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C Ide

Publications and source records attributed to C Ide.

At least 109 records · Page 6Linked to original sources

A modified KOH-collagenase method applied to scanning electron microscopic observations of peripheral nerves.

Certain parts of the peripheral nervous system were observed by a modification of the KOH-collagenase method by Miller et al. (1982). The sciatic nerve of the mouse, and dorsal root ganglion, lingual muscle and jejunum of the rat were fixed over one day at room temperature in a fixative containing paraformaldehyde and glutaraldehyde. After fixation, specimens were treated with 5N KOH solution for 5-10 min at 60 degrees C, immersed in a collagenase solution for 3-5 h at 37 degrees C and processed for scanning electron microscopy (SEM). In adequately treated specimens, connective tissue matrices and basal laminae were completely removed without causing any severe tissue damage. The three-dimensional visualization of cellular elements of peripheral nerves was enabled with the following results: 1) Individual nerve fibers of the sciatic nerve were clearly exposed. Myelinated fibers were non-branching cords with local annular constrictions at the node of Ranvier, while unmyelinated ones branched and anastomosed with one another to form loose networks. The sites of accumulation of the Schwann cell cytoplasm swelled on the surface of the myelinated fibers. Mesaxons were visualized as longitudinal furrows. 2) Ganglion cells covered by satellite cells were observed in the dorsal root ganglion. The ganglion cells were covered by their own convoluted dendro-axonal processes, thus forming the initial glomeruli of Cajal. Satellite cells at the glomeruli extended many finger-like projections surrounding the dendro-axonal processes. 3) Motor endplates were observed in lingual muscles. Terminal Schwann cells (teloglia) at the endplate were clearly visualized: the round perikaryon extended cytoplasmic processes along the axonal branches within the endplate. The processes issued fine finger-like projections from their margins. 4) Vascular autonomic plexuses were clearly demonstrated in lingual muscles. Unmyelinated nerves branched and anastomosed to form elaborate nerve networks around the vessels. Neuronal processes and associated Schwann cells were identifiable at high magnification. 5) Submucous nerve plexuses in the jejunum consisted of numbers of ganglia and interconnecting strands of fibers which formed very complicated networks. These observations indicate that this modified KOH-digestion method is useful for the SEM study of the three-dimensional cellular organization of nervous elements in various tissues.

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Autonomic nerve networks in the rat exocrine pancreas as revealed by scanning and transmission electron microscopy.

The three-dimensional architecture of the autonomic nerve terminals in the rat exocrine pancreas was investigated by scanning electron microscopy using the HCl-digestion method as well as by transmission electron microscopy. Unmyelinated nerves presumed to be autonomic in nature were found in networks covering the outer layers of arterioles and their capillary extensions, with nerve fibers often leaving the capillaries to surround acini in the interacinar spaces. Schwann cells formed scaffolds for axons of the networks. No other distinct type of cells such as the so-called interstitial cells of Cajal were found to be associated with the formation of the networks. Although nerve fibers of the networks were locally in close association with the walls of the blood vessels or with the bases of acinar cells, no specialized axonal contacts with these tissues were found. However, local swellings, presumably varicosities, were observed by scanning electron microscopy on the surface of nerves. These findings suggest that the networks of unmyelinated nerves represent terminal apparatuses of the autonomic nerves in the pancreas. Schwann cells in the terminal networks were characterized by an unusual abundance of cell organelles. These "terminal Schwann cells" well correspond in location and reticular extension to the "interstitial cells of Cajal" as demonstrated by silver impregnation and vital methylene blue staining. The occurrence of well developed Golgi apparatuses, rough endoplasmic reticulum, and numerous ribosomes suggests that the cells are specialized Schwann cells which most likely require high levels of cellular activity in order to maintain their elaborate cytoplasmic processes extending along the terminal networks, and also to sustain the specific functions of axon terminals.

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Nerve regeneration in cornea after penetrating keratoplasty in rabbit, with special reference to relationship between regenerating nerves and basal laminae.

