• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제36권5호
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• pp.360-365
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• 2010

Introduction: Mammalian tooth pulp is densely innervated by sensory nerves that are mostly C fibers and A delta fibers. However, there is evidence suggesting that many unmyelinated axons in the pulp are in fact parent meylinated axons. Immunohistochemical staining for neurofilament protein 200 kDa (NFP200) was performed to identify the demyelinated but parent myelinated axons. Materials and Methods: The pulp was removed from healthy premolars and 3rd molars extracted from juveniles and adults undergoing orthodontic treatment, and immunohistochemical staining were applied with NPF200 antibodies, which specifically dye myelinated axons. The specimens underwent an electron microscopy examination with diaminobenzidine (DAB) immunostaining after observation and analysis by fluorescence and confocal laser scanning microscopy. Results: The NPF200 immuno-positive axons in the radicular pulp areas were observed as bundles of many nerve fibers. Many small bundles were formed with fewer axons when firing to the coronal pulp areas and then reachrd a different direction. In the radicular pulp, unmyelinated axons and myelinated axons were present together. However, in the coronal pulp, unmyelinated axons were most common and NFP200 immuno-positive unmyelinated axons with a larger diameter than those in the radicular pulp were observed more frequently. On the other hand, most of the immuno-positive unmyelinated fibers were similar in size to that of typically well-known unmyelinated fibers. Conclusion: Myelinated fibers innervated to the dental pulp maintain their myelins in the radicular portion, but these fibers lost myelins in the coronal portion. After the loss of myelin, the size of the axoplasm also decreased.

• Anatomy and Cell Biology
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• 제56권2호
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• pp.271-275
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• 2023

The abducens nerve (AN; cranial nerve VI) exits the brainstem at the inferior pontine sulcus, pierces the dura of the posterior cranial fossa, passes through the cavernous sinus in close contact to the internal carotid artery (ICA) and traverses the superior orbital fissure to reach the orbit to innervate the lateral rectus muscle. At its exit from the brainstem, the AN includes only axons from lower motor neurons in the abducens nucleus. However, as the AN crosses the ICA it receives a number of branches from the internal carotid sympathetic plexus. The arrangement, neurochemical profile and function of these sympathetic axons running along the AN remain unresolved. Herein, we use gross dissection and microscopic study of hematoxylin and eosin-stained sections and sections with tyrosine hydroxylase immunolabeling. Our results suggest the AN receives multiple bundles of unmyelinated axons that use norepinephrine as a neurotransmitter consistent with postganglionic sympathetic axons.

• Applied Microscopy
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• 제25권1호
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• pp.30-47
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• 1995

The ultrastructural changes of adrenal chromaffin cells and ganglion cells in the adrenal gland of vacor-induced diabetic Mongolian gerbils were studied by electron microscopy. After one month of vacor-induced diabetes, some chromaffin cells were filled with dense bodies and large cytosomes with formy contents. Most of degenerating axon terminals were observed on chromaffin cells. A few macrophages were found among chromaffin cells at one month after induction. Several of these macrophages were filled with numerous phagosomes. After one month of vacor-induced diabetes, the ganglion cells showed increase in numbers of dense bodies and degenerating dendrites compared with the normal ganglion cells. Both electron dense and lucent types of degenerating axon terminals were found in interstitial space of the ganglion cells. Degenerating unmyelinated and myelinted axons contained dense and lamellar bodies. The satellite cells and macrophages with engulfed degenerated axon terminals were observed. After three months of vacor-induced diabetes, the unmyelinated and myelinated axons showed degenerative changes, whereas no structural changes could be demonstrated in adrenal ganglion and chromaffin cells. The satellite cells and macrophages containing partially digested debris were still commonly observed in the interstitial space of adrenal medulla. These results suggest that the degenerative changes occur in the adrenal ganglion cells as well as adrenal chromaffin cells of vacor-induced diabetic Mongolian gerbils.

