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C Ide

Publications and source records attributed to C Ide.

At least 73 records · Page 4Linked to original sources

Distribution of protein kinase C (alpha, beta, gamma subtypes) in normal nerve fibers and in regenerating growth cones of the rat peripheral nervous system.

The distribution of protein kinase C (alpha, beta, gamma subtypes) was studied using immunocytochemical techniques in normal nerve fibers and in regenerating sprouts (growth cones) from the nodes of Ranvier following crush injuries to the rat peripheral nervous system. In normal nerves, for each protein kinase C subtype, immunoreactivity was present in both myelinated and unmyelinated axons. In myelinated axons, immunoreactivity for all three subtypes was patchy in the axoplasm and diffuse in the subaxolemmal peripheral zones. No immunoreactivity was found in the microtubule and neurofilament (cytoskeletal) domain. In contrast, in unmyelinated axons, immunoreactivity was distributed diffusely in the axoplasm. Schwann cells of myelinated fibers exhibited protein kinase C immunoreactivity, but those of unmyelinated fibers did not. In regenerating nerves, early sprouts and growth cones extending through the crushed site along Schwann cell basal laminae exhibited intense immunoreactivity for all three subtypes. Immunoreactivity was distributed diffusely throughout the axoplasm of the regenerating sprouts (growth cones), in which microtubules and neurofilaments were very rare. Thus, the subcellular localization of the protein kinase C immunoreactivity in growth cones of early regenerating nerves differed from that of normal parent axons. These findings suggest that protein kinase C (alpha, beta and gamma subtypes), whose subcellular distribution becomes more extensive in regenerating axons, may have important functional roles in axonal sprouting and in the regulation of growth cone activity in the peripheral nervous system.

Animals↗

Localization of N-cadherin in the normal and regenerating nerve fibers of the chicken peripheral nervous system.

The localization of N-cadherin in the normal, and regenerating nerve fibers was investigated by immunocytochemistry in the chicken sciatic nerve. The normal unmyelinated fibers exhibited N-cadherin immunoreactivity on the plasma membranes of axons and Schwann cells where they were in contact with each other, while myelinated fibers displayed no immunoreactivity except at the mesaxon where Schwann cell plasma membranes were attached to each other. In the regenerating nerves, intense immunoreactivity was demonstrated on the surface of plasma membranes of axons and Schwann cells where axon-axon and axon-Schwann cell contacts were made. No immunoreactivity was observed on the plasma membranes where regenerating axons or Schwann cells were in touch with the basal lamina. In addition, it was revealed that some vesicles in the growth cones had distinct N-cadherin immunoreactivity at the inner limiting membrane surface. These findings indicate that N-cadherin may be involved in the axon-axon and axon-Schwann cell adhesion in the normal unmyelinated as well as regenerating nerve fibers, and also in the attachment of Schwann cell processes at the mesaxon of myelinated fibers. In addition, these findings suggest that N-cadherin might be, at least in part, supplied by fusion of growth cone vesicles with the surface plasma membranes in growing axons.

Animals↗

Localization of Rabphilin-3A on the synaptic vesicle.

Rabphilin-3A is a putative target protein for Rab3A small GTP-binding protein which is implicated in neurotransmitter release. Rabphilin-3A is expressed mainly in brain, but its subcellular localization remains to be clarified. Immunohistochemical analysis has revealed that Rabphilin-3A is most abundant in the synaptic area of the rat cerebellum, retina, and neuromuscular junction. Ultrastructural analysis of the neuromuscular junction using the immunogold method indicates that Rabphilin-3A is localized on the synaptic vesicle. Subcellular fractionation analysis of rat brain has shown that Rabphilin-3A is most highly concentrated in the purified synaptic vesicle fraction. These results indicate that Rabphilin-3A is localized on the synaptic vesicle in the presynapse.

Adaptor Proteins, Signal Transducing↗

Localization of rabphilin-3A, a putative target protein for Rab3A, at the sites of Ca(2+)-dependent exocytosis in PC12 cells.

Rab3A/Smg 25A, a small GTP-binding protein, is highly concentrated in presynapse of neurons and implicated in neurotransmitter release. We have recently identified a putative target protein for Rab3A, isolated its cDNA, and designated it as Rabphilin-3A. To examine whether Rabphilin-3A as well as Rab3A is localized at the sites of Ca(2+)-dependent exocytosis, we investigated here localization of Rabphilin-3A and Rab3A in comparison with the sites of exocytosis in the differentiated PC12 cells. Rabphilin-3A as well as Rab3A was highly concentrated at the tips of the neurites where Ca(2+)-dependent exocytosis took place. Inversely, neither Rabphilin-3A nor Rab3A was concentrated at the tips of the neurites where Ca(2+)-dependent exocytosis did not take place. These results suggest that Rabphilin-3A as well as Rab3A constitutes a part of the machinery necessary for neurotransmitter release.

