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Biomedical subjects

J Rosenbluth

Publications and source records attributed to J Rosenbluth.

At least 55 records · Page 3Linked to original sources

Structural specializations in cat of chronically demyelinated spinal cord axons as seen in freeze-fracture replicas.

Axons in chronically demyelinated spinal cord lesions, induced by ethidium bromide injection, display patches of intramembranous particles and indentations resembling nodal and paranodal axolemmal specializations respectively. Both occur in the marginal region of the lesions where the demyelinated axons are intimately associated with astrocytic and oligodendrocytic processes, and probably correspond to the aberrant node-like and paranodal junctional complexes seen in thin sections of this region. Demyelinated axons in the center of the lesion, which are not in contact with glial processes, do not display these membrane specializations.

Animals↗

Outbreak of JK diphtheroid infections associated with environmental contamination.

The group JK diphtheroid organism is a multiply resistant opportunistic pathogen which infects immunocompromised patients sporadically. We describe the first reported outbreak of JK diphtheroid infections, in which four cases of bacteremia and one Hickman catheter site infection occurred during 4 weeks on a hematology ward. On this ward, JK diphtheroid was recovered from 17 of 39 patients, 10 of 17 30-ft3 (0.840-m3) air samples, surfaces in 9 of 13 patient rooms, and hands of 4 of 22 personnel. Previously identified risk factors for JK diphtheroid sepsis (male gender, broad-spectrum antibiotic therapy, granulocytopenia, and prolonged hospital stay) were present in infected patients but did not distinguish them from patients who were only colonized. Emphasis on aseptic practices was associated with termination of the outbreak and negative hand cultures from personnel, despite continued patient colonization and environmental contamination.

Air Microbiology↗

Axolemmal differentiation in myelinated fibers of rat peripheral nerves.

In developing rat peripheral fibers, nodal specialization appears early, prior to myelin compaction, and is first detected as a junction between the axon and the overhanging Schwann cell process characterized by a uniformly wide (approximately 18 nm) intercellular gap containing a patchy dense substance and a cytoplasmic undercoating subjacent to the axolemma. The gap width is rather consistent but the axolemmal undercoating is more variable and lower in density than that found at more mature nodes of Ranvier, and it is also highly variable in length, ranging from 0.5 to 3 micron. The outermost Schwann cell layer is usually prominent with a large volume of cytoplasm and many organelles. In freeze-fracture replicas, modal specializations are characterized by accumulations of large (approximately 10 nm) particles in the axolemma, especially the E face, but immature nodes generally have a lower particle concentration than mature nodes. No node-like particle aggregates have been found in axons not intimately associated with Schwann cells. Mature paranodal axon-Schwann cell junctions are usually formed first by the loops closest to the node and are characterized by a 2-3 nm gap between the apposed membranes, periodic intercellular densities (transverse bands) in the gap and cisternae flattened against the junctional Schwann cell membrane. The loops further removed from the node display a wider gap containing irregularly spaced or diffuse intercellular densities, or none. Mature junctions appear relatively late in the rat, and it is not unusual to find developing nodes with several Schwann cell loops present that do not indent the axolemma significantly and are not associated with the paracrystalline pattern characteristic of the mature junctional axolemma. In such instances, the nodal particle aggregates do not have sharply circumscribed boundaries. The majority of the developing nodes are asymmetric with one paranodal segment more mature than the other.

Animals↗

Anionic sites on the surface of frog ependymal astrocytes and mouse ependymal cells.

The binding of colloidal iron hydroxide (CI) and ruthenium red (RR) to the plasma membrane of frog ependymal astrocytes was examined by electron microscopy. Positively charged CI and RR bind to the external surface of the plasma membrane of all parts of the ependymal astrocyte. Prior treatment with neuraminidase markedly reduces the number of bound CI particles, suggesting that the sialic acid of carbohydrates associated with the cell surface is responsible for much of the CI binding. Comparable observations were made on mouse ependymal cells. These findings indicate that an anionic, carbohydrate-rich cell coat occurs on the plasma membrane of amphibian ependymal astrocytes and mammalian ependymal cells. This cell coat may be related to transport, barrier, or receptive functions of ependymal cells.

Animals↗

Development of nodal and paranodal membrane specializations in amphibian peripheral nerves.

