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Background and Aim: A murine model mimicking osmotic demyelination syndrome (ODS) revealed with histology in the relay posterolateral (VPL) and ventral posteromedial (VPM) thalamic nuclei adjoined nerve cell bodies in chronic hyponatremia, amongst the damaged 12 h and 48 h after reinstatement of osmolality. This report aims to verify and complement with ultrastructure other neurophysiology, immunohistochemistry, and molecular biochemistry data to assess the connexin-36 protein, as part of those hinted close contacts.This ODS investigation included four groups of mice: Sham (NN; n  = 13), hyponatremic (HN; n  = 11), those sacrificed 12 h after a fast restoration of normal natremia (ODS12h; n  = 6) and mice sacrificed 48 h afterward, or ODS48 h ( n  = 9). Out of these, thalamic zones samples included NN ( n  = 2), HN ( n  = 2), ODS12h ( n  = 3) and ODS48h ( n  = 3)., Results: Ultrastructure illustrated junctions between nerve cell bodies that were immunolabeled with connexin36 (Cx36) with light microscopy and Western blots. These cell's junctions were reminiscent of low resistance junctions characterized in other regions of the CNS with electrophysiology. Contiguous neurons showed neurolemma contacts in intact and damaged tissues according to their location in the ODS zones, at 12 h and 48 h post correction along with other demyelinating alterations. Neurons and ephaptic contact measurements indicated the highest alterations, including nerve cell necrosis in the ODS epicenter and damages decreased toward the outskirts of the demyelinated zone., Conclusion: Ephapses contained C × 36between intact or ODS injured neurons in the thalamus appeared to be resilient beyond the core degraded tissue injuries. These could maintain intercellular ionic and metabolite exchanges between these lesser injured regions and, thus, would partake to some brain plasticity repairs.

Background: To identify capsular components of neuromuscular spindles in man by means of immunohistochemistry., Methods: Investigation of histologically observed neuromuscular spindles in surgical specimens with the use of markers for sheath cells and basement membranes., Results: Epithelial membrane antigen and CD34 immunoreactivities were found in the outer and inner capsular layers, respectively. S-100 protein was not expressed in the capsules and there was more collagen type IV than laminin., Conclusions: Cells resembling perineurial cells and endoneurial fibroblasts, and basement membrane rich in collagen type IV comprise the capsules of neuromuscular spindles.

Myelinated nerve fibers formed by the processes of LV spinal ganglion neurons were studied in two lines of rats selected according to high and low thresholds of nerve system excitability to electric current. Before ultrastructural study the fibres have been treated with potassium pyroantimonate. It was demonstrated that specific dense precipitate was deposited in the nodes of Ranvier of the nerve fibers of rats with low excitability thresholds; this precipitate was not found in the fibers of rats with high thresholds. It is suggested that deposition of precipitate is indicative of a high density of sodium channels in neurilemma, i.e. of high functional activity of nerve fibers.

Objective: The purpose of our study was to evaluate the effectiveness of MR imaging for showing the intrinsic anatomy of a peripheral nerve. Cadaver wrist specimens that included the median nerve were imaged with MR imaging at 3 T, then sectioned, stained, and inspected grossly and microscopically. The size, shape, and signal intensity of the sheath and axonal structures in the median nerve were identified in MR images by comparison with anatomic sections., Conclusion: This study suggests that MR imaging with sufficiently high-resolution techniques shows the internal structure of peripheral nerves. These results suggest that MR imaging may be a means to distinguish neuritis, tumor, degeneration, or fatty proliferation in a peripheral nerve and to evaluate the nerve before microsurgical anastomosis.

Ribosome-lamella complexes are occasionally seen in neoplastic disorders of the hematopoietic system, particularly in hairy cell leukemia. However, these structures are rare in epithelial and mesenchymal cells. In this report three tumors from a patient with neuro-fibromatosis (von Recklinghausen's disease) are examined by electron microscopy. Ribosome-lamella complexes were seen in the perineurial cells. This may be the first report of these complexes in nerve sheath tumors.

The authors conducted a morphometric investigation of the histopathological alterations in myelinated fibers (MFs) of the nerve roots of C-6, which showed macroscopic indentation, presumably due to cervical spondylotic radiculopathy. In six cadavers, designated as the radiculopathy group, in which the nerve roots of C-6 showed indentation due to compression on one side (indented side) and the remaining nerve roots (normal side) showed a normal appearance macroscopically, morphometric findings of the nerve roots and the MFs on both the indented and normal sides were evaluated and subjected to blind comparison. Seven cadavers with normal-appearing C-6 nerve roots served as controls. In the control group, there were no differences in the morphometric parameters: that is, total transverse fascicular area, total number of small and large MFs, and relationship between myelin thickness and the radius of the axon between the right and left sides in either the ventral or dorsal roots. There was no evidence found of axonal degeneration, ongoing demyelination, or loss of MFs in either the ventral or dorsal roots in the radiculopathy group on the indented side. However, there were significantly lower numbers of large MFs per root and significantly thinner myelin sheaths relative to axon size on the indented side compared with those on the normal side in both the ventral and dorsal roots. These findings are characteristic alterations of the MFs produced by chronic low-grade compression.

