results lead us to conclude that the Schwann cell is the predominant agent of axon and myelin destruction. The Schwann cytoplasm burgeons, swells the Schmidt-Lanterman clefts and the adaxonal layer of cytoplasm, and invades the electron-dense bands of myelin. The axolemma and adaxonal membranes are eroded and Schwann organelles invade the axoplasm commingling with those of the axon to form a peripheral “reactive zone.” The reactive zone of organelles surrounds a core of compacted neurofilaments which may persist for days. Eventually the entire contents, commingled organelles and neurofilaments, are destroyed. The myelin sheath is destroyed without being separated from its Schwann cell of origin.     

Double myelination of axons in the sympathetic nervous system

Summary Relationships between axons and Schwann cells in myelinated fibres of the superior cervical (sympathetic) ganglion have been examined in normal adult rats. In cross-sections through the ganglion up to 4 % of myelinated fibres were focally encircled by an additional myelinating Schwann cell, forming regions termed double myelination. In these regions and elsewhere in the ganglion, the structure of the inner fibre (axon and myelinating Schwann cell) conformed to the relationships expected on the basis of numerous previous investigations on normal peripheral nerve. However, the outer Schwann cell and myelin sheath, which formed an annulus around the inner fibre, was remarkable in that it apparently made no direct contact either with the centrally enclosed axon or with any neighbouring axon, yet appeared largely if not completely intact. In addition, the increasing frequency of double myelination in older animals and the rarity of myelin degeneration in the same ganglia indicate that the outer Schwann cell, and in particular its myelin sheath, persist for some period in an isolated form. Double myelination was not located in non-sympathetic peripheral nerve samples from the same animals. Double myelination may result from the displacement of one myelin internode by the interposition of another Schwann cell rendering the original Schwann cell redundant. There was no involvement of haematogenous cells as occurs in some demyelinating conditions. While some parallels may be found with previous studies, this would appear to be the first report of apparent survival of myelin in a Schwann cell not making, as far as could be determined in the present study, at least partial direct axonal contact. These observations on sympathetic nerve may provide a new perspective on axon-Schwann cell signalling.     

Myelin intrusions in beaded nerve fibers.

Small intrusions form in the internodes in or near the constrictions of beaded fibers prepared by fast-freezing and freeze-substituting mildly stretched nerves in the cat and rat. They appear as inwardly directed folds of the inner lamellae of the myelin sheath, or regularly formed spheres composed of lamellae with major dense and interperiod lines like those of the myelin sheath. A splitting of the lamellae and separation of the major dense lines may occur with an accumulation of Schwann cell cytoplasm between them, the result of an influx of cytoplasmic fluid from nearby constrictions. Longitudinally oriented microtubules have been observed in the intrusions, in the adaxonal Schwann cell cytoplasm, and in the innermost lamellae of the myelin sheath. The paranodes contain a number of larger intrusions in the form of spurs and globules along with shelve-like folds of the myelin sheath oriented in the longitudinal direction. Axoplasmic fluid driven from the constrictions during beading can enter the paranodes to smooth out their folds leaving the globular and spur-shaped myelin intrusions in isolation. Their wall thickness, measured from the central opening to the surface of the intrusion, is the same as that of the myelin sheath or, in some cases, double, the result of the folding of a spur-like intrusion upon itself. Intrusions unconnected to the sheath are seen in unbeaded fibers with regular, compact lamellae surrounded by axolemma. Others lack a covering axolemma and consist of variably disorganized and irregularly shaped lamellae suggesting that they are undergoing fragmentation and dissolution within the axon. The hypothesis is advanced that the intrusions in the internodes arise from an excess of lipid and other myelin components when the diameter of the sheath is reduced in the beading constrictions. In the paranodes, excess myelin components moved into these regions form the shelf-like folds which may fuse to form intrusions. These, separated from the myelin sheath, undergo fragmentation and dissolution and are carried by retrograde transport to the cell bodies where their constituent components can be reutilized.     