Peripheral nerves from the hind legs of frog tadpoles were examined in order to ascertain the pattern of development of nodal and paranodal specializations in myelinated fibers. In thin sections the earliest detectable node-related specializations resemble "intermediate" junctions between axons and Schwann cell processes. These occur in individually ensheathed axons near the edges of the sheath segments and could represent early nodal or paranodal components or transient structures. The characteristic nodal "undercoating" is indistinct and highly variable in thickness in immature fibers and its density is lower in developing nodes than in adult nodes. Corresponding freeze-fracture replicas of developing axons demonstrate aggregates of nodal E face particles whose concentration is lower than that in the adult. Such aggregates usually occur immediately adjacent to Schwann cell indentations, even though early in development the latter may not exhibit the paracrystalline pattern seen in the adult paranodal axolemma. On rare occasions, node-like particle aggregates and presumptive nodal undercoatings have been observed without recognizable paranodal junctions or indentations nearby. However, neither specialization has been found in axons not individually ensheathed by Schwann cells. Paranodal Schwann cell loops are widely separated and irregularly arranged in the developing nodes, and the paranodal regions flanking a node usually mature asymmetrically. Differentiated paranodal junctions appear early in axons ensheathed by only a few loose Schwann cell lamellae. However, such junctions are not formed by all paranodal loops; they consistently appear first in the loops close to the node and only later in those further removed. No junctional specialization has been observed in either the axolemma or the Schwann cell membrane without the close association of the other.

Aging↗

Axoglial junctions in the mouse mutant Shiverer.

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.

Animals↗

Ependymal astrocytes in the frog cerebellum.

We have examined the ependymal astrocytes of the frog cerebellar cortex in thin sections and freeze-fracture replicas. The somata border the fourth ventricle and give rise to basal processes whose daughter branches cross the molecular layer and terminate as subpial endfeet. Irregular lamellar appendages arise from the basal processes and their branches. In the molecular layer the appendages selectively ensheath apposed parallel fiber boutons and Purkinje cell dendritic spines. Other appendages ramify throughout the neuropil, some contributing to extensive pericapillary sheaths. Freeze-fractured ependymal astrocyte plasma membrane consistently has a greater concentration of intramembranous particles (IMPs) and IMPs of larger mean size than neuronal plasma membrane in the same replicas. Like the astrocytes of the mammalian central nervous system, frog ependymal astrocytes form numerous gap junctions with each other. However, orthogonal arrays of IMPs ("assemblies") were not observed. Ependymal cells in the frog cerebellum combine the morphology, and probably the functions, of both ependymal cells and astrocytes.

Animals↗

Freeze-fracture approaches to ionophore localization in normal and myelin-deficient nerves.

(1) The principal result of freeze-fracture studies of myelinated axons is that the axolemma is clearly not uniform in its structure, but rather is highly differentiated in both paranodal and nodal regions. Thus, it is no longer correct to assume that the special physiological properties of myelinated nerve fibers derive only from the presence of the myelin sheath. The inhomogeneity of the axolemma must also be taken into account. (2) The nodal axolemma is characterized by a population of large intramembranous particles primarily in the E fracture face that may correspond to the voltage sensitive sodium channels known to be concentrated there. (3) Significant numbers of such particles also frequently occur in paranodal "lakes" and in the internodal axolemma immediately adjacent to the paranodal region. These are probably accessible, albeit slowly, by way of the narrow extracellular cleft between the paranodal junctional membranes. (4) In the absence of ensheathment by myelinating cells, axons fail to develop normal nodal and paranodal membrane specializations. (5) When ensheathed by abnormal myelinating cells, corresponding abnormalities develop in both nodal and paranodal specializations of the axolemma. (6) Demyelination results in dedifferentiation of axolemmal specializations. (7) It is concluded that development and maintenance of normal axolemmal differentiation requires interaction of the axon with myelinating cells. These cells thus serve not only to produce myelin but also to regulate axolemmal differentiation. Alterations in axolemmal structure following demyelination may significantly affect the physiological properties of the axons. Specifically, ionophore redistribution may underlie the development of either continuous or nonuniform conduction in some demyelinated fibers.

Animals↗

Central myelin in the mouse mutant shiverer.