The histogenesis of cutaneous nerve sheath myxoma (NSM)/cellular neurothekeoma (CNT) is still controversial. In this study, we examined the ultrastructural features of 16 NSM (3 classical, 11 CNT, and 2 mixed NSM/CNT). We classified the cells into 4 groups ultrastructurally. Type I cells were undifferentiated polygonal cells with ovoid nuclei, cytoplasmic microfilaments, and occassionally with microfilament-associated dense bodies. In most cells, the cytoplasmic membrane showed focal membranous densities and occasional basal-lamina-like material. This cell type comprised approximately 90% of CNT. Type II cells were more differentiated, had ovoid or spindled shapes, were rich in intracytoplasmic filaments, and were surrounded by continuous basal lamina. These cells were consistent with Schwann cells and were present in the classical and mixed forms of NSM, and in a single case of CNT. Type III cells had features of perineurial cells and were relatively rare in classical NSM. Type IV cells resembled fibroblasts and were encountered in all variants of NSM. These results support the view that 1) the classical NSM has neural (mainly Schwann cell) differentiation, 2) CNT is predominantly composed of undifferentiated cells with partial features of Schwann cells, smooth muscle cells, myofibroblasts and fibroblasts, suggesting a divergent differentiation, and 3) CNT and NSM represent a histologic spectrum, but in CNT, the neural features are not fully expressed.

Facial paralysis in the newborn may be caused by trauma associated with delivery or by lesions secondary to congenital developmental anomalies. We previously reported the absence of the facial nerve in the horizontal segment of the temporal bone on both sides in a patient with Mobius' syndrome, usually classified as a nuclear developmental defect. In this study we report the findings from 7 additional temporal bones with similar abnormalities. A study of 42 temporal bones from infants with various congenital anomalies revealed nine temporal bones in which the facial nerve disappeared mid-way through the course of the temporal bone. Some of the nerve fibers suggested regeneration after necrotic lesions induced during the perinatal period.

We have characterized T-cadherin, a glycoprotein of 95 kd from chick embryo brain by expression cloning and sequencing of corresponding cDNAs. The nucleotide sequence predicts a novel cadherin cell adhesion molecule in the nervous system. Surprisingly, the isolated cDNAs do not encode the cytoplasmic region conserved in other cadherin subclasses. Biochemical analysis revealed that this truncated (T) cadherin is attached to the neuronal plasma membrane through a glycosyl phosphatidylinositol anchor. T-cadherin is a component of different neuronal populations and is expressed in a temporally and spatially restricted pattern during axon growth. These results are consistent with a putative role of T-cadherin in axon growth and guidance.

A case of dermal nerve sheath myxoma on the left palm of 26-year-old Japanese man is presented. We examined this tumor by light and electron microscopies. On light microscopic examination, this tumor was characterized by a multilobulated myxoid tumor composed of stellate cells in an abundant mucous matrix. This histological feature resembled previously reported cases of dermal nerve sheath myxoma. On electron microscopic examination, the findings of these tumor cells were similar to those of the perineurial cells. These microscopical examinations have provided evidence for an origin from perineurial cells rather than Schwann cells.

Every nerve must have the capacity to adapt to different positions by passive movement relative to the surrounding tissue. This capacity is provided by a gliding apparatus around the nerve trunk. There is another level of gliding provided by the interfascicular epineurium which allows the fascicles to glide against one another. The clinical significance of the gliding apparatus in the context of external and internal neurolysis and nerve repair is discussed. An explanation is offered for the occurrence of the so-called meander-like deformity of fascicles, seen in nerve entrapment syndromes.

There is no firm glial isolation at the border of the intramural ganglion. All elements of the intramural ganglion, situating on its border, among them neurons and dendrites, have contacts with the interstitium and are separated from the latter by a common basal membrane. The insterstitium together with its collagenous fibers can protrude into the ganglionic neuropil. There is no difference in the ultrastructure of superficial and deep ganglionic components. Intercellular canals of the neuropil form a single transporting system with the periganglionic space. Free synaptic terminals of the vegetative neural fibres open into the periganglionic fissure and into the intercellular canals of the neuropil; that conditions an increased contact of neurotransmitters in the tissue fluid, washing neurons of the ganglion, and a constant extrasynaptic trophic and/or modulating effect on the ganglionic neural cells.

