• 혜화의학회지
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• 제14권1호
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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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• 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.

• 생명과학회지
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• 제19권7호
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• pp.859-865
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• 2009

미세소관은 세포골격단백질의 중요한 구성 단백질로 축삭돌기 내에서는 세포막 방향으로 정렬되어 있다. Kinesin superfamily (KIFs)는 세포 내에서 미세소관을 따라 세포 내 소포들을 운반하는 분자 자동차 (molecular motor) 단백질이다. 본 연구에서 우리는 효모 two-hybrid system을 사용하여 KIF1A의 coiled-coil 영역과 결합하는 단백질로 미세소관 불안정화 요소인 SCG10 단백질을 분리하였다. SCG10은 KIFs에서 KIF1A와만 특이적으로 결한 하며, KIF1A의 400에서 820아미노산 부위가 SCG10과의 결합에 필수적임을 효모 two-hybrid assay로 확인하였다. 또한 SCG10의 coiled-coil영역은 KIF1A와의 결합에 필수영역임을 확인하였으며 단백질간의 결합은 Glutathione S-transferase pull-down assay를 통하여 확인하였다. 생쥐의 뇌 파쇄액에 SCG10항체로 면역침강을 행하여 KIF1A를 확인한 결과KIF1A는 SCG10과 특이적으로 같이 침강하였다. 이러한 결과들은 KIF1A는 SCG10와 결합하여 SCG10이 포함된 소포를 미세소관을 따라 이동시킴을 시사한다.

• Molecules and Cells
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• 제38권10호
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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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• 제26권1호
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• pp.17-31
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• 1996

검은과부거미의 독 분비장치는 두흉부에 있는 가위턱과 한쌍의 독선으로 이루어져 있다. 독선은 한겹의 얇은 장막과 횡문근 섬유의 다발에 의해 둘러싸여 있었고. 근육층을 따라서 운동신경의 축삭 돌기와 근섬유 사이에 신경근육간 연접이 형성되어 있었다. 분비상피를 이루는 단층 원주상피세포에는 복잡한 수지상의 돌기가 형성되었고, 독선 전체가 단포상선을 이루고 있음이 관찰되었다. 상피의 분비면은 기저막으로부터 선의 내강쪽으로 확장된 세포질 돌기에 의해 표면적이 현저히 증가되었고, 상피의 내강면에는 조밀한 미세융모가 형성되어 있었다. 독성 분비물은 상피세포 내에서 두 종류의 분비과립으로부터 생성되었다. 분비기동안 이들 분비과립은 과적의 형태로 변형, 농축된 후, 이출분비의 형태로 내강으로 방출되었다. 방출 후의 기저부 상피세포들은 고도의 증식과정을 거쳐 원주상의 상피세포로 재생됨이 확인되었다.

• Applied Microscopy
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• 제29권2호
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• pp.223-230
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• 1999

본 연구는 경기도 양평에서 채집한 빨간집모기(Culex pipiens pallens: Culicidae)를 사육하여 유충 시기에 있는 직장 조직을 구성하는 세포들의 미세구조를 관찰하였다. 직장조직은 점막상피조직, 결합조직, 그리고 근육조직 등으로 구성되었으며, 상피 내면에는 않은 큐티클 내막이 덮고 있었다. 점막 상피조직은 단층의 편평상피세포들로 이루어졌으며, 첨단원형질막이 약 $5\sim8{\mu}m$ 길이의 막주름이 잡혀 있었고 그 사이에는 크게 발달한 사립체가 관찰되었다. 측부원형질막의 첨단 쪽에서는 격막연접부위가 길게 관찰되었으며 중간부위는 심하게 주름이 접혀 물질의 수송통로인 세포간극이 불규칙하게 관찰되었고 이 주변에서도 크게 발달한 사립체가 산재해 있었다. 그리고 상피세포의 기저막주름은 세포 내로 짧고 불규칙하게 함입되어 미로 형태로 관찰되었다. 기점막 바깥쪽에는 신경 및 기관소지가 분포하고 결합조직과 전형적인 골격근의 형태를 하고 있는 환상근관 종주근이 장외면에 부착되었다.

• 감성과학
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• 제7권2호
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• pp.179-186
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• 2004

중추신경계는 일단 손상이 되면 손상된 세포의 재생, 손상된 수초의 회복, 신경계의 정상적인 연결 등의 제한성 때문에 그 회복이 매우 힘들다. 이러한 중추신경계의 중요한 손상으로는 다발성 경화증, 뇌졸중, 척수손상, 외상, 축삭의 탈수초화 등이 있다. 이전 연구들은 많은 발생빈도를 보이고 있는 척수손상에서 실질적인 척수의 기능적인 회복을 위해 손상된 척수신경의 재생과 축삭의 재수초화가 중요한 요인이라고 전하고 있다. 최근에는 이러한 척수손상에 대한 치료적 접근으로서 세포이식 기술이 하나의 해결책을 열어주고 있다. 따라서 본 논문에서는 척수손상의 특성을 살펴보고, 척수손상에 의한 기능장애에 대해 세포이식이 기능의 회복을 증진시킬 수 있다는 증거를 논의하고자 한다.

• The Korean Journal of Physiology and Pharmacology
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• 제12권5호
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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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• 제24권6호
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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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• 제32권5호
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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.