The spinal cord, optic nerves, and cerebellum of the mouse mutant Shiverer were examined by electron microscopy of thin sections. Although central nervous system myelin is grossly deficient in amount, none of its basic structural elements are missing. Regions of compact myelin can be found composed of several layers of alternating major dense lines and intermediate lines repeating with normal periodicity. The "radial component" consisting of periodic thickenings of the intermediate line aligned through several lamellae was also identified. Axoglial junctions characteristic of the type found in paranodal regions are present in greater than normal numbers but occur in aberrant locations. Myelin sheaths have marked reduced numbers of lamellae, which often contain cytoplasm, terminate in cytoplasmic "loops" within and around myelin sheaths, and do not completely encircle axons. In addition, membranous debris appears within neuronal and glial profiles, suggesting some degree of myelin breakdown. Thus, the protein lacks in this mutant appear not to be associated with discrete deficiencies of specific structural components but rather with a variety of quantitative changes and irregularity of form.

Animals↗

Nodal and paranodal membrane structure in complementary freeze-fracture replicas of amphibian peripheral nerves.

Complementary freeze-fracture replicas of frog peripheral nerves have revealed new details of membrane structures at the node of Ranvier and paranodal axon-Schwann cell junction. At the node both E and P fracture faces of the axolemma have high particle concentrations (approximately 1350/sq. micron and 1600/sq. micron respectively) and these particles do not overlap when tracings from the respective fracture faces are superimposed. A high proportion of the E face particles are large (> 9.5 nm) and cast long shadows while the proportion of large particles in the P face is much lower. In the paranodal region the diagonal pattern of parallel rows in the junctional axolemma always has the same orientation within a given fracture face. In the E face, the parallel rows form a positive (+ 30 degrees) angle to the groove below and in the P face, a negative (-30 degrees) angle to the ridge above. This implies that the diagonal pattern derives from asymmetric subunits that are able to associate along only one axis and are unable to 'flip over' with respect to the junctional membranes.

Animals↗

Freeze-fracture study of the postsynaptic membrane of the cerebellar mossy fiber synapse in the frog.

We have examined the postsynaptic membrane of the synaptic junctions of frog cerebellar mossy fibers by electron microscopy of freeze-fracture replicas and thin sections. The intramembranous particles (imps) in the E fracture face of the postsynaptic membrane are approximately 10 nm in size and form conspicuous aggregates which we classified as macular, annular, or anastomotic in form, according to the occurrence and placement of imp-free "windows" within the aggregate. The size and shape of the aggregates appear related in that the area of macular aggregates is consistently smaller than the area of annular or anastomotic aggregates. Measurements of aggregate area range from 0.06 to 0.75 micrometer2. The variable size and shape of the imp aggregate in the postsynaptic membrane sets it apart from other excitatory synapses in the central nervous system, where macular aggregates are usually described. Examination of serial thin sections suggests that the shape of the postsynaptic density is equivalent to that of the imp aggregate observed in the postsynaptic membrane by freeze-fracture. This supports the notion that the region of postsynaptic membrane associated with the postsynaptic density in thin sections corresponds to the particle-rich regions of E face membrane observed in freeze-fracture replicas.

Animals↗

Peripheral myelin in the mouse mutant Shiverer.

The mouse mutant Shiverer has been shown previously to lack myelin basic protein and other myelin proteins in both the peripheral and central nervous systems. Examination by electron microscopy shows that the peripheral nervous system, in contrast to the markedly abnormal central nervous system, is grossly normal. Myelin sheaths are of the usual thickness and exhibit normal periodic structure consisting of alternating major dense and intermediate lines. Subtle abnormalities do occur, however, consisting of increased numbers of cytoplasm-containing lamellae, aberrant terminations of myelin lamellae in internodal regions, invagination of the axon by the inner tongue of the myelin sheath, myelin debris in both axon and Schwann cells, and disruption of outer myelin lamellae. Such changes have been seen previously in various types of neuropathy and are not pathognomonic of the Shiverer mutation. Despite the absence of myelin basic protein, the peripheral manifestations of this gene are relatively minor and probably not severe enough to compromise peripheral nerve function significantly.

Animals↗

Aberrant axon-Schwann cell junctions in dystrophic mouse nerves.