Adaptor Proteins, Signal Transducing↗

Localization of protein kinase C alpha, beta and gamma subspecies in sensory axon terminals of the rat muscle spindle.

The localization of protein kinase C (PKC) alpha, beta and gamma subspecies in sensory axon terminals of muscle spindles in the plantar lumbrical muscles of rat was investigated by light and electron microscopic immunocytochemistry using monoclonal and polyclonal antibodies. Immunoreactivity for these subspecies was detected specifically in sensory axon terminals which wound spirally around the intrafusal muscle fibres of the muscle spindle. Immunostaining was found to be stronger with polyclonal than with monoclonal antibodies. By electron microscopy, immunoreactivity for alpha, beta and gamma subspecies was almost diffusely distributed in the cytoplasm of the axon terminal, and the overall pattern of distribution of immunoreactivity was similar for all three subspecies. In the cases of alpha and beta subspecies, some intensely immunostained regions were found in the cytoplasm, but no definite subcellular structures corresponding to such regions could be identified. Considering that PKC plays a crucial role in the regulation of ion channels, it is suggested that PKC might be involved in the control of mechanoelectric transduction in sensory axon terminals.

Animals↗

Experimental syringomyelia in the rabbit: an ultrastructural study of the spinal cord tissue.

Hydrosyringomyelia was produced experimentally by the injection of kaolin into the cisterna magna of the rabbit, and the ultrastructural changes of the spinal cord surrounding the syrinx were investigated 2, 4, and 6 weeks after injection by transmission electron microscopy. The ependyma at the ventral part of the central canal was flat and stretched, whereas, in the dorsal part, it was split, and the syrinx extended through the dorsal median plane in most animals. Extracellular edema was found in the subependymal white matter and in and around the posterior median septum. Many nerve fibers surrounding the syrinx were in varying stages of axonal degeneration. Myelin sheaths were split, thinned, and completely lost in many nerve fibers. In some fibers, the axons were totally lost, leaving the myelin sheaths as empty tubes. Astrocytic processes containing a large number of glial filaments covered the nerve fibers adjacent to the syrinx and partially replaced the edematous area. The perivascular spaces were enlarged, especially near the syrinx and in the dorsal white matter. Oligodendrocytes remained undamaged, and the remyelination by oligodendrocytic processes was seen on some denuded axons. Sometimes, this further remyelination was abortive, especially where the edema was severe. The ultrastructural changes of the neural tissue and their sequences were identical, in most respects, to those of hydrocephalus and noncommunicating syringomyelia. The oligodendrocytic remyelination with ongoing demyelination found in this model has many similarities to those in experimental hydrocephalus.

Animals↗

Motor axon terminal regeneration as studied by protein gene product 9.5 immunohistochemistry in the rat.

Normal intact and regenerating axon terminals up to 30 days after nerve crushing were studied in the rat flexor carpi ulnaris muscle by confocal laser scanning microscopy (CLSM) and electron microscopy using immunohistochemistry for protein gene product 9.5 (PGP 9.5). The motor axons were intensely and homogeneously stained along their entire length. "Three dimensional" organizations of elaborate axon terminals were clearly demonstrated by reconstructing serial optical images obtained by CLSM in normal neuromuscular junctions. In the injured nerve, the earliest regenerating axons could be identified at endplate regions six days after nerve crushing as intensely immunoreactive thin processes which bifurcated in T-shape and formed delicate lace-like terminals. Such lace-like terminals were composed of fine thread-like portions 0.2-0.8 microm in diameter and expanded portions of 2-3 microm in diameter. Electron microscopy revealed that all the axon terminals in the cytoplasm were stained almost homogeneously by PGP 9.5 immunohistochemistry up to their extreme tips. Axon terminals were in direct contact with the basal lamina of the postsynaptic folds, and showed occasional branching. The thin thread-like portions contained no mitochondria but only a few vesicles, where as the expanded portions, abundant mitochondria. And preterminal axons and some expanded portions were abutted by Schwann cells, while thin thread-like portions were exposed with no association with Schwann cells. Twenty to 30 days after crushing injury, regenerating motor axon terminals resumed their mature form in terms of branching elaborations and ultrastructural features. Thus, CLSM of PGP 9.5 immunocytochemistry combined with electron microscopy was able to demonstrate the "three-dimensional" organization of elaborate axon terminals at high resolution in the normal and regenerating neuromuscular junctions. Using this technique, extremely fine processes of axonal terminals were identifiable at the earliest stage of reinnervation.

Animals↗

[Diagnosis of acute colonic diverticulitis: comparison with echography and tomodensitometry].

