• Molecules and Cells
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• v.40 no.6
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• pp.426-433
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• 2017

Ephrin-A5 has been implicated in the regulation of brain morphogenesis and axon pathfinding. In this study, we used bacterial homologous recombination to express a LacZ reporter in various ephrin-A5 BAC clones to identify elements that regulate ephrin-A5 gene expression during mesencephalon development. We found that there is mesencephalon-specific enhancer activity localized to a specific +25.0 kb to +30.5 kb genomic region in the first intron of ephrin-A5. Further comparative genomic analysis indicated that two evolutionary conserved regions, ECR1 and ECR2, were present within this 5.5 kb region. Deletion of ECR1 from the enhancer resulted in disrupted mesencephalon-specific enhancer activity in transgenic embryos. We also found a consensus binding site for basic helix-loop-helix (bHLH) transcription factors (TFs) in a highly conserved region at the 3'-end of ECR1. We further demonstrated that specific deletion of the bHLH TF binding site abrogated the mesencephalon-specific enhancer activity in transgenic embryos. Finally, both electrophoretic mobility shift assay and luciferase-based transactivation assay revealed that the transcription factor Ascl1 bound the bHLH consensus binding site in the mesencephalon-specific ephrin-A5 enhancer in vitro. Together, these results suggest that the bHLH TF binding site in ECR1 is involved in the positive regulation of ephrin-A5 gene expression during the development of the mesencephalon.

• Applied Microscopy
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• v.34 no.3
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• pp.199-209
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• 2004

The migut epitheluim of the last instar larva in the mosquito larvae, Anopheles sinensis was observed with electron microscopes. The midgut epitheluim of the mosquito larva is composed of a single-layered columnar absorptive cells, regenerative cells and secretory granular cells. The free surface of the columnar absorptive cells has a regular array of microvilli 'brush border', while cell membranes close to the basal lamina are extrmely infolded and a lot of mitochondria are concentrated in those processes. The columnar absorptive cells also contain cell organelles expected to be found in absorptive cell. Midgut regenerative cells which are positioned basally in the epithelium form the groups, which are called 'nidi', composed of 1 or $2{\sim}3$ cells, they show darker appearance than the columnar absoptive cells. The secretory granular cells contain numerous electron dense granules, $200{\sim}400$ nm in diameter. The cone shaped secretory granular cells are located in the basal portion of the midgut epitheluim. The epithelium is surrounded by the subepithelial space and muscle bundles. The subepithelial space, which is filled with fibrous connective tissue, is innervated by many axon cells and tracheoles.

• Archives of Plastic Surgery
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• v.33 no.2
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• pp.213-218
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• 2006

As the large defect of peripheral nerve occurs, the autologous nerve graft is the most ideal method but it has many limitations due to donor site morbidities. Various materials have been developed for the nerve defect as the conduits, but none of these materials is satisfactory. Among them, $Gore-Tex^{(R)}$ tube seems to be one of the most ideal nerve conduit materials at peripheral nerve defect. Many researches have focused on finding the neurotrophic factors. It is recently demonstrated that Valproic acid(VPA) has an effect of axonal regeneration as a neurotrophic factor without enzymatic degradation and toxicity problems. The purpose of this study is to evaluate the effect of VPA on the nerve regeneration at the peripheral nerve defect. A 10 mm gap of rat sciatic nerve was made and $Gore-Tex^{(R)}$ tube filled with biceps femoris muscle was placed at the nerve defect site. We let the rat take VPA as drinking water in experimental group and did not give VPA to the control group. We estimated the results as electrophysiologic and histological aspects for 16 weeks after the surgery. The nerve conduction velocity, total myelinated axon count, myelin sheath thickness and mean nerve fiber diameter significantly increased in VPA-treated experimental group when compared to the control (p < 0.05). From the above results, we conclude that VPA promotes the nerve regeneration at the peripheral nerve defect site. It is suggested that $Gore-Tex^{(R)}$ tube filled with skeletal muscle and VPA administration may be a good substitute for autologous nerve graft.

• Applied Microscopy
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• v.35 no.3
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• pp.177-186
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• 2005

The foregut epithelium of the last instar larva in the scarab beetle, Allomyrina dichotoma was observed with electron microscopes. The foregut epithelium of the scarab beetle larva is composed of a single-layered squamous absorptive cell. The luminal surface of the epithelium is covered with cuticular intima. The free surface of the squamous cell has a irregular array of microvilli 'brush border', while cell membrans close to the basal lamina are infolded and a lot of mitochondria are concentrated in those processes. The cytoplasm in the epithelial cells is well developed nucleus, mitochondria. And the basal region of cell contains large lipid-, protein droplets and numerous glycogen granules. The basal lamina is located between the basal membrane and muscle bundle, providing barrier between the epithelium and the hemolymph. The epithelium is surround by the subepithelial space and muscles. The subepithelial space, which is composed of fibrous connective tissue is innervated by many tracheoles and axon.

• Applied Microscopy
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• v.25 no.1
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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.

