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首页> 外文期刊>Spine >Cell polarity in the anulus of the human intervertebral disc: morphologic, immunocytochemical, and molecular evidence.
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Cell polarity in the anulus of the human intervertebral disc: morphologic, immunocytochemical, and molecular evidence.

机译：人类椎间盘环的细胞极性：形态学，免疫细胞化学和分子证据。

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STUDY DESIGN: Human intervertebral disc tissue was obtained in a prospective study of cell morphology and gene expression. Experimental studies were approved by the authors' Human Subjects Institutional Review Board. Discs were obtained from surgical specimens or control donors. OBJECTIVES: To determine if there is morphologic and molecular evidence for polarity in cells of the human anulus. SUMMARY OF BACKGROUND DATA: In many tissues, cells become polarized as they develop functional specializations, which involve cell-cell and cell-extracellular matrix interactions and polarized targeting mechanisms. The highly specialized lamellar organization of the anulus is well recognized and suggests that this structure may be the result of directed secretion of extracellular matrix components by polarized disc cells. METHODS: Human disc specimens from donor and surgical patients were examined with light and electron microscopy to assess morphology. Specimens were examined for immunocytochemical localization ofPAR3 and claudin-1 and -11, recognized polarity proteins, and additional anulus specimens were examined for expression of polarity-related genes using microarray analysis. In vitro monolayer and 3-dimensional anulus cultures were also studied for gene expression, and additional surgical anulus specimens were used to obtain gene expression data using real time RT-PCR. RESULTS: At the ultrastructural level, anulus cells showed localization of secretory organelles and directed deposition of extracellular matrix in one portion of the cell, with the nucleus positioned in the opposite side of the cell. Expression of the polarity-related genes claudin-11 and PAR3 and PARD6 was confirmed using RT-PCR and microarray studies and immunocytochemical analyses. The percentage of cells positive for PAR3 immunolocalization was significantly greater in the outer anulus (100%) than in either the inner anulus (43.8%) or nucleus pulposus (22.6%). CONCLUSIONS: At the macroscopic level, the characteristic anular lamellar morphology implies a specialized architectural formation and organization, which is achieved by the tissue-specific function of polarized cells. Morphologic and molecular studies provided evidence for the presence of polarity in cells in the anulus. These findings advance our understanding of anulus disc cell function in production of highly aligned collagen fibrils and macroaggregates of these collagen fibrils into lamellar collagen bundles. Such disc cell activity is important in development and maintenance of the tissue-specific extracellular matrix of the disc.
机译：研究设计：通过对细胞形态和基因表达的前瞻性研究获得了人椎间盘组织。实验研究得到作者的人类受试者机构审查委员会的批准。光盘取自手术标本或对照供体。目的：确定在人类肛门环细胞中是否有形态学和分子学证据证明极性。背景数据概述：在许多组织中，细胞在发展功能专长时会变得极化，这涉及细胞-细胞和细胞-细胞外基质的相互作用以及极化的靶向机制。环的高度专业化的层状组织已得到公认，并表明该结构可能是极化的盘状细胞直接分泌细胞外基质成分的结果。方法：用光镜和电子显微镜检查供体和手术患者的人椎间盘标本，以评估形态。检查了样本的PAR3和claudin-1和-11的免疫细胞化学定位，识别了极性蛋白，并使用微阵列分析检查了其他无环标本的极性相关基因的表达。还研究了体外单层和3维肛门环培养物的基因表达，并使用实时RT-PCR使用其他外科肛门环标本获取基因表达数据。结果：在超微结构水平上，肛门环细胞显示出分泌细胞器的定位和细胞外基质在细胞一部分中的定向沉积，细胞核位于细胞的另一侧。使用RT-PCR和微阵列研究以及免疫细胞化学分析证实了极性相关基因claudin-11，PAR3和PARD6的表达。外环（100％）中PAR3免疫定位阳性的细胞百分比显着大于内环（43.8％）或髓核（22.6％）。结论：在宏观水平上，特征性的层状薄片形态暗示了专门的结构形成和组织，这是通过极化细胞的组织特异性功能来实现的。形态学和分子研究为环上细胞中极性的存在提供了证据。这些发现提高了我们对圆盘细胞在高度排列的胶原原纤维以及这些胶原原纤维的大聚集体形成层状胶原束的聚集中的功能的理解。这种椎间盘细胞活性在椎间盘的组织特异性细胞外基质的发育和维持中很重要。

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