"Dystrophic' mice of the 129/ReJ-Dy strain have a genetic defect affecting Schwann cell proliferation. Spinal nerve roots of these animals contain myelinated and unmyelinated axons in addition to groups of large "amyelinated' axons. In affected regions of the spinal roots, myelinated axons are missing their myelin sheaths. Where the myelination terminates or begins, half-nodes are created. Freeze-fracture analysis of these half-nodes shows that only the myelinated side contains rows of dimeric particles in the axonal P-face of the paranode. The P-face on the amyelinated side of a half-node, and the remainder of the amyelinated axon. contains a dense even distribution of particles, many of which are the size of dimeric-particle subunits, but only a few of which are arranged into short rows. As the long circumferential rows are not found on the unmyelinated side of the myelinated side of the half-node we conclude that the paranodal rows of dimeric particles are dependent upon myelination for their organization.

The long nonmyelinated portion of the unipolar process of spinal ganglion cells resembles in many instances the perikaryon and is characterized by the following structural pecularities: 1. The surface membrane displays numerous invaginations and evaginations, which interdigitate with folds of the investing satellite cells, resulting in a considerable increase of the area of intercellular contact. The intercellular gap frequently widens to intercellular cisternae. 2. The axolemma of the most distal part of the nonmyelinated portion is undercoated by dense material and thus resembles the "initial segment" of multipolar nerve cells. 3. The unipolar process of spinal ganglion cells shows a conspicuously high density of neurotubules. The neurotubules frequently collect into fascicles in the same manner as was described for the initial segment of multipolar nerve cells. 4. The nonmyelinated part as well as the first internodes of the myelinated part of the unipolar cell process contain a highly developed axoplasmic reticulum, many-partly huge-mitochondria, a striking number of dense bodies and clusters of ribosomes. The myelin sheath of the first internodes of the unipolar process is unusually thin in relation to their axon diameters. At successive internodes the thickness of the myelin sheath increases stepwise, the Schwann cell loops of the paranodal zones changing their appearence correspondingly. In the great number of ultrathin sections scanned in this study, not one synapse was found, neither in the unmyelinated initial segment nor in the soma.

Instead of the single-channel pore proposed earlier [Wooldridge, D. E. (1984) Proc. Natl. Acad. Sci. USA 81, 5609-5612] for the transit of conductance ions through the neural membrane, a pore with a second channel for "influence ions" of calcium or magnesium is considered in this paper. By entering trapping centers at the closed inner ends of the new channels, the influence ions are postulated to alter the rates of chemical reactions that change the configurational state of the gates guarding the inner ends of the nearby conductance channels. This makes the permeability of the conductance channels strongly dependent on voltage. Using a four-state reaction scheme for both the sodium and potassium pore systems, a computer model of the membrane conductance is constructed. When suitable values are assigned to its parameters, the model closely reproduces the results of the Hodgkin-Huxley voltage-clamp and action potential experiments.

On the basis of histological and ultrastructural study of 57 neurilemmomas 4 main morphological types of tumour are distinguished; with a prevalence of the Antoni A type tissue, with a prevalence of the tissue of the transitory zone, prevalence of the Antoni B type tissue and a fibrosing variant of neurilemmoma. A variety of the morphological variants of neurilemmoma is due to different quantitative combination of morphological sings of degeneration and necrosis of tumour cells with the organization of the aseptic necrosis of tumour cells with the organization of the aseptic necrosis by their replacement with a fibrous tissue. The peculiarity of the peripheral nerve blood supply is a premise for the development of tumour cell degeneration.

The macromolecular organization of membranes isolated from the rabbit optic nerve and tract was analyzed using the freeze-fracture technique. A myelin fraction and two axolemma-enriched fractions were prepared from a preparation of myelinated axons isolated by flotation in a buffered salt-sucrose medium. In the myelinated axon preparation, axolemma and myelin membranes were easily identified. Larger areas of the axon membrane and myelin membrane totally lacked intramembranous particles. The particles remaining on the myelin membrane formed patches of evenly distributed elongated and globular particles. In contrast, the particles remaining on the axolemma were globular in shape and tightly clustered. Particle clustering and particle-free areas were not characteristic of either the axolemma or myelin membrane of whole nerves fixed in situ and processed for freeze-fracture. The isolated myelin membrane fraction contained a large number of vesicles completely lacking intramembranous particles. Of the remaining membrane vesicles, profiles with dispersed elongated and globular particles predominated. A small percentage of vesicles displayed intramembranous particles of the same size, shape and clustering pattern as that seen on the axolemma of the myelinated axon preparation. The two axolemma fractions were enriched in membrane containing tightly clustered globular particles. Particle-free vesicles as well as some myelin membrane vesicles were also seen in the axolemma fractions.