• Applied Microscopy
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• 제23권2호
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• pp.107-123
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• 1993

The ultrastructural changes of the cardiac ganglion and granule-containing cells in the heart of vacor-induced diabetic Mongolian gerbils were studied by electron microscopy. After one month of vacor-induced diabetes the ganglion cells showed increase in numbers of dense bodies and mitochondria compared with the normal cardiac ganglion. Most of the satellite cells were filled with numerous phagosomes containing digested debris. Both electron-dense and lucent types of degenerating axon terminals were observed. The former was characterized by clusters of agranular vesicles and numerous mitochondria. The electron lucent type of degenerating axon terminal contained a few agranular vesicles and swollen mitochondria. Degenerating unmyelinated and myelinated axons contained large numbers of dense bodies, lamellar bodies, and mitochondria. Numerous macrophages containing phagosomes were reveled in the interstitial spaces. Some of the granule-containing cells in the heart showed a variety of degenerative changes and a decreased number of dense-cored vesicles. After three months of vacor-induced diabetes the unmyelinated and myelinated axons showed degenerative changes, whereas no structure changes could be demonstrated in intraatrial ganglion and granule containing cells. The satellite cells containing engulfed debris were observed in the cardiac ganglion cells. These results suggest that the degenerative changes occur in the cardiac ganglion cells of vacor-induced diabetic Mongolian gerbils as well as atrial granule-containing cells.

• International Journal of Oral Biology
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• 제43권3호
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• pp.155-160
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• 2018

There exists very little information on the ultrastructure of substance P immunopositive (+) fibers in the human dental pulp, which may help in understanding the mechanism for substance P associated pulpal inflammatory pain. To address this issue, we investigated the presence of substance P+ fibers in the human dental pulp by light- and electron-microscopic immunohistochemistry. Light microscopy revealed that substance P+ fibers ran within neurovascular bundles in the radicular pulp and in the core of coronal pulp. They were also frequently present in the peripheral pulp. Substance P+ fibers showed beads like swellings interconnected by thin axonal strand, in a manner similar to bouton en passants and interconnecting axonal strand in the spinal cord. Electron microscopy revealed that almost all the substance P+ axons were unmyelinated. The axonal swellings of the substance P+ contained numerous clear round vesicles (40-50 nm in diameter) and many large dense-cored vesicles (80-110 nm in diameter) as well as many mitochondria. The vesicles and mitochondria were rarely observed in the thin axonal strand interconnecting the swellings. Intimate interrelationship or synaptic structure between the swellings of substance P+ axon and nearby pulpal cells or axons was not found. These findings suggest co-release of substance P and glutamate from the substance P+ pulpal axons and its action on nearby structures in a paracrine manner.

• Applied Microscopy
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• 제8권1호
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• pp.53-66
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• 1978

An attempt has been made to discriminate the synapses in the striatum consisting caudate nucleus, putamen and fundus striati of the cat with emphasis on the characteristic structures of axon terminals and postsynaptic profiles. The differentiation is based on the size and shape of vesicle in the bouton terminal, and the symmetrical or asymmetrical thickening the pre- and postsynaptic membrane. Four types of synapses could be differentiated: Type I: the bontons with asymmetrical,synaptic thickenings contain round 45 nm diameter vesicles and contact cell soma, dendritic shafts and dendritic spines (74%). Type II : the boutons contain round 45nm diameter vesicles and are associated with symmetrical membrane thickenings. These synapses are formed on the soma and dendritic shafts (6%). Type III: the boutons with symmetrical membrane thickenings contain 50-60 nm diameter pleomorphic vesicles, and contact soma and dendritic shafts (18%). Type IV: the terminals contain flattened vesicles ($25{\times}45 nm$) and are associated with symmetrical membrane thickenings. These synapses are found in contact with soma and dendritic shafts. Additionally, the bouton en passant, which is expanded from myelinated or unmyelinated axons containing round vesicles (45nm diameter) contacts the dendritic shaft or dendritic spine with asymmetrical membrane thickenings. Two unusual types of synapses, axo-axonic and dendro-dendritic, are found occasionally.