The pattern of nerve regeneration in the grafted rabbit cornea was investigated by electron microscopy. Grafted corneas were excised 2, 7, 14 and 28 days after grafting, and processed for observation by conventional electron microscopy. In the normal, unoperated cornea Schwann cell basal laminae are, unlike those of ordinary peripheral nerves, discontinuous and fragmentary on the fibers coursing through the corneal stroma. In the early stage of regeneration, while numbers of regenerating axons extended through the Schwann cell columns of regenerating axons extended through the Schwann cell columns in the grafts, many other regenerating axons elongated as single fibers through the corneal stroma outside the Schwann cell columns. These single naked axons were later enveloped by Schwann cell cytoplasm, contributing to the overall dense irregular pattern of regenerated nerves in the grafted cornea. It was thought that the regenerating axons can extend throughout the stroma without the guidance of basal lamina tubes, making the corneal stroma a favorable environment for nerve regeneration.

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Role of extracellular matrix in the regeneration of a pacinian corpuscle.

The pacinian corpuscle is composed of an inner and an outer core or bulb. The former is formed by tightly packed and multi-layered thin cellular processes (lamellae) of lamellar cells which surround a centrally located axon terminal, and the latter, also called the capsule, is made up of very loosely piled layers of thin cells which encircle the inner core. Lamellar cells of the inner core are considered to be specialized Schwann cells, and the outer core cells are modified perineurial cells. In the present study, the matrix filling the extracellular spaces of the inner core consisted of basal lamina-like amorphous materials, sparce fine collagen fibrils, and the ground substance embedding these structural components. No definite basal laminae were found on the inner core lamellae except on the peripherally located ones which had distinct basal laminae. Outer-core cells were invested along the entire contour by distinct basal laminae. The interspace between the inner and outer cores was a continuation of the nerve endoneurium. The purpose of this investigation was to determine whether the extracellular matrix of the pacinian corpuscle, especially that of the inner core, has the ability to cause corpuscle regeneration, i.e. to make the regenerating axons and Schwann cells differentiate into corpuscular axon terminals and inner core cells, respectively. Pacinian corpuscles in the periosteum at the distal end of the fibula of mice were repeatedly frozen (3-5 times) in situ with forceps cooled with liquid nitrogen. Within 2-3 days, all the cellular constituents of the corpuscle had degraded, while the extracellular matrices of the inner and outer cores apparently remained undamaged. After 5-7 days, regenerating axons and accompanying immature Schwann cells entered these extracellular matrices of the inner cores. A remarkable finding was that these immature Schwann cells were detached from the axon, and sent thin cellular processes around the axon in a characteristic fashion, basically forming the same pattern as lamellae in a normal corpuscle. The regeneration of the inner core was completed by about 40 days after the freezing treatment. In the outer core, perineurial cells proliferated and extended through the basal lamina tubes of the old cells, becoming new outer core cells. These findings indicate that the extracellular matrix of the pacinian corpuscle has a specific property to cause the regeneration of the corpuscle.

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Scanning electron microscopic studies of the myelinated nerve fibres of the mouse sciatic nerve with special reference to the Schwann cell cytoplasmic network external to the myelin sheath.

The three-dimensional morphology of the surface of myelinated nerve fibres in the mouse sciatic nerve was studied by scanning electron microscopy after combined potassium hydroxide treatment and collagenase digestion (to remove the surrounding collagen fibrils and basal laminae from nerve fibres) as well as by transmission electron microscopy. The myelinated nerve fibre appeared as a long cylinder with sporadic annular constrictions corresponding to the nodes of Ranvier. Slight swellings of the surface due to Schwann cell nuclei were usually found at the middle of each internode. The surface of the nerve fibre clearly exhibited a network of bulges, which consisted of longitudinal bands extending from the nuclear swelling to the nodes of Ranvier through the internode, and transverse trabeculae bridging between these longitudinal bands. These bulges on the surface of nerve fibres were the site of the retained Schwann cell cytoplasm external to the myelin lamellae. These cytoplasmic networks on myelinated fibres presumably corresponded to the networks described by Cajal following silver impregnation. In addition, other thin elevations and focal round swellings were also found associated with these longitudinal bands and transverse trabeculae. These networks of Schwann cell cytoplasm are considered to be cytoplasmic channels for nutrition. The two apposing paranodal bulbs of nodes of Ranvier were often asymmetrical in their structure. The networks of the paranodal region were more complicated than those in the internode. The networks of Schwann cell cytoplasm converged into a continuous circumferential collar toward the node, which in turn gave rise to finger-like projections into the nodal gap.