A basic voltage-dependent conformational transition mechanism is proposed. It comprises one relatively fast conversion between two individual states which are comparatively slowly coupled with a third state. Having introduced voltage as an additional parameter of state, standard methods of thermodynamics and rate theory are employed to describe the equilibrium and kinetic behavior of the system. In particular, a quantitative discussion is given regarding the asymmetrical displacement currents generated by switching on and off a voltage pulse. Effects of temperature, pulse duration, and application of a conditioning prepulse are examined. The results provide a comprehensive basis for a quantitative analysis of pertinent experimental work. The so far presented measuring data can indeed by very well described along these lines.

Ten examples from W. J. Schmidt's workshop illustrate the method of ultrastructural research before the advent of the electron microscope. The consistency is shown with which the research worker W. J. Schmidt pursued his goal, how he succeeded in making fundamental discoveries with his polarizing microscope, and interpreted the results of his indirect structural investigations in such a forward-looking and clearsighted manner that many of them were confirmed by direct reproduction in the electron microscope. This makes W. J. Schmidt one of the great pioneers of ultramicroscopic structural research. He was an extremely prolific writer. His list of publications numbers 404 scientific contributions, three classical books among them. The last of these three monographs was written in 1971; its title is "Polarizing Microscopy in Dental Tissues"; it deals with the ultrastructure of teeth, a subject which never ceased to attract his attention during the more than 50 years of his career as a scientist. It was his intention to write such a textbook on the ultrastructure of teeth during his retirement, a task which he accomplished in spite of the infirmities of old age, thanks to his unbroken will to work. Another characteristic feature of W. J. Schmidt was his extraordinary insistence on complete independence; there are only four among his 404 publications which he wrote, at an advanced age, jointly with Helmut Ruska, and the dental monograph he published in callaboration with A. Keil. Everything else was entirely his own effort. His capacity for work was almost inexhaustible. In 1937, for instance, he published 18 scientific papers, among them his richly illustrated protoplasm monograph of 388 pages. His inflexible will to observe everything personally and to interpret and edit his findings alone was part of his special intellectual constitution. In discussions he stubbornly defended his point view and considered suggestions and new ideas only after he had tested them with his own experiments. This had already become evident at the beginning of his career in a discussion with Ambronn of the nature of birefringence of organized substances. But once he had accepted something as proved he propounded it with persistence and enthusiasm.

Guinea pig and rat sciatic nerves were fixed with cacodylate-buffered aldehydes and OsO4, and were stained with ferric ion and ferrocyanide. Cytoplasmic surfaces of the non-myelinated nodal axon membrane of A-fibres display distinct electron-dense aggregates of stain. These aggregates were not observed in association with the paranodal or internodal axolemma. The membranes of C-fibres exhibit no staining under these conditions. Thus, the nodal axolemma of normal myelinated fibres is structurally distinct from both the myelinated internodal membrane, and from the axolemma of C-fibres. The ferric ion-ferrocyanide technique may provide a method for marking axonal membrane with normal nodal properties.

In the preceding paper (Kobayashi, T., S. Tsukita, S. Tsukita, Y. Yamamoto, and G. Matsumoto, 1986, J. Cell Biol., 102:1710-1725), we demonstrated biochemically that the subaxolemmal cytoskeleton of the squid giant axon was highly specialized and mainly composed of tubulin, actin, axolinin, and a 255-kD protein. In this paper, we analyzed morphologically the molecular organization of the subaxolemmal cytoskeleton in situ. For thin section electron microscopy, the subaxolemmal cytoskeleton was chemically fixed by the intraaxonal perfusion of the fixative containing tannic acid. With this fixation method, the ultrastructural integrity was well preserved. For freeze-etch replica electron microscopy, the intraaxonally perfused axon was opened and rapidly frozen by touching its inner surface against a cooled copper block (4 degrees K), thus permitting the direct stereoscopic observation of the cytoplasmic surface of the axolemma. Using these techniques, it became clear that the major constituents of the subaxolemmal cytoskeleton were microfilaments and microtubules. The microfilaments were observed to be associated with the axolemma through a specialized meshwork of thin strands, forming spot-like clusters just beneath the axolemma. These filaments were decorated with heavy meromyosin showing a characteristic arrowhead appearance. The microtubules were seen to run parallel to the axolemma and embedded in the fine three-dimensional meshwork of thin strands. In vitro observations of the aggregates of axolinin and immunoelectron microscopic analysis showed that this fine meshwork around microtubules mainly consisted of axolinin. Some microtubules grazed along the axolemma and associated laterally with it through slender strands. Therefore, we were led to conclude that the axolemma of the squid giant axon was specialized into two domains (microtubule- and microfilament-associated domains) by its underlying cytoskeletons.

