Analysis of the orientation of primary cilia in growth plate cartilage: A mathematical method based on multiphoton microscopical images
Analysis of the orientation of primary cilia in growth plate cartilage: A mathematical method based on multiphoton microscopical images
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DOI：
10.1016/j.jsb.2006.11.004
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发表时间：
2007-06-01
影响因子：
3
通讯作者：
Farnum, Cornelia
中科院分区：
文献类型：
--
作者：
Ascenzi, Maria-Grazia;Lenox, Michelle;Farnum, Cornelia
软骨细胞的初级纤毛已被假设为作为一个机械传感器，类似于上皮组织中的细胞的初级纤毛。我们假设，在生长过程中的机械输入，通过初级纤毛，导致定向分泌的细胞外基质，从而建立生长板软骨的组织各向异性。纤毛，通过其在三维空间中的方向，被假设为传递给软骨细胞的基质分泌的优先方向。本文报道了经典的数学方法的应用，开发一种算法，解决了特定的挑战相对于评估的主要纤毛在生长板软骨的方向，基于图像分析的光学切片可视化的多光子显微镜。通过基于快速冷沉淀的固定来制备标本，以最大限度地减少可能的睫状体方向的人为死后改变。纤毛轴丝通过免疫细胞化学与抗体乙酰化α-微管蛋白定位。该方法适用于调查纤毛方向在不同区域的生长板，无论是正常的或改变的生物力学环境。该方法是高度灵活的，适用于其他结缔组织，其中组织各向异性和细胞外基质成分的定向分泌被假设为取决于组织在发育和生长期间的生物力学环境。(c)2006年爱思唯尔公司All rights reserved.
The chondrocytic primary cilium has been hypothesized to act as a mechano-sensor, analogously to primary cilium of cells in epithelial tissues. We hypothesize that mechanical inputs during growth, sensed through the primary cilium, result in directed secretion of the extracellular matrix, thereby establishing tissue anisotropy in growth plate cartilage. The cilium, through its orientation in three-dimensional space, is hypothesized to transmit to the chondrocyte the preferential direction for matrix secretion. This paper reports on the application of classical mathematical methods to develop an algorithm that addresses the particular challenges relative to the assessment of the orientation of the primary cilium in growth plate cartilage, based on image analysis of optical sections visualized by multiphoton microscopy. Specimens are prepared by rapid cold precipitation-based fixation to minimize possible artifactual post-mortem alterations of ciliary orientation. The ciliary axoneme is localized by immunocytochemistry with antibody acetylated-alpha-tubulin. The method is applicable to investigation of ciliary orientation in different zones of the growth plate, under either normal or altered biomechanical environments. The methodology is highly flexible and adaptable to other connective tissues where tissue anisotropy and directed secretion of extracellular matrix components are hypothesized to depend on the tissue's biomechanical environment during development and growth. (c) 2006 Elsevier Inc. All rights reserved.