CAJ | 학술논문

目的：探讨轴突对施万细胞(Schwann cells, SC)分化成熟与髓鞘形成的影响，及在SC的极向分化过程中整合素α6β4的表达。方法：用Wistar乳鼠坐骨神经分离SC，用其脊髓分离神经细胞。经纯化后分别培养及共同培养。用扫描电镜观察二者的形态及髓鞘形成情况；分别用髓鞘碱性蛋白(myelin basic protein,MBP)、神经微丝(neurofilament,NF）抗体进行免疫标记；采用原位杂交技术检测整合素α6β4 mRNA的表达。结果：单独培养的SC MBP抗原表达阴性；原位杂交检测α6β4表达阴性。SC与神经细胞共同培养时其MBP阳性（说明其已转变为髓鞘形成细胞），在神经突起周围表达阳性；扫描电镜可见SC与神经细胞贴附、融合、包绕轴突形成髓鞘；原位杂交显示α6β4在SC与神经轴突相贴附处呈线状阳性表达。结论：SC的分化成熟及髓鞘形成呈轴突依赖性，仅在与神经细胞共同培养时，才开始形成髓鞘。它在包绕轴突形成髓鞘时，α6β4在SC表面沿基膜呈方向性表达，在SC与基膜、细胞外基质之间起信号传导作用，使增生的SC具有极性并发生形态结构的改变。在SC的分化成熟及髓鞘形成的过程中，轴突及细胞外基质的作用不可忽视。
목적：탐토축돌대시만세포(Schwann cells, SC)분화성숙여수초형성적영향，급재SC적겁향분화과정중정합소α6β4적표체。방법：용Wistar유서좌골신경분리SC，용기척수분리신경세포。경순화후분별배양급공동배양。용소묘전경관찰이자적형태급수초형성정황；분별용수초감성단백(myelin basic protein,MBP)、신경미사(neurofilament,NF）항체진행면역표기；채용원위잡교기술검측정합소α6β4 mRNA적표체。결과：단독배양적SC MBP항원표체음성；원위잡교검측α6β4표체음성。SC여신경세포공동배양시기MBP양성（설명기이전변위수초형성세포），재신경돌기주위표체양성；소묘전경가견SC여신경세포첩부、융합、포요축돌형성수초；원위잡교현시α6β4재SC여신경축돌상첩부처정선상양성표체。결론：SC적분화성숙급수초형성정축돌의뢰성，부재여신경세포공동배양시，재개시형성수초。타재포요축돌형성수초시，α6β4재SC표면연기막정방향성표체，재SC여기막、세포외기질지간기신호전도작용，사증생적SC구유겁성병발생형태결구적개변。재SC적분화성숙급수초형성적과정중，축돌급세포외기질적작용불가홀시。
Objective: To study the axonal effect and the expression of integrin α6β4 during Schwann cell(SC) differentiation and myelination. Methods: Schwann cells were dissociated from the sciatic nerve of neonatal Waster rats and neurons dissociated from spinal cord. Singal cultures and purified populations of SC were cocultured with NC. Four methods (contrast microscope, scanning electron microscopy(SEM), immunocytochemistry method and in situ hybridization ) were used. Results: The separately cultured Schwann cells showed MBP negetive by immunocytochemistry method. But cocultured SC were shown positive. SEM showed that Schwann cells' membrane loop progressively circumnavigated around the axon during myelination, which suggested that the non-myelinating SC(nMSC) transformed to myelinating SC (MSC). In situ hybridization showed integrin α6β4 positive signals only on the outer surface of the Schwann cell-axon unit in SC coculture with NC. Conclusion: The differentiation and maturation of SC depend on axon, and the activity of integrins is expressed by axon. Axonal contact induces the expression of α6β4 during SC myelination, which suggests that integrin α6β4 is an important mediator of interactions of myelinating SC with the basal limina.