Under anoxia, ultrastructural alterations were revealed in the cytoplasm of sympathetic axons and their synapses, and in lemmocytes in unmielinated bundles of the frog urinary bladder. Anoxia acts as a stimulator. Six minutes anoxia causes the amplification of activity of nerve bundle organoids: augmentation of the granular ER, the number of ribosomes and pinocytotic vesicles, and swelling of mitochondria in trunk and preterminal areas of nerve bundles in lemmocytes. In par distance synapses, small synaptic vesicles are modified, vacuoles are formed, and mitochondria get swollen; the cytoplasm of lemmocyte processes around the synaptic boutons is swollen, its fibrillar structures and part of ribosomes disappear. Under a prolonged anoxia, the number of ribosomes in nerve bundles is reduced, the mitochondria are getting more numerous but with lower electron density; the big dense cored vesicles are accumulated in synapses. After the frog death due to anoxia, deformation of part of the organoids is seen to accompany these changes.

There is an extensive bundle of dendrites with a rostro-caudal axis in the ventro-lateral lamina of the sixth lumbar segment in each side of the rat spinal cord. Such a bundle has a diameter of about 250 mum and contains over 1400 parallel dendrites, each with a diameter of less than 8 mum, interspersed between neuronal somata. The volume fraction of dendrites in the bundle neuropil is about 55%, the remainder being equally distributed between astrocytes, synaptic boutons, and axons, most of which are unmyelinated. An analysis of the median percentage covering of dendrites by contiguous elements of the neuropil reveals that as the dendrite diameter decreases from 4 to 0.2 mum (mean equals 2 mum), astrocytes increase from 43 to 75%, axons decrease from 21% to zero, boutons decrease from 28% to zero, and dendrites decrease from 10% to zero. There is a mean of 18 synaptic boutons per 100 mum-2 of the overall dendritic surface, but larger boutons tend to be more frequent on larger dendritic profiles. Apposed dendrites and their somata may have either puncta adhaerentia or confronting subsurface cisternae. Synaptic types in the rat are similar to those reported for the cat. The morphological findings are discussed with respect to previously proposed interaction between neural elements.

Ultrastructural changes of the terminal plates of the bushy receptors in the frog urinary bladder have been studied after two hours' exposition in 0.05% novocaine solution and one hour's exposition in 0.05% dicaine and trimecaine solution. During these periods a steady block of the receptor impulse activity develops. The local anesthetics essentially change ultramicroscopic structure of the terminals. The reaction to the anesthetics investigated has both some features in common and certain peculiarities. At each effect three types of changes can be determined, characterized with various degree of rearrangement in neurilemma, neuroplasm and organelles. Each type of the changes is supposed to reflect a certain phase of the plate reactive response. Specificities of the reaction to novocaine are minimal changes of mitochondria, accumulation of glycogen granules, deformity and decreasing amount of vesicles. Under dicaine effect mitochondria do not change, amount of vesicles increases, their form does not change; under trimecaine effect mitochondria undergo most noticeable alterations. The changes of the terminal plates observed are interpreted as adaptive. The effect of the local anesthetics on the receptors is not limited with the blockade of the sodium canals of the afferent fibers, in parallel, biochemical processes, occurring in cytosol of the terminals also change; their morphological manifestations are the ultrastructural changes observed.

The freeze-fractured membrane of the central myelin sheath has three classes of particulate components: (i) Particles inherent to the compact myelin lamellae. These are distributed at random and cleave predominantly with the P (protoplasmic) face. (ii) Particles which comprise the intramyelinic tight junctions. These are arranged in strands and are located at the inner and outer mesaxon, the paranodal loops, the cytoplasmic incisures, and occasionally within the compact regions of the myelin sheath. (iii) Particles localized exclusively at the portion of the paranodal loop membrane involved in the septate-like junction with the axolemma. These are regularly spaced and are organized in parallel rows. In the central myelin sheaths of bullfrogs fixed by perfusion with aldehydes and cryoprotected in 30% glycerol, the randomly distributed particles differ in size and shape from those of the axolemma. They possess a reasonably well defined bimodal distribution with respect to particle shape--most can be described either as globules or as ellipsoids. The globular particles range in diameter from 60 to 150 A. The ellipsoidal particles are 100-200 A long and 15-50 A wide. The total number of particles per square micron on the P face is approximately 1500. About half of these are of the globular type and half of the ellipsoidal type. In poorly fixed specimens, loss of interlamellar adhesion and loss of randomly distributed particles seem to coincide. Evidence is presented against the hypothesis that the tight junctions between compact myelin lamellae represent the radial component of the myelin. The possible relation between the types of particulate components seen in freeze-fracture and the classes of protein isolated from central myelin fractions is briefly discussed.

