The primary cilium of connective tissue cells: imaging by multiphoton microscopy

The Anatomical Record : Advances in Integrative Anatomy and Evolutionary Biology
Eve DonnellyCornelia E Farnum

Abstract

Although the role of the primary cilium as a sensory organelle in epithelial cells has been elucidated significantly over the past decade, the function of primary cilia in connective tissue cells has been studied less extensively. Primary cilia have been implicated as mechanotransducers in connective tissues, but the mechanisms by which the cells sense loads and convert them to biochemical signals for tissue formation and adaptation are poorly understood. Before hypotheses regarding the function of the primary cilium in connective tissue cells can be tested, methods for quantitation of incidence as well as three-dimensional visualization of primary cilia with respect to the extracellular matrix (ECM) are needed. The objective of this study was to develop a rapid method for visualizing primary cilia in their native ECM in a wide range of connective tissues. Whole-mount immunohistochemical and multiphoton microscopy techniques were developed to simultaneously image primary cilia, cell nuclei, and collagen and their relationships to each other in situ. Axonemes of primary cilia projecting into the ECM were successfully visualized in thick sections of growth plate cartilage, tendon, ligament, meniscus, intervertebral disc, and peri...Continue Reading

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