'Amyelinated' axons in the spinal roots of dystrophic mouse nerves lack typical nodal and paranodal membrane specializations. However, at the periphery of the amyelinated bundles some of the naked axons form aberrant junctions with Schwann cells belonging to neighbouring myelinated axons. These junctions are characterized by a narrow intercellular cleft containing regularly-spaced densities that closely resemble the 'transverse bands' found at paranodal axoglial junctions with respect to both configuration and spacing. In addition, the Schwann cells sometimes extend fingerlike projections towards amyelinated axons in regions where the axolemma has a dense cytoplasmic undercoating. Such regions resemble nodes of Ranvier, where Schwann cell processes interlace over the axolemma. Freeze-fracture replicas show no typical nodal or paranodal membrane specializations in the amyelinated fibres where they are apposed to each other. However, isolated paracrystalline patches of membrane occur corresponding to the aberrant junctions between amyelinated axons and Schwann cells at the periphery of the bundles. The observations show that structural differentiation of the axolemma occurs only where axons are in intimate contact with myelinating cells and does not develop independently in the amyelinated regions. Sodium channels, which are normally concentrated in the specialized nodal membrane, are, therefore, probably distributed uniformly along the amyelinated axon segments that show no sign of such regional differentiation. In addition, it is shown that Schwann cells are capable of forming specialized junctions with more than one axon at the same time.

Animals↗

Glial membrane specializations in extraparanodal regions.

Previous freeze-fracture studies of central myelinated nerve fibres have demonstrated a distinctive junction in the paranodal region formed between the terminal loops of the glial cell and the axolemma. This unique junction is characterized by the presence of diagonally oriented rows of particles in the P face and to a lesser extent in the E face of the glial cell and an equivalent pattern in the axolemma. In both, the rows are spaced at 250--300 A intervals. Although this junction was originally thought to be peculiar to the paranodal region, examples of the same pattern have now been seen in extraparanodal regions in the central nervous system where they appear as circumscribed patches of membrane exhibiting a pattern identical to that in the paranodal glial loops. All examples found were in the immediate vicinity of myelinated nerve fibres and in one case the membrane containing the specialized patch was identified as a lamella of a myelin sheath. These observations constitute evidence that this distinctive membrane specialization is not limited to the paranodal axoglial junction but can also be found in glial membrane specialization is not limited to the paranodal axoglial junction but can also be found in glial membranes not in immediate contact with the specialized membrane of the paranodal axolemma.

Animals↗

Particle arrays in earthworm postjunctional membranes.

Analysis of freeze-fractured earthworm body wall muscle reveals distinctive trough-shaped concavities in the protoplasmic leaflet of the muscle cell membrane which contain diagonally oriented rows of particles sometimes in highly ordered arrays. The troughs correspond to the concave postjunctional patches of sarcolemma seen previously in thin sections of myoneural junctions identified as cholinergic, and the intramembranous particles within the troughs correspond in concentration and arrangement to granular elements present in the outer dense lamina of the postjunctional membrane which were interpreted as acetylcholine receptors. The freeze-fracture data provide a more accurate picture of the arrangement of these putative receptors within the plane of the membrane, and indicate also that they extend into the membrane at least as far as its hydrophobic layer.

Animals↗

Intramembranous particle distribution at the node of Ranvier and adjacent axolemma in myelinated axons of the frog brain.

The plasma membrane of myelinated axons in the frog brain has been examined by the freeze-fracture technique. The cytoplasmic leaflet of the axolemma contains numerous randomly distributed particles in nodal and internodal regions but relatively fewer particles in the axoglial junctional portion of the paranodal region. Particle distribution is even less uniform in the outer leaflet of the axolemma, which contains a low concentration of particles in the internodal region and a relatively high concentration at the node of Ranvier (approximately 1200 particles mum-2). The nodal particles tend to be larger than most intramembranous particles, approaching 200 A diameter. The paranodal region of the leaflet is virtually devoid of such particles except in the narrow helical 'groove' which faces extracellular clefts between terminating glial processes. In places this pathway widens to form 'lakes' up to approximately 0.3 mum2 area which contain large numbers of large particles resembling those at the node. The concentration of particles at the node is in the same range as the concentration of sodium channels estimated to be in this region and it is suggested on the basis of their location and concentration that these particles represent ionophores. The distribution of particles in the paranodal region suggests that the large intramembranous particles do not have free access to the axoglial junctional portion of the membrane and therefore the movement of such particles along the paranodal region of the membrane may occur primarily in the membrane of the 'groove' spiraling through this portion of the axolemma. Such a restriction in surface area for particle movements on either side of the node of Ranvier could result in trapping of particles at the node and thus contribute to their concentration in the nodal axolemma.

Animals↗