Distribution of Schwann cell cytoplasm and plasmalemmal vesicles (caveolae) in peripheral myelin sheaths. An electron microscopic study with thin sections and freeze-fracturing

Summary The mode of distribution of Schwann cell cytoplasm in the various regions of the peripheral myelin sheath is reviewed using freeze-fractured and thin sectioned nerves from bullfrogs, chickens and cats, perfused with an aldehyde mixture or fixedin situ with potassium permanganate.It is shown that the peripheral myelin sheath consists of two domains: one semi-compact at the outer and the inner turns, and one compact, extending from the second to the next-to-last turns. In the semi-compact domain nomajor dense line is formed, although the cytoplasmic layer may be as thin as 50 Å. As is well known, amajor dense line is formed instead in the compact domain. The whole sheath is bordered by a marginal cytoplasmic belt. This communicates with a cytoplasmic reticulum present in the semi-compact domains and with a reticulum (formed by longitudinal and circumferential incisures) in the compact domain.Many cytoplasmic channels in the semi-compact domain end blindly, as do many of the longitudinal and the circumferential incisures. It is speculated that these cytoplasmic networks are in a dynamic state, and that the changes within the semi-compact domain may be faster than those in the compact region.Numerous uncoated vesicles, 600–1100 Å in diameter, open onto the plasmalemma in the perinuclear region, along the outer mesaxon, and in the larger cytoplasmic trabeculae on the outer surface of the sheath. These plasmalemmal vesicles (caveolae) do not occur on the thinner cytoplasmic channels, on the inner surface of the sheath or on the incisures. Horseradish peroxidase studies indicate that the plasmalemmal vesicles become filled with the enzyme but do not migrate into the cytoplasm within a 4 h period. The movement of cytoplasm within the myelin sheath and the significance of the vesicles are discussed.The matter is important, since there are indications fromin vitro studies of cultured nervous tissue and of teased fibres, that movements take place in the myelin sheath. These are presumably related to cytoplasmic translocations capable of producing outpocketings and invaginations of the sheath's contour and may also involve the Schmidt—Lanterman incisures (Murray and Hermann, 1965; Singer and Bryant, 1969; Gitlin and Singer, 1974). Althoughin vivo observation of intact nerves for several hours did not reveal conspicuous movements (Williams and Hall, 1970, 1971), it is an attractive hypothesis that cytoplasmic movements are involved both in the formation and maintenance of the myelin sheath and in pathologic alterations (Robertson, 1958; Webster, 1965, 1971).On sabbatical leave from the University of TromsØ Medical School.     

Double myelination of axons in the sympathetic nervous system of the mouse. II. Mechanisms of formation

Summary The phenomenon termed double myelination, present in sympathetic nerve of normal adult rats and mice, comprises regions of a myelinated axon which are concentrically ensheathed by additional (outer) myelinating Schwann cells. Evidence has been presented that in some instances the outer Schwann cell fails to make contact with an axon, yet its myelin sheath characteristically remains ultrastructurally intact. The present study has sought to identify and analyse configurations intermediate between single and double myelination, in order to determine the mechanism(s) underlying the formation of double ensheathment. Superior cervical ganglia from normal male mice aged 12–24 months were prepared for electron microscopy by systemic aldehyde perfusion. Regions of interest were extensively serial-sectioned for detailed electron microscopical analysis and reconstruction. The earliest evidence for alteration to the expected intimate ensheathment of axons by myelinating Schwann cells involved invasion of supernumerary Schwann cells and their processes at the node of Ranvier, resulting in displacement of the paranodal pockets from axonal contact. Similar paranodal displacement occurred at heminodes as a result of lateral extension and invasion of processes from the adjacent Schwann cell (i.e. the cell investing the unmyelinated domain of the axon). Subsequently, processes of the invading cell extended progressively into internodal regions, located at all times between the plasma membranes of the axon and displaced Schwann cell. The cytoplasmic pockets at the remaining paranode were then subject to invasion. At various stages of displacement myelin formation commenced within the invading cell, representing the first acquisition of double myelin ensheathment in the development of the configuration. Involvement of haematogenous cells in displacement was not detected. There was also evidence consistent with paranodal displacement by adjacent pre-existing myelinating cells, but this additional mechanism appeared minor relative to the involvement of (initially) non-myelinating Schwann cells. We found no evidence for the alternative possibility that Schwann cells could synthesize a myelin sheath around a pre-existing myelinated axonde novo, independent of any direct axonal contact. These results are consistent with the well-established requirement for axonal contact by Schwann cells engaging in initial myelin formation, in the sense that the myelin sheath of the outer cell was synthesized prior to its displacement, and that a myelin sheath was not formed by the invading cell until it had invested the axon in a 1:1 relationship. In addition, the emergence of several key features of double myelination (infolding and continuing integrity of the outer sheath, sites of presence/absence of basal lamina on the outer cell) further supports the view that double myelination represents a culmination of these developmental stages.     