• Archives of Reconstructive Microsurgery
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• 제6권1호
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• pp.29-38
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• 1997

A peripheral nerve when approximation of the ends imparts tension at the anastomosis and with a relatively long segment defect after excision of neuroma and neurofibroma cannnot be repaired by early primary suture. The one of the optimistic reconstruction method of severed peripheral nerves is to restore tension-free continuity at the repair site putting an autogenous nerve graft into the neural gap despite of ancipating motor or sensory deficit of the donor nerve area. To overcome the deficit of the autogenous nerve graft, several other conduits supplying a metabolically active environment which is able to support axon regeneration and progression, providing protection against scar invasion, and guiding the regrowing axons to the distal stump of the nerve have been studied. An author have used ipsilateral femoral vein, ipsilateral femoral vein filled with fresh thigh muscle, and autogenous sciatic nerve for the sciatic nerve defect of around 10 mm in length to observe the regeneration pattern in rat by light and electron microscopy. The results were as follows. 1. Light microscopically regeneration pattern of nerve fibers in the autogenous graft group was more abundant than vein graft and vein filled with muscle group. 2. On ultrastructural findings, the proxial end of the graft in various groups showed similar regenerating features of the axons, myelin sheaths, and Schwann cells. The fascicular arrangement of the myelinated and unmyelinated fibers was same regardless of the type of conduits. There were more or less increasing tendency in the number and the diameter of myelinated fibers correlated with the regeneration time. 3. In the middle of the graft, myelinated nerve fibers of vein filled with muscle group were more in number and myelin sheath was thinner than in the venous graft, but the number of regenerating axons in autogenous nerve graft was superior to that in both groups of the graft. The amount of collagen fibrils and amorphous materials in the endoneurial space was increased to elapsed time. 4. There was no difference in regenerating patterns of the nerve fibers of distal end of the graft. The size and shape of the myelinated nerve fbers were more different than that of proximal and middle portion of the graft. From the above results, the degree of myelination and regenerating activity in autogenous nerve is more effective and active in other types of the graft and there were no morphological differences in either ends of the graft regardless of regeneration time.

• Applied Microscopy
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• 제25권1호
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• pp.1-14
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• 1995

The purpose of this study was to investigate the main sensory trigeminal nucleus in the aging rat brain by means of electron microscope. Male Sprague-Dawley rats, two (control group) and thirty six (aging group) months of age, were used. These animals were sacrificed by perfusion fixation with 2.5% glutaraldehyde-2.0% paraformaldehyde (0.1M phosphate buffer, pH 7.4) under sodium pentobarbital. The objective area was punched out with a sharp-edged metal cylinder of 0.8 mm in diameter. These blocks of tissue were then washed in 0.1M phosphate buffer, postfixed in 2% osmium tetroxide, dehydrated in a graded series of ethyl alcohol, and embedded in Epon 812. Thin sections were cut with Super Nova ultramicrotome, pick up on grids and double stained with lead citrate and uranyl acetate, and observed in JEOL 100B electron microscope. The results were as follows: 1. In the control group, the neuronal cell body of the main sensory trigeminal nucleus was filled with nucleus, Golgi complex, Nissl substance, mitochondria, microfilaments and microtubules. However, few Nissl substances are seen in neuronal cell body. Axoaxonic synapse, axodendritic synapse, axosomatic synapse, axospinous synapse, myelinated and unmyelinated nerve fibers were well organized around cell bodies. Neurons with abnormal changes were not seen. 2. In the aging group, the neuronal cell body of the main sensory trigeminal nucleus contained large number of lipofuscin granules, dense body and swollen mitochondria. Terminal boutons contained glycogen, crystal-like vesicle and membranous indicating first signs of degeneration. The dendrites were found to be in synaptic contact with altered axon terminals. Frequently axons filled with dark axoplasn and splitted myelin sheath were noticed.

• The Korean Journal of Pain
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• 제5권2호
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• pp.213-220
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• 1992