The sensitivity of ultrasonography (US) as the initial imaging procedure in acute diverticulitis of the left colon was retrospectively compared with that of computed tomography (CT) in 34 patients. Final diagnosis was based on clinical findings and follow-up in a first group of 20 patients with mild diverticulitis, and was proved by surgery in a second group of 14 patients with severe diverticulitis. In the first group, US showed findings consistent with diverticulitis in 19 patients, as did CT. The image of an inflamed diverticulum was shown by US in 10 patients. In the surgical group, findings consistent with diverticulitis were shown by US in 11 cases, and by CT in 13 cases. An inflamed diverticulum was not detected at US in this group of patients, presumably because the diverticulum was incorporated in the inflammatory process. These results suggest that US is a valuable technique in the initial evaluation of patients suspected of having acute diverticulitis, especially when the disease is not severe. The image of an inflamed diverticulum shown at US strongly suggests the diagnosis of mild diverticular disease. When diverticulitis is severe, CT should be performed in addition to US.

Acute Disease↗

Synaptophysin immunocytochemistry in the regenerating sprouts from the nodes of Ranvier in injured rat sciatic nerve.

Following crush injury of rat sciatic nerve, strong synaptophysin immunoreactivity was demonstrated in the regenerating sprouts that emerged from the proximal nodes of Ranvier and in their growth cones that extended through the space between Schwann cell basal lamina and myelin sheath of the parent axon. These findings suggest that synaptophysin is involved in the growth regulation of regenerating sprouts.

Animals↗

Ultrastructural localization of protein kinase C beta-subspecies in the axon terminal of rat neuromuscular junction.

Ultrastructural localization of protein kinase C (PKC) beta-subspecies in neuromuscular junctions of the rat lumbrical muscle was investigated by the immunoperoxidase and immunofluorescence methods. By light microscopy, PKC beta-like immunoreactivity (PKC beta-LIR) was found in the axon terminal expansions as well as in the preterminal axons. By confocal laser scanning microscopy, the staining for PKC beta-like immunoreactivity was more intense in the presynaptic regions just in contact with the acetylcholine receptor stained by FITC-alpha-bungarotoxin. By electron microscopy, PKC beta-like immunoreactivity was distributed non-uniformly in the terminal expansions. In the terminal expansions, PKC beta-like immunoreactivity was accumulated in the presynaptic regions in contact with the post-synaptic folds. This accumulation was approximately 0.1-0.2 microns in diameter, which comprised a part of the presynaptic plasma membrane and a group of synaptic vesicles adjacent to it. Weak immunoreactivity was also found diffusely in the axoplasmic matrix. The discrete presynaptic accumulation of PKC beta-subspecies may represent the strategical localization specialized for the effective regulation of neurotransmitter release.

Animals↗

Localization of smg p25A/rab3A p25, a small GTP-binding protein, at the active zone of the rat neuromuscular junction.

smg p25A is a small G protein which has been suggested to regulate neurotransmitter release from the synapses. We investigated here the ultrastructural localization of this small G protein in the rat neuromuscular junction by an immunoperoxidase method. The results showed that smg p25A was distributed non-uniformly on the presynaptic plasma membrane and among the synaptic vesicles with the focal accumulation on the discrete presynaptic sites which corresponded to the active zones, the regions of the presynaptic plasma membrane specialized for the exocytosis of the synaptic vesicles. This unique distribution of smg p25A suggests that it plays an important role in the attachment and fusion of the synaptic vesicles with the active zones.

Animals↗

Protein kinase C (alpha, beta, gamma) in Pacinian corpuscle.

Immunocytochemical demonstration of protein kinase C (PKC) subspecies (alpha, beta, gamma) was carried out in Pacinian corpuscles of rat hind feet using monoclonal or polyclonal antibodies against each of these subspecies. The inner core cells and lamellae and the Schwann cell cytoplasm of the nerve fiber innervating the corpuscle were strongly positive for PKC alpha-immunoreactivity (IR). In contrast, the axon terminal and the outer core did not display any positive alpha-IR. Very weak PKC beta-IR was detected in the ultraterminal region of the axon terminal, while the trunk region showed no immunoreactivity. Very faint PKC beta-IR was found also in the lamellar cells located at the periphery of the inner core and the endoneurial fibroblasts in the intermediate layer. PKC gamma-IR was not detected in any part of the corpuscle. The strong PKC alpha-IR in the inner core and the presence or absence of PKC alpha-, beta-, and gamma-IR in the axon terminal are discussed from the point of view of the functional aspects of each part.

Animals↗

Traumatic degeneration of transected myelinated fibers of the mouse sciatic nerve.