Acute stages of batrachotoxin-induced neuropathy: a morphologic study of a sodium-channel toxin

Summary The acute effects of batrachotoxin, a steroidal neurotoxin which opens the membrane sodium channel, were observed morphologically at various time points up to 3 h after injection into rat peroneal nerve. Three changes were found. First, there was massive swelling of the axon at the node of Ranvier accompanied by retraction of paranodal myelin. Second, a similar swelling of unmyelinated axons was seen. Third, extracellular fluid accumulated along the internode in the adaxonal space, the intraperiod line of myelin and, rarely, the external mesaxon, with concomitant shrinkage of the axon. The first two changes might be explained on the basis of massive shift of sodium through the batrachotoxin-modified sodium channel into the axon and subsequent osmotic shift of fluid. The reason for the third change is not clear but probably also has a ionic basis.     

Freeze-fracture analysis of myelin membrane changes in Wallerian degeneration

Summary Transection of mouse sciatic nerves produced microscopic changes in the myelin sheaths distal to the transection. Studied with freeze-fracture, these microscopic changes were correlated with alterations in the macromolecular organization of nerve membranes. In control mice, sciatic nerve myelin contained randomly distributed intramembranous particles. In the early stages of myelin breakdown the lamellae split and large areas of myelin membrane lacked intramembranous particles. The remaining particles clustered with a greater than normal density. Degenerating myelin was found within Schwann cells which still had an outer mesaxon and a normal distribution of intramembranous particles on the cell outer membrane. As the degeneration proceeded, myelin ovoids formed which completely lacked intramembranous particles. The findings suggest that during Wallerian degeneration there is a progression of myelin changes leading to the eventual loss of myelin intramembranous particles. These observations are morphological evidence that Schwann cells remove components from selective portions of their membrane during Wallerian degeneration.     

Abnormal Schwann cell/axon interactions in the Trembler-J mouse

The Trembler-J ( Tr ^J ) mouse has a point mutation in the gene coding for peripheral myelin protein 22 (PMP22). Disturbances in PMP22 are associated with abnormal myelination in a range of inherited peripheral neuropathies both in mice and humans. PMP22 is produced mainly by Schwann cells in the peripheral nervous system where it is localised to compact myelin. The function of PMP22 is unclear but its low abundance (∼5% of total myelin protein) means that it is unlikely to play a structural role. Its inclusion in a recently discovered family of proteins suggests a function in cell proliferation/differentiation and possibly in adhesion. Nerves from Tr ^J and the allelic Trembler ( Tr ) mouse are characterised by abnormally thin myelin for the size of the axon and an increased number of Schwann cells. We report ultrastructural evidence of abnormal Schwann cell-axon interactions. Schwann cell nuclei have been found adjacent to the nodes of Ranvier whereas in normal animals they are located near the centre of the internodes. In some fibres the terminal myelin loops faced outwards into the extracellular space instead of turning inwards and terminating on the axon. In severely affected nerves many axons were only partially surrounded by Schwann cell cytoplasm. All these features suggest a failure of Schwann cell–axon recognition or interaction. In addition to abnormalities related to abnormal myelination there was significant axonal loss in the dorsal roots.     

Fyn tyrosine kinase participates in the compact myelin sheath formation in the central nervous system

The cellular mechanisms for spiral wrapping and compaction of myelin sheaths by oligodendrocytes are not known yet. In this study, we examined the role of fyn tyrosine kinase, which could be responsible for molecular events during the stage of myelination in the CNS. Western blot and immunohistochemical analyses revealed that fyn-deficient mice have significantly lower levels of myelin basic protein (MBP), which is required for intracellular membrane adhesion parts so-called major dense line (MDL) and thought to be essential for the stability of myelin sheath. Electron microscopy verified that the myelin ultrastructure could be used to distinguish fyn-deficient mice from wild-type mice, showing a thin and redundant myelin sheath in the corpus callosum. Further, the electron-dense 'major' line in myelin from the purified myelin fractions remained condensed, and myelin compaction was split opened in fyn-deficient mice. To determine whether there was a change in the microheterogeneity of MBP due to a post-translational event we first investigated peptidylarginine deiminase (PAD), which is an enzyme that converts arginine residues in peptides to citrulline residues. PAD immunoreactivity was observed both in the myelin from fyn-deficient and wild-type mice. By Western blot analysis we found an increase of the citrullined form of MBP. In addition, MBP from fyn-deficient mice did weakly induce vesicle aggregation properties of MBP-mediated adhesion. We concluded that although oligodendrocytes from fyn-deficient mice are able to wrap around the axon, they are unable to form compact myelin due to decreased MBP level and the presence of increased citrullinated MBP.     

