• Journal of Haehwa Medicine
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• v.14 no.1
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• pp.201-211
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• 2005

Cdc2 kinase is a prototypical cyclin-dependent kinase critical for G2 to M phase cell cycle transition. Yet, its function in the nervous system is largely unknown. Here, we investigated possible role of Cdc2 in axonal regeneration using sciatic nerve system in rat. Cdc2 protein levels and activity were increased in the injured sciatic nerves 3 and 7 days after crush injury and then decreased to basal level 14 days later. Administration of Cdc2 kinase inhibitor roscovitine in vivo at the time of crush injury significantly inhibited axonal regeneration when regrowing axons were analyzed using retrograde tracers. Cdc2 protein levels in cultured Schwann cells which were prepared from sciatic nerves 7 days after crush injury were much higher compared with those from uninjured sciatic nerves, suggesting that Cdc2 protein expression was primarily induced in the Schwann cells. To further investigate Cdc2 function in Schwann cell, we examined changes in cultured Schwann cell proliferation and migration in culture system. Both the number of proliferating Schwann cells and the extent of neurite outgrowth from co-cultured DRG neurons were significantly decreased by Cdc2 inhibitor roscovitine treatment in DRG culture which was prepared from animals with sciatic nerve injury for 7 days. Also, Schwann cell migration in the injured sciatic nerve explant was significantly inhibited by roscovitine treatment. Taken together, the present data suggest that Cdc2 may be involved in peripheral nerve regeneration via Schwann cell proliferation and migration.

• International Journal of Oral Biology
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• v.43 no.3
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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.

• Journal of Life Science
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• v.19 no.7
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• pp.859-865
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• 2009

Microtubules, a major cytoskeleton, form parallel arrays in the axon and are oriented with their plus ends toward the cell periphery. Kinesin superfamily proteins (KIFs) are the molecular motors acting in the microtubule-based motilities of organelles in cells. Here, we used the yeast two-hybrid system to identify the protein that interacts with the coiled-coil domain of KIF1A and found a specific interaction with microtubule-destabilizing factor SCG10. SCG10 bound to the amino acid residues between 400 and 820 of KIF1A, but not to other KIFs in the yeast two-hybrid assay. The coiled-coil domain of SCG10 is essential for interaction with KIF1A. In addition, this specific interaction was also observed in the Glutathione S-transferase pull-down assay. An antibody to SCG10 specifically co-immunoprecipitated KIF1A associated with SCG10 from mouse brain extracts. These results suggest that KIF1A motor protein transports SCG10-containing vesicles along microtubules in neurons.

• Molecules and Cells
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• v.38 no.10
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• pp.876-885
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• 2015

N-acetyl-D-glucosamine kinase (NAGK) plays an enzyme activity-independent, non-canonical role in the dendritogenesis of hippocampal neurons in culture. In this study, we investigated its role in axonal development. We found NAGK was distributed throughout neurons until developmental stage 3 (axonal outgrowth), and that its axonal expression remarkably decreased during stage 4 (dendritic outgrowth) and became negligible in stage 5 (mature). Immunocytochemistry (ICC) showed colocalization of NAGK with tubulin in hippocampal neurons and with Golgi in somata, dendrites, and nascent axons. A proximity ligation assay (PLA) for NAGK and Golgi marker protein followed by ICC for tubulin or dynein light chain roadblock type 1 (DYNLRB1) in stage 3 neurons showed NAGK-Golgi complex colocalized with DYNLRB1 at the tips of microtubule (MT) fibers in axonal growth cones and in somatodendritic areas. PLAs for NAGK-dynein combined with tubulin or Golgi ICC showed similar signal patterns, indicating a three way interaction between NAGK, dynein, and Golgi in growing axons. In addition, overexpression of the NAGK gene and of kinase mutant NAGK genes increased axonal lengths, and knockdown of NAGK by small hairpin (sh) RNA reduced axonal lengths; suggesting a structural role for NAGK in axonal growth. Finally, transfection of 'DYNLRB1 (74-96)', a small peptide derived from DYNLRB1's C-terminal, which binds with NAGK, resulted in neurons with shorter axons in culture. The authors suggest a NAGK-dynein-Golgi tripartite interaction in growing axons is instrumental during early axonal development.