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Specializations of plasma membranes in Pacinian corpuscles: implications for mechano-electric transduction.

Pacinian corpuscles of cat mesentery were studied with freeze-fracture and thin sectioning methods after chemical fixation. Intramembranous particles (IMPs) exhibit differences in both density and pattern of distribution between the axolemma of the smooth short axis (x-axis) region and that of the axonal spine region of the long axis (y-axis) of the axon terminal. The axolemma of the x-axis has IMPs at a density of 2687 +/- 581 per micron2 (mean +/- S.E.M.), and these particles are 9.0 +/- 1.7 nm (mean +/- S.D.) in diameter. In contrast, the axolemma of the y-axis has a higher density of IMPs (3607 +/- 612 per micron2) which are larger (diameter, 10.0 +/- 1.7 nm). The particle distribution is not homogeneous in x-axis membranes as there are small patchy areas devoid of particles scattered throughout the entire surface. The E-face of the axolemma has a low density of IMPs (approximately 200 per micron2 in both x- and y-axes). However, IMPs in the E-face are smaller (approximately 9 nm) in the x-axis than in the y-axis (approximately 10 nm). The inner core lamellar cells have IMPs at a density of 3276 +/- 739 per micron2 and 553 +/- 169 per micron2 in the P- and E-faces, respectively. The particles are about 10 nm in diameter in both faces. Many gap junctions occur between lamellar cells especially near the clefts, suggesting that hemilamellae of each inner core half are kept at the same electrotonic potential. The outer core lamellar cells have IMPs at a density of 2239 +/- 403 per micron2 and 536 +/- 123 per micron2 in their P- and E-faces, respectively. The particles are approximately 10 nm in diameter in both faces. A noteworthy finding is that tight junctions are prominent at cell-to-cell appositions within individual lamellae, especially in the first and second (or sometimes third) innermost lamellae of the outer core. These tight junctions are considered to be a barrier to the leakage of fluid and/or ions between interlamellar spaces as well as between inner and outer cores. An intermediate cell layer is identified between the inner and outer cores. The connective tissue space of this cell layer corresponds to the endoneurium, indicating that intermediate layer cells are comparable to endoneurial fibroblasts.(ABSTRACT TRUNCATED AT 400 WORDS)

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Cholinesterase activity of lamellated sensory corpuscles in the rat lip.

Non-specific cholinesterase (ChE) activity was demonstrated in lamellated sensory corpuscles of the rat lip by light and electron microscopy using Karnovsky and Roots' method. ChE activity was present in the interlamellar spaces between neighbouring lamellae as well as in the peri-axonal space between axon terminals and their adjacent lamellae. Reaction products were also deposited in some caveolae of the lamellar cell plasma membrane, and in the cisternae of the rough endoplasmic reticulum as well as in the nuclear envelope of lamellar cell bodies. No reaction products were detected within the axon terminals. The findings show that the lamellated corpuscles in the rat lip, like other mechanoreceptors, have intense ChE activity which is probably synthesized in lamellar cells and released into the intercellular spaces. That ChE activity is particularly strong in mechanoreceptors composed of lamellar cells suggests that this enzyme would play an important role in function and/or maintenance of such mechanoreceptive corpuscles.

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The enigma of sensitivity in Pacinian corpuscles: a critical review and hypothesis of mechano-electric transduction.