Dorsal root ganglion neurons induce transdifferentiation of mesenchymal stem cells along a Schwann cell lineage

It has been reported that mesenchymal stem cells (MSCs) can transdifferentiate into Schwann cell-like cells by a series of treatments with a reducing agent, retinoic acid and a combination of trophic factors in vitro, and can transdifferentiate into myelin-forming cells to repair the demyelinated rat spinal cord in vivo. We now report that when co-cultured with dorsal root ganglion (DRG) neurons, MSCs were induced to transdifferentiate into Schwann cell-like cells that had ensheathed DRG axons. Following differentiation, MSCs underwent morphological changes similar to those of cultured Schwann cells and express GFAP and S100, the marker of Schwann cells. Moreover, 6 weeks later, MSCs wrapped their membrane around DRG axons. Further, initiation of myelination was observed in the co-cultured DRG neurons, which was determined by signals to MBP and this initiation of axon myelination by MSCs is similar to that of Schwann cells. However, electron micrographs show that no compact myelin was present in the MSCs co-cultures, whereas the Schwann cells co-cultures had formed a multilammelar myelin sheath around the axon. These indicate that the release of cytokine by DRG neurons may promote the transdifferentiation of MSCs, but is not sufficient to elicit compact myelination by transdifferentiated MSCs. These results improve our understanding in the mechanism of MSC transdifferentiation, and the mechanism underlying ensheathment and myelination by transdifferentiated MSCs.     

Double myelination of axons in the sympathetic nervous system of the mouse. I. Ultrastructural features and distribution

Summary This study has examined the structural features and distribution of doubly myelinated axons in normal adult and aged mice. Investigation focused on the superior cervical ganglion (SCG) and paravertebral sympathetic ganglia, which were extensively serial-sectioned for light and electron microscopy. In the SCG, the principal features of doubly myelinated regions were that an apparently normal myelinated axon was enclosed for part of its length by an additional (outer) myelinating Schwann cell. The separate nature of the inner and outer Schwann cells was emphasized by the consistent presence of individual nuclei in each, and by the presence of endoneurial space, often containing collagen fibrils, between the inner and outer cells. In some cases more than a single outer Schwann cell was present, arranged serially along the inner myelinated fibre. While double myelination forms through a mechanism involving displacement of an original myelinating Schwann cell by an interposed Schwann cell (see companion paper), we here provide evidence that in some instances the outer Schwann cell fails to retain any direct axonal contact, either with the axon centrally enclosed within the configuration or with any neighbouring axon. In contrast to the rat, delicate cytoplasmic processes often extended from the lateral extremes of outer Schwann cells. However, again no evidence for axonal contact was found, and similar processes also extended from the paranodal region of some singly myelinated non-displaced Schwann cells. Without exception the outer myelin sheath remained structurally intact, and characteristically underwent a series of conformational changes (progressive infolding of the paranodes and new areas of myelin compaction) which infer a continuing capacity of the outer Schwann cell to translocate myelin-specific components in a co-ordinated manner. A basal lamina was always present on the abaxonal plasma membrane of the outer cell, but not on the adaxonal surface except in areas involved in infolding, thus retaining the polarity which existed at the time of displacement from the axon. At single cross-sectional levels through the SCG, up to approximately 4% of myelinated axons were involved in double myelination. Double myelination was not detected in the sciatic nerve or in the paravertebral ganglia, thus indicating a predilection for the SCG as a site of development of these configurations. Though not challenging the role of the axon in initiating the formation of myelin, these data indicate that in this tissue myelin maintenance does not require direct contact between axonal and Schwann cell plasma membranes.     

Ballooning of myelin sheaths in normally aged macaques

In aged animal brains, a variety of holes are formed in the neuropil. One type of hole, here designated as the myelin balloon, is an abnormality of the myelin sheath and is found in a number of diverse sites in the brain. Profiles of myelin balloons display rather smoothly rounded peripheral contours and typically range up to 10 m in diameter, although exceptionally large examples may be twice this size. The balloons are bounded by lamellae of myelin, and to accommodate the contents of the balloon, the myelin sheath becomes split at the intraperiod line. Since the intraperiod line is formed by the apposition of the outer faces of the myelin-forming plasma membrane, the contents of the myelin balloons are, in effect, in continuity with the extracellular space, and it is suggested that the contents of the balloons are fluid, with the fluid exerting an outward pressure on the walls of the balloons to produce their spherical shapes. Myelin balloons are not only produced during aging but also occur in a number of genetic strains of mice and in a number of human disease states. They thus represent a non-specific, though distinctive and common, alteration of the myelin sheath and are a reflection of the fact that under a variety of conditions, including normal aging, oligodendrocytes are unable to maintain the integrity of their sheaths.