Morphine, meperidine 및 pentazocine을 가토의 좌골신경에 주입한 후 마약제의 신경차단 유무와 약제 주입 후 4시간, 24시간 및 1주에 좌골 신경을 절취하여 신경조직학적 변화를 관찰하였다. 좌골신경에 약제를 주입한 후 신경자극에 의한 반응과 뒷다리 운동을 관찰한 결과, morphine군은 신경차단 효과가 없었고 meperidine군과 pentazocine군은 약제주입 5분 후부터 근육이완이 시작되어 10분 후부터 근육수축이 나타나지 않았으며 뒷다리에 마비증상은 약제주입 60분 후부터 부분적으로 회복되기 시작하여 90분 후에는 정상으로 회복되는 양상의 신경차단 효과가 있었다. 광학 현미경적 소견으로는 모두 4시간부터 1주까지의 표본에 특기할만한 변화가 없었으며, 전자 현미경적 소견에서 morphine군은 1주 후 소견에서 유수신경섬유와 무수신경섬유에 경미한 수포양을 보였다. Meperidine군은 4시간 후 소견으로 유수신경섬유의 축삭돌기에 경미한 수포양이 있었고 무수신경의 마이엘린화되는 소견이 있었으며, 24시간 후 유수신경섬유에 경미한 수포양이 있었고 무수신경섬유가 정상으로 되었으며 1주 후 특기할 만한 변화가 없었다. Pentazocine군은 약제주입 4시간 후 유수신경섬유에 경미한 수포양을 보였으며 24시간 후 유수신경 섬유와 무수신경섬유에 중등도의 수포양이 나타났으며 1주 후 경미한 수포양을 나타내었다. 주입된 약제중 morphine이 가장 수포양이 적었으며 pentazocine이 심한 변화를 나타내었고, 전단계 쥐 실험에서 나타났던 meperidine주입 1주 후의 심한 신경조직 손상은 본 실험에서 나타나지 않았다.

• Applied Microscopy
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• 제24권4호
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• pp.61-77
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• 1994

The calcium-binding proteins (CaBP), parvalbumin (PV) and calbindin-D 28K (calbindin) are particularly abundant and specific in their distribution, and present in different subsets of neurons in many brain regions. Although their physiological roles in the neurons have not been elucidated, they are valuable markers of neuronal subpopulations for anatomical and developmental studies. This study is designed to characterize dorsal lateral geniculate nucleus (dLGN) neurons and axon terminals in terms of differential expression of immunoreactivity (IR) for two well-known CaBPs, PV and calbindin. The experiments were carried out on 6 adult monkeys. Monkeys were perfused under deep Nembutal anesthesia with 2% paraformaldehyde and 0.2% glutaraldehyde in 0.1M phosphate buffer. After removal, the brains were postfixed for 6-8 hr in 2% paraformaldehyde at $4^{\circ}C$ and infiltrated with 30% sucrose at $4^{\circ}C$. Thereafter, they were frozen in dry ice. Serial sections of the thalamus, at $20{\mu}m$, were made in the frontal plane with a sliding microtome. The sections were stained for PV and calbindin with indirect immunocytochemical methods. For electron microscopy, after infiltration with 30% sucrose the blocks of thalamus were serially sectioned at $50{\mu}m$ with a Vibratome in the coronal plane and stained immediately by indirect ABC methods without Triton X-100 in incubation medium. Stained sections were postfixed in 0.2% osmium tetroxide, dehydrated and flat-embedded in Spurr resin. The block was then trimmed to contain only a selected lamina or interlaminar space. The dLGN proper showed strong PV IR in fibers in all laminae and interlaminar zones. Particularly dense staining was noted in layers 1 and 2 that contain many stained fibers from optic tract. Neuronal cell body stained with PV was concentrated only in the laminae. In these laminae staining was moderate in cell bodies of all large and medium-sized neurons, and was strong in cell bodies of some small neurons together with their processes. Calbindin IR was marked in the neuronal cell body and neuropil in the S layers and interlaminar zones whereas moderate in the neuropil throughout the nucleus. Regional difference in distribution of PV and calbindin IR cell is distinct; the former is only in the laminae and the latter in both the S layer and interlaminar space. The CaBP-IR elements were confined to about $10{\mu}m$ in depth of Vibratome section. The IR product for CaBP was mainly associated with synaptic vesicle, pre- and post-synaptic membrane, and outer mitochondrial membrane and along microtubule. PV-IR was noted in various neuronal elements such as neuronal soma, dendrite, RLP, F, PSD and some myelinated or unmyelinated axons, and was not seen in the RSD and glial cells. Only a few neuronal components in dLGN was IR for calbindin and its reaction product was less dense than that of PV, and scattered throughout cytoplasm of soma of some relay neurons, and was also persent in some dendrite, myelinated axons and RLP. The RSD, F, PSD and glial elements were always non-IR for calbindin. Calbindin labelled RLP were presynaptic to unlabeled dendrite or dendritic spine and PSD. Calbindin-labeled dendrite of various sizes were always postsynaptic to unlabeled RSD, RLP or F. From this study it is suggested that dLGN cells of different functional systems and their differential projection to the visual cortex can be distinguished by differential expression of PV and calbindin.