• Applied Microscopy
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• v.26 no.1
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• pp.17-31
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• 1996

The venomous apparatus of the black widow spider, Latrodectus mactans, is composed of chelicera and paired venom glands in the cephalothorax. Each glands is surrounded by a thin adventitia and striated muscular bundles resting on a basal membrane. Along the musculature neuromuscular synaptic contacts are formed by a motor axon and the muscle fibers. The secretory epithelium, which made up of simple and long columnar cells with extensive finger-like processes, creates a simple acinar gland. The secretory surfac is increased by a sort of fringes extended from the basal membrane into the gland lumen, and the luminal surface of the epithelium is marked by the presence of closely spaced microvilli. The venoms of the black widow spider are produced from two types of secretory granules within the epithelial cells. During the secretory phase, these granules are transformed into droplets and suffering a condensation. Finally the secretory products are released by the apocrine secretion. After the gland is emptied, the basal epithelial cells present a high proliferative process and regenerate the columnar epithlial cells.

• Applied Microscopy
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• v.29 no.2
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• pp.223-230
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• 1999

The epithelium of the rectum in the mosquito larvae, Culex pipiens pallens: Culicidae, was observed with electron microscope. The rectum of posterior hindgut was composed of epithelial tissue which were covered with cuticular intima on the luminal side, connective tissue and muscular tissue. The rectal epithelial cells were squamous absorptive cells, and apical plasma membranes were highly folded to form apical infoldings with mitochondria inserted them. The lateral plasma membranes were irregularly infolded and well developed mitochondria were found closely associated with infoldings . And intercellular spaces (or channels) were formed between the epithelial cells, whereas speptate junction was found near the apical zone between them. Also basal plasma membrane were infolded which made basal infoldings ('basal labyrinth'), and were covered with thin basal lamina. Rcetal epithelium was surrounded by the connective tissue which was contained axon and tracheole cells. Connective tissue was covered with the bundles of circular and longitudinal muscles.

• Science of Emotion and Sensibility
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• v.7 no.2
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• pp.179-186
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• 2004

Acute spinal cord injury can produce neurologic injury with many physical, psychological and social ramifications. It has been shown that two separate components combine to produce neurologic damage in acute spinal cord injury : the primary and secondary injuries. The primary mediators of spinal cord injury include the actual mechanical tissue disruption which is a passive process that occurs immediately following the trauma. A secondary injury cascade follows which appears mediated by cellular and molecular processes working through complex mechanisms. Both the primary and secondary injury cascades produce cell death both in neuronal and supporting cell tissues. Recovery from central nervous system(CNS) disorders is hindered by the limited ability of the vertebrate CNS to regenerate injured cells, replace damaged myelin sheath, and re-establish functional neuronal connections. Of many CNS disorders including multiple sclerosis, stroke, and other trauma, spinal cord injury is one of the important diseases because of the direct association with the functional loss of the body. Previous studies suggest that substantial recovery of function might be achieved through regeneration of lost neuronal cells and remyelination of intact axon in spinal cord injury which is occurred frequently. As a therapeutic approach in spinal cord injury, recently, cell transplantation provides a potential solution for the treatment of spinal cord injury. This review describes the characteristics of spinal cord injury and presents some evidence supporting functional recovery after cell transplantation following spinal cord injury.