An investigation was made of ventral and dorsal spinal roots (L7 and S1) in the postnatal life of the rabbit. The development of the Schwann sheath of the nerve fibers was studied and correlated with the distribution and morphological characteristics of the desmosome-like structures connected with them. The association of these structures with constituents of the endoplasmic granular and smooth-surfaced reticulum of the Schwann cell is pointed out. The possibility is discussed of a direct relationship between the inner mesaxons, as well as along the length of the peripheral myelin lamellae. In addition, it is suggested that in some cases already existing membrane formations of the adjacent cytoplasm become attached to the developing Schwann sheaths, which results in branching mesaxons or "poli-mesaxonal" nerve fibers.

It is surprising to ascertain, as did DE LEMNOS and PICK, that the fine structure of rat ganglia has not raised up more interest, If one considers neurons only, S.I.F. and satellite cells are not generally described. Relatively little information is available on the nucleus organization, except nuclear mambran folds (FORSSMANN). Cajal's intranuclear rod shaped inclusions have been omitted or imperfectly studied except by SEITE. However, the presence of the structures testifies the biochemical activity in neurons. In the present ultrastructural investigation, perticular attention was given to the perikaryon fine structure in neurons. Changes in neuronal appearance were estimated in cultures and grafts with reference to morphological features of the perikaryon.

Peripheral nerve transplants are used for the surgical repair of nerve loss. If normally oriented grafts contain branches, regenerating axons may be lost into them, limiting re-innervation and restoration of function in the denervated area. Since axons entering a reversed graft may not be lost in this way, we have investigated whether reverse implantation could enhance the number of axons that reach the distal stump. In young adult rats, a section of sciatic nerve, including one major branch, was removed and reinserted with either the normal or reversed proximo-distal orientation, using microsurgical techniques. After signs of recovery of function, the operated and contralateral unoperated nerves of each animal were fixed and processed for electron microscopy. Regenerated axons were seen to enter branches in normally orientated grafts, and there was substantial loss in the cross-sectional area of the graft distal to the branch termination. Reverse grafts are as well reinnervated by regenerating axons as normally oriented grafts and show a smaller loss in the cross-sectional area, even though the branches of these implants seemed to disappear.

Asymmetrical displacement currents are measured in the absence and in the presence of the lipophilic anion dipicrylamine (DPA) in the extracellular solution of nerve fibers of the frog Rana esculenta. DPA (30 nM--3 microM) enhances the current by a component that has the properties expected for a translocation current of DPA ion across the lipid membrane. Analysis in terms of a single-barrier model yields the translocation rate constant (k), the total surface density of DPA absorbed to the membrane (Nt), and the equidistribution voltage (psi). The value of kappa of about 10(4) s-1 is similar to that for a solvent-free artificial bilayer formed by the Montal-Mueller method. The surface density Nt varies with the DPA concentration as it does in the artificial bilayer, but is about tenfold smaller at all concentrations. The DPA ions sense an intrinsic electric field that is offset by a transmembrane voltage between 0 and 30 mV (inside positive). The part of the axolemma probed by the DPA ion appears as a thin ( less than 2.5 nm), fluid bilayer of lipids. DPA ions seem, however, to be excluded from the major part of the axolemma as if this area is occupied by integral proteins or negative charges.

Benign tumors from sheaths of peripheral nerves (25 neurolemmomas and 6 neurofibromas) were studied. The ultrastructural data presented attest to the principal differences between these tumors and the possibility of their differential diagnosis. The typical signs of tumor cells of neurilemmoma include numerous complicatedly interwoven processes not infrequently forming " pseudomesaxonal " structures, and the presence of permanent basal membrane. These signs indicate the histogenetic relationship of neurilemmoma with cells of the Schann sheath of the peripheral nerve ( lemmocytes ). Neurofibroma tumor cells were characterized by the presence of bipolar thin, long non-interwoven cytoplasmic processes with inconstant basal membrane, desmosome-like contacts between the processes, and pinocytic vesicles in the cytoplasm. The revealed ultrastructural signs indicate the histogenetic relationship of neurofibroma with cells of the perineurium of peripheral nerves.

Analysis of Shiverer central nervous tissue by the freeze-fracture method shows that axoglial junctions of the type found normally in the paranodal region occur commonly despite the gross reduction in myelin. On a substructural level these junctions appear identical to those that form between paranodal oligodendroglial processes and axolemma. On a grosser level, however, they are bizarre in shape, arrangement and distribution. Isolated glial processes, or small sheaves of them, course among axons and form such junctions in an irregular patchy manner, usually without apparent relationship to paranodal regions. These aberrant junctions may be oriented transversely, obliquely or longitudinally with respect to the axonal axis. Axolemmal E face particle accumulations, which characterize normal nodes of Ranvier, are usually not found in the membrane adjacent to the aberrant junctional patches. Thus, axoglial junctional specializations of the paranodal type can form in this mutant in the absence of the myelin proteins that are deficient in Shiverer, and such junctions may appear in areas not related to other paranodal or nodal structures. The relevance of these findings to differentiation of the axolemma and to the neurological defects in this mutation is discussed.

