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The cytoskeletal architecture can be studied using different microscopic and biochemical techniques. Electron microscopy was instrumental in discovering the cytoskeletal architecture around the 1960s, which allowed obtaining structural information at a high-resolution level. However, the sample preparation procedure often limits this ability in biological samples. Several protocols have been developed over the years to optimize sample preparation. In one of the protocols known as rotary...
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Complex microtubule structures are present in resting cells and in dividing cells. In resting cells, they are responsible for maintaining the cellular architecture, tracks for intracellular transport, positioning of organelles, assembly of cilia and flagella. They mediate the bipolar spindle assembly for chromosomal segregation and positioning of the cell division plate in dividing cells. The formation of microtubule complex structures depends on the cell type, cell stage, and cell function.
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Functionalization of Atomic Force Microscope Cantilevers with Single-T Cells or Single-Particle for Immunological Single-Cell Force Spectroscopy
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Tetrahymena Cilia Cap is Built in a Multi-step Process: A Study by Atomic Force Microscopy.

Cecília Seixas1, João Gonçalves2, Luís Viseu Melo3

  • 1Instituto Gulbenkian de Ciência, Apartado 14, 2781-901 Oeiras, Portugal.

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|November 18, 2017
PubMed
Summary

Cilia assembly involves transient structures forming at the distal tip before the central pair microtubules appear. This study reveals a multi-step process for cilia cap formation in Tetrahymena.

Keywords:
AFMTetrahymenacap assemblycilia assemblyre-ciliation.

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Area of Science:

  • Cell Biology
  • Biophysics
  • Microscopy

Background:

  • Cilia are vital organelles with motility and sensory roles.
  • Cilia defects cause human ciliopathies.
  • Early cilium assembly, particularly cap formation, is poorly understood.

Purpose of the Study:

  • Investigate the early steps of cilia assembly.
  • Determine the timing and location of cilia cap component assembly.
  • Elucidate the mechanism of cilia cap formation in Tetrahymena.

Main Methods:

  • Atomic Force Microscopy (AFM) in tapping mode.
  • Nanometer-scale resolution with minimal sample manipulation.
  • Observation of Tetrahymena cilia assembly.

Main Results:

  • Cilia assembly requires transient, three-component structures assembled asymmetrically on elongating axonemes.
  • The central pair of microtubules was absent in small, uncapped axonemes.
  • Cilia cap assembly is a multi-step process involving sequential addition of structures.

Conclusions:

  • Propose a model for Tetrahymena cilia cap assembly.
  • Suggest that the cilia cap influences axonemal microtubule polymerization rates.
  • Provide new insights into the fundamental process of cilia biogenesis.