• The Korean Journal of Physiology and Pharmacology
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• v.12 no.5
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• pp.225-230
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• 2008

Netrins are secreted molecules and involved in axon guidance, cell migration and tumor development. Recent studies revealed that netrins perform novel functions in such processes as epithelial development and angiogenesis without operating through the classical netrin receptors, DCC (Deleted in Colorectal Cancer) and Unc5h. In the present study, we investigated the roles of netrin-1 and its receptors in cell spreading of human glioblastoma cells, and found that netrin-1 haptotactically enhanced fibronectin-induced cell spreading and focal adhesion formation in U373 glioblastoma cells. Netrin-1 binding to the U373 cell membrane was blocked by an antibody against ${\alpha}v$ integrin subunit, but not by an anti-DCC or anti-Unc5h antibody. In addition, enhancement of the fibronectin response by netrin-1 was abrogated by a function blocking antibody against integrin ${\alpha}v{\beta}3$. Since the ${\alpha}v$ subunit of the integrin family plays an important role in the pathophysiological aspects of cell migration, including tumor angiogenesis and metastasis, our data provide important insight into the molecular mechanism of netrin function.

• The Korean Journal of Physiology and Pharmacology
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• v.24 no.6
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• pp.517-527
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• 2020

Layer 2/3 pyramidal neurons (L2/3 PyNs) of the cortex extend their basal dendrites near the soma and as apical dendritic tufts in layer 1, which mainly receive feedforward and feedback inputs, respectively. It is suggested that neuromodulators such as serotonin and acetylcholine may regulate the information flow between brain structures depending on the brain state. However, little is known about the dendritic compartment-specific induction of synaptic transmission in single PyNs. Here, we studied layer-specific serotonergic and cholinergic induction of long-term synaptic plasticity in L2/3 PyNs of the agranular insular cortex, a lateral component of the orbitofrontal cortex. Using FM1-43 dye unloading, we verified that local electrical stimulation to layers 1 (L1) and 3 (L3) activated axon terminals mostly located in L1 and perisomatic area (L2/3). Independent and AMPA receptor-mediated excitatory postsynaptic potential was evoked by local electrical stimulation of either L1 or L3. Application of serotonin (5-HT, 10 μM) induced activity-dependent longterm depression (LTD) in L2/3 but not in L1 inputs. LTD induced by 5-HT was blocked by the 5-HT₂ receptor antagonist ketanserin, an NMDA receptor antagonist and by intracellular Ca²⁺ chelation. The 5-HT₂ receptor agonist α-me-5-HT mimicked the LTD induced by 5-HT. However, the application of carbachol induced muscarinic receptor-dependent LTD in both inputs. The differential layer-specific induction of LTD by neuromodulators might play an important role in information processing mechanism of the prefrontal cortex.

• Archives of Plastic Surgery
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• v.32 no.5
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• pp.613-618
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• 2005

When a large peripheral nerve defect occurs, an autologous nerve graft is the most ideal method of recinstruction. But an autologous nerve graft has many limitations due to donor site morbidities. Many previous focused on finding the ideal nerve conduit. Among them, $Gore-Tex^{(R)}$ has several advantages over other conduits. It can be manipulated to a suitable size, does not collapse easily, and it is a semi- permeable material that contain pores. A round shaped nerve can be newly formed because of its smooth inner surface. The purpose of this study was to evaluate the availability of $Gore-Tex^{(R)}$ tube as a nerve conduit at the peripheral nerve defect in the rat sciatic nerve. The 10 mm nerve gap was made in each group. A $Gore-Tex^{(R)}$ tube filled with skeletal muscle was inserted and autologous nerve graft was harvested, respectively. In the experimental group, we placed a 0.5 mm thickness, $30{\mu}m$ pored, 1.8 mm in diameter and 14 mm length tube with skeletal muscle inserted inside. In the control group, the nerve gap was inserted with a rat sciatic nerve. We estimated the results electrophysiologically and histologically to 16 weeks postoperatively. Results in the nerve conduction velocity, total myelinated axon count, myelin sheath thickness and mean nerve fiber diameter, the experimental group was substantially lower than that of the control group, but the statistic difference was not significant (p<0.05). The morphology was very similar in both groups, microscopically. From the above results, We conclude that $Gore-Tex^{(R)}$ qualifies as an ideal nerve conduit. It is suggested that $Gore-Tex^{(R)}$ tube filled with skeletal muscle may, substitute for an autologous nerve graft.