Abstract：
OBJECTIVE: The goal of this study was to define basic constituents of the adult peripheral nervous system (PNS) using intact human nerve tissues.
METHODS: We combined fluorescent and chromogenic immunostaining methods, myelin-selective fluorophores, and routine histological stains to identify common cellular and noncellular elements in aldehyde-fixed nerve tissue sections. We employed Schwann cell (SC)-specific markers, such as S100β, NGFR, Sox10, and myelin protein zero (MPZ), together with axonal, extracellular matrix (collagen IV, laminin, fibronectin), and fibroblast markers to assess the SC\'s relationship to myelin sheaths, axons, other cell types, and the acellular environment.
RESULTS: Whereas S100β and Sox10 revealed mature SCs in the absence of other stains, discrimination between myelinating and non-myelinating (Remak) SCs required immunodetection of NGFR along with axonal and/or myelin markers. Surprisingly, our analysis of NGFR+ profiles uncovered the existence of at least 3 different novel populations of NGFR+/S100β- cells, herein referred to as nonglial cells, residing in the stroma and perivascular areas of all nerve compartments. An important proportion of the nerve\'s cellular content, including circa 30% of endoneurial cells, consisted of heterogenous S100β negative cells that were not associated with axons. Useful markers to identify the localization and diversity of nonglial cell types across different compartments were Thy1, CD34, SMA, and Glut1, a perineurial cell marker.
CONCLUSIONS: Our optimized methods revealed additional detailed information to update our understanding of the complexity and spatial orientation of PNS-resident cell types in humans.
METHODS: We combined fluorescent and chromogenic immunostaining methods, myelin-selective fluorophores, and routine histological stains to identify common cellular and noncellular elements in aldehyde-fixed nerve tissue sections. We employed Schwann cell (SC)-specific markers, such as S100β, NGFR, Sox10, and myelin protein zero (MPZ), together with axonal, extracellular matrix (collagen IV, laminin, fibronectin), and fibroblast markers to assess the SC\'s relationship to myelin sheaths, axons, other cell types, and the acellular environment.
RESULTS: Whereas S100β and Sox10 revealed mature SCs in the absence of other stains, discrimination between myelinating and non-myelinating (Remak) SCs required immunodetection of NGFR along with axonal and/or myelin markers. Surprisingly, our analysis of NGFR+ profiles uncovered the existence of at least 3 different novel populations of NGFR+/S100β- cells, herein referred to as nonglial cells, residing in the stroma and perivascular areas of all nerve compartments. An important proportion of the nerve\'s cellular content, including circa 30% of endoneurial cells, consisted of heterogenous S100β negative cells that were not associated with axons. Useful markers to identify the localization and diversity of nonglial cell types across different compartments were Thy1, CD34, SMA, and Glut1, a perineurial cell marker.
CONCLUSIONS: Our optimized methods revealed additional detailed information to update our understanding of the complexity and spatial orientation of PNS-resident cell types in humans.
摘要：
目的：本研究的目的是使用完整的人神经组织确定成人周围神经系统（PNS）的基本组成。
方法：我们结合了荧光和显色免疫染色方法，髓鞘选择性荧光团，和常规组织学染色，以鉴定醛固定的神经组织切片中常见的细胞和非细胞元素。我们采用了施万细胞（SC）特异性标记，如S100β，NGFR,Sox10和髓磷脂蛋白零（MPZ），连同轴突，细胞外基质(胶原蛋白IV，层粘连蛋白,纤连蛋白），和成纤维细胞标记以评估SC与髓鞘的关系，轴突,其他细胞类型，和无细胞环境。
结果：而S100β和Sox10在没有其他染色的情况下显示出成熟的SC，髓鞘化和非髓鞘化(Remak)SCs之间的区别需要对NGFR以及轴突和/或髓鞘标志物进行免疫检测.令人惊讶的是,我们对NGFR+概况的分析揭示了至少3种不同的NGFR+/S100β-细胞新群体的存在，本文中称为非神经胶质细胞，存在于所有神经区室的间质和血管周围区域。神经细胞含量的重要比例，包括大约30%的神经内膜细胞，由与轴突无关的异质S100β阴性细胞组成。鉴定不同区室非胶质细胞类型的定位和多样性的有用标记是Thy1，CD34，SMA，和Glut1，一种神经周细胞标记。
结论：我们优化的方法揭示了其他详细信息，以更新我们对人类PNS驻留细胞类型的复杂性和空间取向的理解。
方法：我们结合了荧光和显色免疫染色方法，髓鞘选择性荧光团，和常规组织学染色，以鉴定醛固定的神经组织切片中常见的细胞和非细胞元素。我们采用了施万细胞（SC）特异性标记，如S100β，NGFR,Sox10和髓磷脂蛋白零（MPZ），连同轴突，细胞外基质(胶原蛋白IV，层粘连蛋白,纤连蛋白），和成纤维细胞标记以评估SC与髓鞘的关系，轴突,其他细胞类型，和无细胞环境。
结果：而S100β和Sox10在没有其他染色的情况下显示出成熟的SC，髓鞘化和非髓鞘化(Remak)SCs之间的区别需要对NGFR以及轴突和/或髓鞘标志物进行免疫检测.令人惊讶的是,我们对NGFR+概况的分析揭示了至少3种不同的NGFR+/S100β-细胞新群体的存在，本文中称为非神经胶质细胞，存在于所有神经区室的间质和血管周围区域。神经细胞含量的重要比例，包括大约30%的神经内膜细胞，由与轴突无关的异质S100β阴性细胞组成。鉴定不同区室非胶质细胞类型的定位和多样性的有用标记是Thy1，CD34，SMA，和Glut1，一种神经周细胞标记。
结论：我们优化的方法揭示了其他详细信息，以更新我们对人类PNS驻留细胞类型的复杂性和空间取向的理解。