• Applied Microscopy
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• v.22 no.2
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• pp.30-45
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• 1992

An experimental study on the acute irradiation effects on the substantia nigra of head-irradiated rats were carried out. Rats anesthetized with sodium thiopental, were exposed only on their head areas with a single dose of 3,000 rads or 6,000 rads, respectively. Radiation was produced by Mitsubishi linear accelerator at the speed of 200 rads/min. Aminals were sacrificed on 6 hours, 2 days and 6 days following irradiations. By the perfusion fixation through the heart, rats were fixed with 1% glutaraldehyde-1% paraformaldehyde solution. Two hours later, brains were exposed and immersed in the same fixatives over night. Tissue blocks from subtantia nigra were punched out, and they were refixed in the 2% osmium tetroxide solution. Blocks were dehydrated through alcohol series, and embedded in the araldite mixture. Ultrathin sections were stained with uranyl acetate and lead citrate solutions, From the ultrastructural study, following results were made: 1. Six hours after irradiation, severe depletion of synaptic vesicles was occurred in the many axon terminals of the nigral neuropil. 2. Dramatical decrease of lysosomes and dense granules was observed. 3. Two days following irradiation, alterations of ribosomes, granular endoplasmic reticula, mitochondria, etc, were noticed. 4. Many of the malformations were seen to be repaired on the 6th day. 5. Above results were interpreted as follows. At the acute stage of heavy irradiation, neurotransmitters in the substantia nigra are released severely. But they are recovered within 6 days. It is concluded that acute head-irradiation may result severe disturbance of nigral motor control function during the first few days.

• Molecules and Cells
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• v.38 no.11
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• pp.1013-1021
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• 2015

Most of the axons in the vertebrate nervous system are surrounded by a lipid-rich membrane called myelin, which promotes rapid conduction of nerve impulses and protects the axon from being damaged. Multiple sclerosis (MS) is a chronic demyelinating disease of the CNS characterized by infiltration of immune cells and progressive damage to myelin and axons. One potential way to treat MS is to enhance the endogenous remyelination process, but at present there are no available treatments to promote remyelination in patients with demyelinating diseases. Sulfasalazine is an anti-inflammatory and immune-modulating drug that is used in rheumatology and inflammatory bowel disease. Its anti-inflammatory and immunomodulatory properties prompted us to test the ability of sulfasalazine to promote remyelination. In this study, we found that sulfasalazine promotes remyelination in the CNS of a transgenic zebrafish model of NTR/MTZ-induced demyelination. We also found that sulfasalazine treatment reduced the number of macrophages/microglia in the CNS of demyelinated zebrafish larvae, suggesting that the acceleration of remyelination is mediated by the immunomodulatory function of sulfasalazine. Our data suggest that temporal modulation of the immune response by sulfasalazine can be used to overcome MS by enhancing myelin repair and remyelination in the CNS.

• Molecules and Cells
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• v.37 no.6
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• pp.497-502
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• 2014

Microfluidics can provide unique experimental tools to visualize the development of neural structures within a microscale device, which is followed by guidance of neurite growth in the axonal isolation compartment. We utilized microfluidics technology to monitor the differentiation and migration of neural cells derived from human embryonic stem cells (hESCs). We co-cultured hESCs with PA6 stromal cells, and isolated neural rosette-like structures, which subsequently formed neurospheres in suspension culture. Tuj1-positive neural cells, but not nestin-positive neural precursor cells (NPCs), were able to enter the microfluidics grooves (microchannels), suggesting that neural cell-migratory capacity was dependent upon neuronal differentiation stage. We also showed that bundles of axons formed and extended into the microchannels. Taken together, these results demonstrated that microfluidics technology can provide useful tools to study neurite outgrowth and axon guidance of neural cells, which are derived from human embryonic stem cells.

• Clinical and Experimental Pediatrics
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• v.55 no.10
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• pp.397-402
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• 2012

Pelizaeus-Merzbacher disease (PMD) is a rare, X-linked recessive disorder characterized by dysmyelination in the central nervous system. PMD results from deletion, mutation, or duplication of the proteolipid protein gene (PLP1) located at Xq22, leading to the failure of axon myelination by oligodendrocytes in the central nervous system. PMD may be suspected when there are clinical manifestations such as nystagmus, developmental delays, and spasticity, and genetic analysis can confirm the diagnosis. Further diagnostic manifestations of the disease include a lack of myelination on brain magnetic resonance (MR) imaging and aberrant N-acetyl aspartate (NAA) and choline concentrations that reflect axonal and myelination abnormalities on phroton MR spectroscopy. We report 5 cases of PMD (in 1 girl and 4 boys). PLP1 duplication was detected in 2 patients. Brain MR analyses and MR spectroscopy were performed for all the patients. The brain MR images showed white matter abnormalities typical of PMD, and the MR spectroscopic images showed diverse patterns of NAA, creatinine, and choline concentrations. We propose that MR spectroscopic analysis of metabolic alterations can aid the PMD diagnosis and can contribute to a better understanding of the pathogenesis of the disease.

• Journal of Medicine and Life Science
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• v.16 no.3
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• pp.96-100
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• 2019

Postural tachycardia syndrome (POTS) is common, although not so well-known variant of cardiovascular autonomic disorder characterized by an excessive heart rate increase on standing. POTS is probably underdiagnosed due to the heterogeneity in both presentation and etiology. This study aimed to evaluate the clinical and autonomic features in patients with POTS. We reviewed the medical records of patients with POTS. Medical records include onset age, sex, presenting symptoms, body mass index (BMI) and prognosis. All patients had an autonomic function and laboratory tests. Ninety-nine patients met the inclusion criteria for POTS (51.5% male; mean±SD age, 20.0±9.7 years; mean±SD, BMI 21.9±3.9). Common presenting symptoms were a brief loss of consciousness, dizziness, blurred vision and headache. Autonomic function tests showed abnormal quantitative sudomotor axon reflex testing in 20 patients of 99 POTS patients. The abnormal post-ganglionic sympathetic sudomotor function is generally considered to reflect a neuropathic form of POTS. In treatments, 83 patients were treated by non-pharmacological management including lifestyle changes and 16 patients required the initiation of pharmacological therapies. Most patients with POTS showed a relatively favorable prognosis. POTS is a chronic disease with a substantial subset of patients recovering within a few years after the initial presentation. Future efforts should focus on better understanding of POTS pathophysiology and designing randomized controlled trials for the selection of more effective therapy.