The present report reviews the physiological and morphological specializations of Pacinian corpuscles and other mechanoreceptors that are present in the skin and connective tissues of the body as well as the cochlea. The remarkable sensitivity of Pacinian corpuscles is such that the only form of mechanical energy that could be perceived by a Pacinian corpuscle is a sound wave. In fact the human finger as demonstrated by Munger and Ide (1987) can perceive sound waves when water is the coupling agent. The structural specializations are equally remarkable with extensive membrane specializations of both the inner core and inner portion of the outer core. The halves of the inner core are each coupled with gap junctions and the inner portion of the outer core joined with numerous tight junctions. The cleft regions have specializations involving the axolemma that consist of numerous axonal spines containing bundles of filaments projecting into the cleft of the inner core. These structural specializations are thought to represent specializations for mechano-electric transduction analogous in many respects to the hair cells of the cochlea. A hypothesis for mechano-electric transduction is presented that may account in part for the extreme sensitivity of Pacinian corpuscles and other mechanoreceptors.

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Nerve regeneration through cryo-treated xenogeneic nerve grafts.

Cryo-treated nerves whose Schwann cells had been killed by repeated freezing and thawing were xenogenically grafted into sciatic nerves from rats (Wistar, as donor) to mice (ddy strain, as recipient) to examine whether Schwann cell basal lamina tubes of cryo-treated xenogeneic grafts were effective conduits for regenerating axons. For comparison and evaluation of the effectiveness of this technique, experiments using grafts without the cryo-treatment were carried out. Cells in cryo-treated xenografts degraded into cell debris immediately after grafting and then were phagocytized by macrophages. After the cellular components had been removed from the graft, Schwann cell basal laminae remained intact in situ, serving as conduits for the regenerating axons. The process of nerve regeneration was almost the same as that observed in cryo-treated auto- and allografts, except that the regeneration was slightly delayed in the xenogeneic graft. In contrast, an extensive cell infiltration occurred in the non-treated grafts. It appeared that the donors Schwann cells in the graft deteriorated due to immunological reactions and were finally eliminated by macrophages, leaving their basal laminae undamaged in situ. The initiation of nerve regeneration including perineurial sheath formation in non-treated grafts was, therefore, significantly delayed, but once begun, it proceeded in the same manner as in the cryo-treated grafts. These findings strongly indicate that Schwann cell basal laminae can serve as effective pathways for regenerating axons even in the xenograft. Moreover, cryo-treated xenogeneic grafts are more desirable than non-treated ones, since dead Schwann cells in the former can be removed in the early period (4-14 days) from the graft without causing any immunological reaction, thus resulting in the facilitation of nerve regeneration.

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Carbonic anhydrase activity in axon terminals of sensory corpuscles.

The distribution of carbonic anhydrase (CA) activity was studied by electron microscopic histochemistry in rat Pacinian corpuscles, Meissner corpuscles and Merkel cell-neurite complexes using the cobalt bicarbonate method. The distribution of CA activity in these axon terminals was compared to the activity in sciatic nerve axons. An intense enzymatic CA activity was demonstrated in axon terminals of both Pacinian and Meissner corpuscles, while a weak activity was found within the axoplasm of terminals abutting Merkel cells. Some large- and medium-sized axons in sciatic nerves exhibited an intense activity. These findings indicate that large- or medium-diameter sensory axons innervating corpuscular endings have an intense CA activity extending from their somata to their sensory terminals. Axons to Merkel-neurite complexes differ in CA activity from those innervating Meissner and Pacinian corpuscular endings.

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The cytology of human Pacinian corpuscles: evidence for sprouting of the central axon.

During the course of the studies on non-traumatized Pacinian corpuscles from normal human adults, we have frequently encountered corpuscles which have an 'apparently multiple' innervation in both light and electron microscopic preparations. On closer inspection of serial sections for both light and electron microscopy, these 'apparently multiple' axon terminals have been found in fact to be branches of the main central axon within the inner core of the corpuscle. Sprouting occurred at the trunk or at the extreme tip of the main axon, and such sprouts extended in various directions from the central axon throughout the inner core, producing tortuous and complex patterns of this 'multiple' innervation. These axonal sprouts do not have separate inner cores separated from one another, but rather are embedded in a common inner core. The presence of a common inner core thus differentiates normal axonal sprouts from the experimentally or pathologically produced multiple innervation that results form regeneration of axons in a previously denervated corpuscle. We conclude that the inner core of Pacinian corpuscles is a unique micro-environment promoting sprouting of sensory axon in the normal human adult as well as juvenile Pacinian corpuscles.