Traumatic degeneration of myelinated fibers was studied by electron microscopy over 5 days following transection of mouse sciatic nerve. Special attention was paid to the mechanism which separates the degenerating part, while preserving the viable part of the axon. Immediately after transection, the opened end of the proximal stump revealed extensive subcellular changes including the disorganization of neurofilaments, and disruption of mitochondria and axonal endoplasmic reticulum (SER). Subsequently, vesicles of round and tubular profiles filled up the whole area of the stump end, and proximal to it appeared a neurofilament-predominant area characterized by randomly oriented neurofilaments and normally appearing mitochondria and SER. Characteristic membranous demarcations occurred in early periods at the border between the vesicle accumulation and the neurofilament-predominant areas, and later also within these areas. The demarcation membranes formed both by invagination of the surface plasma membrane and, probably, by fusion of the large vesicles. These became prominent with time, dividing the axoplasm into compartments of varying sizes, which gradually underwent degeneration and were liberated from the parent axon. Occurrence of autophagic vacuoles was characteristic of the degenerating portions of the parent axon. Thus, by the function of demarcation membranes, the parent axon to be preserved could remain membrane-bound, while the degenerating parts were shed off.

Animals↗

Aberrant remyelination of axons after heat injury in the dorsal funiculus of rat spinal cord.

We studied the course of demyelination and subsequent remyelination of nerve fibers after heat injury in the dorsal funiculus of the rat spinal cord. Four weeks after heat treatment, we observed, in addition to normally remyelinated axons, a few aberrantly remyelinated axons which had both CNS- and PNS-type myelin sheaths: the CNS-type myelin sheaths were always situated inside the PNS-type sheaths. This finding indicates that in some conditions Schwann cells can form myelin sheaths around those formed by oligodendrocytes.

Animals↗

Ultrastructure of the nerve endings in the rat tongue mucosa.

Ultrastructure of the sensory nerve endings in the dorsal tongue mucosa of Sprague Dawley rats was described. Thick bundles of nerve endings, the presence of branches and organized or free nerve endings were found in the subepithelial space. Lamellar corpuscles containing several terminal axons were found in close contact with basal lamina of the epithelium. The lamellar cells envelope the axons at different levels. The cytoplasm of lamellae contains numerous microfilaments, microtubules, mitochondria and caveolae. The terminal axons measure 0.5 to 1.0 micron in diameter and present several forms. Short axoplasmic protrusions and dilated ends were observed. The axons contain numerous mitochondria, microtubules, neurofilaments and small clear vesicles.

Actin Cytoskeleton↗

Regeneration through nerve allografts in the cynomolgus monkey (Macaca fascicularis).

With the use of ulnar nerves of cynomolgus monkeys, the present study examined whether basal laminae of Schwann cells can serve as conduits for regenerating axons in nerve allografts from non-human primates. A segment of ulnar nerve was transected distal to the elbow joint one week before grafting. In Group A, a distal segment of the transected nerve was transplanted, after freezing and thawing, into the ulnar nerve of another monkey, at a level that corresponded to that from which the graft was taken. In Group B (the control group), the segment of nerve was grafted in the same manner but without cryotreatment. Two weeks, five weeks, eight weeks, and five months after grafting, the graft and the host nerve were examined with light and electron microscopy. Within two weeks after grafting in Group A, after degradation of the cellular components of the Schwann cells, the basal laminae of the Schwann cells were intact in the form of tubes. Within five weeks, many regenerating axons grew out into these basal lamina tubes in the three-centimeter-long grafts and extended into the host nerve. As seen at the wrist (seven centimeters from the distal suture) five months after grafting, the axons exhibited fully mature myelination both in the graft and in the host nerve. In contrast, in Group B, in which the Schwann cells had not been disrupted by cryotreatment, cellular components and connective-tissue matrices, including basal laminae, had been degraded and had been replaced by invading cells, which filled the endoneurial spaces of the graft. Five months after grafting, axonal growth had been arrested in the graft one centimeter distal to the proximal suture. The beneficial effect in Group A appears to have been the result of the retention and preservation of intact basal laminae of Schwann cells after rapid removal of killed Schwann cells and myelin debris. Killing of Schwann cells by freezing before grafting may abolish the immune response to the Schwann cells in allografts and lead to fragmentation and disruption of myelin, which facilitates the rapid removal of myelin by macrophages.

Animals↗

An experimental study of nerve regeneration through chemically treated allografts.

We carried out experiments in rabbits to determine whether treating nerve transplants with gradually increasing concentrations of ethanol, ether and ficin would inhibit the graft-host immune reaction to the allograft. After treatment with ethanol, the basal laminar scaffold of the Schwann cell remained intact and there was satisfactory axonal regeneration. The results after additional treatment with ether or ficin did not achieve such good results. Preservation of the basal lamina is considered to be the essential factor in allowing neural regeneration in these circumstances.

Animals↗