Patches of bushy receptors in the frog (Rana temporaria) urinary bladder cervix were revealed electron microscopically. The patches have no neurilemma, have specific topography and distribution, they vary in their form and size, their ultrastructural features are presented as specialization of cytolemma, mitochondria of a condenced type, a peculiar receptory matrix, various structure of the basal membrane in different areas. Topographic, morphological and ultrastructural data evidence in favour of a sensitive nature of the bushy receptory terminals described.

The active zone is a unique specialization of the presynaptic membrane and is believed to be the site of transmitter release. The formation of the active zone and the relationship of this process to transmitter release were studied at reinnervated neuromuscular junctions in the frog. At different times after a nerve crush, the cutaneous pectoris muscles were examined with intracellular recording recording and freeze-fracture electron microscopy. The P face of a normal active zone typically consists of two double rows of particles lined up in a continuous segment located opposite a junctional fold. In the initial stage of reinnervation, clusters of large intramembrane particles surrounding membrane elevations appeared on the P face of nerve terminals. Like normal active zones, these clusters were aligned with junctional folds. Vesicle openings, which indicate transmitter release, were seen at these primitive active zones, even though intramembrane particles were not yet organized into the normal pattern of two double rows. The length of active zones at this stage was only approximately 15% of normal. During the secondary stage, every junction was reinnervated and most active zones had begun to organize into the normal pattern with normal orientation. Unlike normal, there were often two or more discontinuous short segments of active zone aligned with the same junctional fold. The total length of active zone per junctional fold increased to one-third of normal, mainly because of the greater number of segments. In the third stage, the number of active zone segments per junctional fold showed almost no change when compared with the secondary stage. However, individual segments elongated and increased the total length of all active zone segments per junctional fold to about two-thirds of the normal length. The dynamic process culminated in the final stage, during which elongating active zones appeared to join together and the number of active zone segments per junctional fold decreased to normal. Thus, in most regions, regeneration of the active zones was complete. These results suggest that the normal organization of two double rows is not necessary for the active zone to be functional. Furthermore, localization of regenerating active zones is related to junctional folds and/or their associated structures.

While studying the ultrastructure of rat celiac nodes, it was stated that lemmocytes from the intercellular plexus develop around separate neuronal processes spiral membranes and multilayer membranes in the shape of concentric "musses". Similar membranes but of simpler structure develop perisomatic glyocytes. In old rats these types of glial membranes occur considerably rare. Both varieties of complicated types of glial membranes serve, evidently, for the isolation of preterminal sections of neuronal processes from probable unwanted contacts with other terminals.

Using the squid giant axon, we analyzed biochemically the molecular organization of the axonal cytoskeleton underlying the axolemma (subaxolemmal cytoskeleton). The preparation enriched in the subaxolemmal cytoskeleton was obtained by squeezing out the central part of the axoplasm using a roller. The electrophoretic banding pattern of the subaxolemmal cytoskeleton was characterized by large amounts of two high-molecular-weight (HMW) proteins (260 and 255 kD). The alpha, beta-tubulin, actin, and some other proteins were also its major constituents. The 260-kD protein is known to play an important role in maintaining the excitability of the axolemma (Matsumoto, G., M. Ichikawa, A. Tasaki, H. Murofushi, and H. Sakai, 1983, J. Membr. Biol., 77:77-91) and was recently designated "axolinin" (Sakai, H., G. Matsumoto, and H. Murofushi, 1985, Adv. Biophys., 19:43-89). We purified axolinin and the 255-kD protein in their native forms and further characterized their biochemical properties. The purified axolinin was soluble in 0.6 M NaCl solution but insoluble in 0.1 M NaCl solution. It co-sedimented with microtubules but not with actin filaments. In low-angle rotary-shadowing electron microscopy, the axolinin molecule in 0.6 M NaCl solution looked like a straight rod approximately 105 nm in length with a globular head at one end. On the other hand, the purified 255-kD protein was soluble in both 0.1 and 0.6 M NaCl solution and co-sedimented with actin filaments but not with microtubules. The 255-kD protein molecule appeared as a characteristic horseshoe-shaped structure approximately 35 nm in diameter. Furthermore, the 255-kD protein showed no cross-reactivity to the anti-axolinin antibody. Taken together, these characteristics lead us to conclude that the subaxolemmal cytoskeleton in the squid giant axon is highly specialized, and is mainly composed of microtubules and a microtubule-associated HMW protein (axolinin), and actin filaments and an actin filament-associated HMW protein (255-kD protein).