Adolescent↗

Basal laminae and Meissner corpuscle regeneration.

Murine Meissner corpuscles (mouse digital corpuscles), located in pad skin at the toe tip, consist of lamellar cells with long cellular processes (lamellae) surrounding axon terminals in an onion-skin fashion. Lamellar cell bodies and processes were provided with a basal lamina. The present study was made to examine whether these lamellar cell basal laminae have any specific role in the differentiation of regenerating axons and Schwann cells into specialized axon terminals and lamellar cells, respectively. Pad skin at the toe tip was treated 3-5 X by freezing and thawing. By this treatment, cellular constituents of the corpuscles die and disintegrate into cell debris, leaving in situ basal laminae of the lamellar cells in stacked hollow loops, reminiscent of the original configuration of lamellae. Schwann cells and axons of the ordinary nerve fibers in the pad skin were similarly damaged, and basal laminae of the Schwann cells remained as basal lamina tubes. Three days after treatment, regenerating axons were seen extending through the basal lamina tubes of Schwann cells deep in the toe pad skin. However, no regenerating axons were found in the vicinity of the old corpuscles. Five days after treatment, regenerating axons, some of which were accompanied by migrating Schwann cells and others which were still naked, were noted at the subepidermal region, and began to enter the hollow basal lamina loops of the old corpuscles. Eight-15 days after treatment, regenerating axons which entered the basal lamina loops successively gave rise to branches, and at the same time, accompanying Schwann cells emanated cellular processes through well-preserved basal lamina loops. Fifteen-25 days after treatment, regenerating axons seemed to be morphologically specialized as axon terminals, and accompanying Schwann cells differentiated into definite lamellar cells which surrounded the axon terminals in the same manner as in the normal murine Meissner corpuscles. Although the incidence of good regeneration of the corpuscle was relatively low, these findings suggested that basal laminae of lamellar cells might have some specific properties which could be responsible for the differentiation as well as maintenance of lamellar cells and axon terminals in the Meissner corpuscles.

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Changes in thickness of collagen fibrils in the endo- and epineurium of the mouse sciatic nerve during development.

Changes in the diameter of collagen fibrils were observed with an electron microscope in the endo- and epineurium of sciatic nerves of mice during development from 12 days of gestation to 5 months after birth. It was noted that endoneurial collagen fibrils appeared in embryonic mice at 15 days of gestation, and at the same time, basal laminae began to appear sporadically on the Schwann cell plasmalemma. No fibroblasts were seen at this developmental stage. Collagen fibrils in the endoneurium remained as thin as they were when they first appeared, being in the narrow range of 250-300 A in diameter, while those in the epineurium became much thicker (400-450 A, 5 months after birth) as is also the case in dermal connective tissues. The present study shows that the endoneurial collagen fibrils were different in their developmental pattern from those of the epineurial or of other connective tissues, lending support to the concept that the endoneurial collagen fibrils are particular in nature, being so-called histological reticular fibers.

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[Ultrastructural study of the ureter smooth muscle of the cat].