Ferric ion and ferrocyanide were used to stain central nervous tissue from the spinal cords of rats following fixation in cacodylate-buffered aldehydes. At the nodes of Ranvier in myelinated fibers, the stain was localized primarily on the inner surface of the unmyelinated nodal axolemma, as had been reported previously for peripheral nodes. Unmyelinated initial segments of myelinated neurons were similarly stained, but the axon hillock, cell body and dendrites were not stained. Synapses also exhibited no staining. Details of stain localization and comparison of these results with other ultrastructural data suggest that the stain is specific for the node of Ranvier and the axon initial segment, and are consistent with the idea that the axolemma at these sites may be structurally different from the cell membrane in other regions of the neuron, including paranodal and internodal regions of the axon. The electron-dense substance underlying the cytoplasmic surface of the membrane at the nodes and initial segments may represent a substrate that serves to bind together membrane structures in specialized regions of the axolemma.

The size and branching structure of the dendritic tree were studied in nine type-identified triceps surae alpha-motoneurons that were labeled intracellularly with horseradish peroxidase and reconstructed from serial sections in the light microscope. The average total membrane area (AN) for motoneurons of type S (slow-twitch) motor units was about 22% smaller than AN for cells of type F units (including both FF and FR motor unit types in this category) (480.1 X 10(3) microns 2 vs. 617.7 X 10(3) microns 2, respectively). Systematic correlations were found between stem dendrite diameter and three measures of dendritic size: dendrite membrane area, combined dendritic length, and number of terminations. All of these correlations were significantly different for the dendrites of F and S motoneurons. Power-function relations between stem diameter and dendritic membrane area were used to estimate AN for a sample of 79 type-identified motoneurons. Mean estimated AN values were significantly different for the F and S motoneuron groups, despite a large overlap in AN values between these groups. The branching structure of dendrites of F and S motoneurons also showed clear differences. Type S motoneuron dendrites showed less-profuse branching and a more-even radial distribution of branch points than found in type F cells. Examination of two forms of the "3/2 power rule" for the relation between the diameters of parent and daughter dendritic branches at branch points showed that the dendrites of type S motoneurons conform less well with the anatomical constraints necessary to represent binary branching trees as equivalent cylinders than do dendrites of type F cells. There was no systematic difference between F and S motoneuron dendrites in the degree of asymmetry of first-order daughter trees. The results overall indicate that the dendrites of F and S motoneuron groups are structurally different, giving rise to a systematic difference in AN between these groups. Such structural differences suggest that the F and S groups of alpha-motoneurons can be viewed as intrinsically distinct cell types and not just large vs. small variants of the same cell species.

The central cells of substantia gelatinosa are small spindle-shaped neurons with an average perimeter of 26.8 micrometers and an average cross-sectional area of 44.9 micrometers2. No difference has been found between neurons of C1 and those of the lower cervical segments. It was calculated that axo-somatic synapses are about 20 per cell and generally occur in small clusters of 3-4 synapses each. Despite their low number they are probably of functional relevance when compared to the small size of the somata of substantia gelatinosa neurons. Most of the synaptic boutons contain pleomorphic electronlucent vesicles and have symmetrical synaptic junctions. Large granular synaptic vesicles also occur intermingled with the electronlucent ones. Sites of somato-dendritic and somato-somatic membrane apposition are fairly extensive: they over about 8% of the cellular perimeter. The possibility of cellular interactions or material interchanges at these sites of direct membrane apposition is suggested. Somatodendritic synapses were occasionally observed. Although the neuropil is indeed the main target of the different inputs to the substantia gelatinosa and the site of most of the local circuits, the somata may too be involved in synaptic transmission and possibly in intercellular coupling.

Synaptic relationships between neuronal processes have been studied in thick sections (up to 1.0 mum) with high voltage electron microscopy. Some of the problems that arise in the study of such thick sections have been considered. (1) Where two parts of a single process are in continuity through a narrow stalk, as a dendritic stem and one of its spines, the continuity can generally not be seen if the stalk is included within the section or is crossed in the section by other processes. (2) Membranes that are viewed face-on cannot be seen in the thick sections and, therefore, processes belonging to two separate nerve cells may appear as though they are in cytoplasmic continuity. (3) Synaptic junctions that are viewed face-on cannot be seen in the thick sections, and junctions lying close to the same angle are also invisible; the relative number of invisible junctions depends upon a number of factors, including the section thickness and the size and contrast properties of the junctional specializations. Two methods of marking neuronal processes have been explored: (1) degenerating axons that are undergoing the dark degenerative change can be recognized readily within the thick sections. However, they stand out less clearly than in thin sections. (2) Neuronal processes that have been impregnated by an extra-rapid modification of the Golgi method can be easily traced in thick sections. It is possible to study some of the synaptic contacts made by these impregnated processes, especially if the processes have been only lightly impregnated or if the silver dichromate deposits are secondarily removed.