Morphological specializations of smooth muscles from the middle part of the cat ureter have been examined by electron microscopy using thin sections and freeze-fracture replicas. It is notable that there were many gap junctions which were characteristic in being formed on the surface of blunt cytoplasmic projections protruded from each neighboring cells. In addition to gap junctions, there were two other different junctions, i.e, intermediate junctions and "focal attachments". The latter represents the cell attachment in which two apposing plasma membranes are attached to each other without the intervension of basal lamina in the narrow intercellular space. Caveolae were lined in rows which were intervened by dense band zones parallel to the long axis of the muscle fibre as seen by freeze-fracture replica. Gap junctions were noted to be localized within or along the caveolar zones, being elliptical in shape with the long axis parallel to muscle fibres. As observed in thin sections, gap junctions in freeze-fracture replicas were noted on the top of cytoplasmic protrusions. No membrane specializations were observed in freeze-fracture replicas which corresponded to the "focal attachment" or intermediate junction as seen in thin sections. As to the innervation, the autonomic nerve terminals were in abundance, closely apposed to the smooth muscles. In addition to autonomic nerve terminals, sensory nerve terminals were observed between muscle fibres, consisting of axon terminals and lamellar cells, the same structure as mechanoreceptive lamellar corpuscles known in other organs.

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Cholinesterase activity of lamellated sensory corpuscles in the rat lip.

Non-specific cholinesterase (ChE) activity was demonstrated in lamellated sensory corpuscles of the rat lip by light and electron microscopy using Karnovsky and Root's method. ChE activity was present in the interlamellar spaces between neighbouring lamellae as well as in the periaxonal space between axon terminals and their adjacent lamellae. Reaction products of ChE activity were also deposited in some caveolae of the lamellar cell plasma membrane, and in the cisternae of the rough endoplasmic reticulum as well as in the nuclear envelope of lamellar cell bodies. No definite reaction products were detected within the axon terminals. These findings show that the lamellated corpuscles in the rat lip, like other mechanoreceptors, have an intense ChE activity which is mainly associated with lamellar cells. It can be said that ChE histochemistry is useful to detect mechanoreceptors. The functional significance of ChE in mechanoreceptors is discussed.

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Three-dimensional architecture of the endoneurium with special reference to the collagen fibril arrangement in relation to nerve fibers.

The endoneurium of the mouse sciatic nerve was studied by scanning and transmission electron microscopy to elucidate their three-dimensional architecture. The endoneurium consisted of collagen fibrils, and occasional fibroblasts and blood vessels. Collagen fibrils surrounded individual nerve fibers, forming two distinct layers of connective tissue sheath: the outer one was composed of bundles of longitudinally oriented collagen fibrils and the inner one was of a delicate network of interwoven thin collagen fibrils. These outer and inner layers of collagen fibrils correspond to the two fibrous sheaths known as the sheath of Key and Retzius and the sheath of Plenk and Laidlaw, respectively, revealed on nerve fibers by silver impregnation. Some of the finest collagen fibrils forming the inner network are closely attached to the basal lamina of Schwann cells, suggesting that these fibrils are concerned with the connection between the basal lamina and the inner collagen network. These two layers of collagen fibrils were found on all the nerve fibers, suggesting that they represent a general structure ensheathing the peripheral nerve fibers. Although these layers occur also on unmyelinated nerves, they are most developed on the largest myelinated fibers.

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Nerve regeneration through allogenic nerve grafts in mice.

The purpose of this study was to examine whether the basal laminae of Schwann cells in allografts could survive immunological rejection and serve as a conduit for regenerating nerves, as in the case of autogenic nerve grafts. Allografts of nerves were carried out using sciatic nerves of mice after the grafts had been repetitively frozen to kill their Schwann cells. Two mouse strains, C57BL/6N and C3H/HeN, were used, as they are known to differ in major histocompatibility complex. The mid-portion of the grafted nerve segments was examined by electron microscopy. In addition, the toe pad skin and lumbrical muscles were examined for determining whether regenerating nerves reinnervate sensory end organs and motor endplates. The process of nerve regeneration in the allograft was the same as that seen in the autograft. Cells in the graft disintegrated into cell debris and were phagocytized by macrophages, whereas the basal laminae of Schwann cells were not removed by macrophages, remaining in the form of tubes or scaffolds. Regenerating nerve fibers grew out through such basal lamina scaffolds, keeping in contact with the inner surface. Digital sensory corpuscles and motor endplates of the operated side were well reinnervated. The results indicate that the basal laminae of Schwann cells of the allograft may survive and serve as a conduit for regenerating axons in the same way as in the case of an autograft.

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