Cilia visualization in Xenopus cells

Multi-ciliated cells (MCCs) are terminally differentiated epithelia that are present in all metazoans and many unicellular eukaryotes1,2. In marine organisms they play a key role in locomotion and feeding3, while in mammals they clear mucus from the lungs, circulate cerebrospinal fluid in the nervous system and transport eggs and sperm in the reproductive tracts2.

Individual MCCs contain hundreds of motile cilia that rest on modified centrioles called basal bodies4. MCCs are polarized relative to each in a tissue, and individual cilia are polarized relative to each other within the MCC5. Individual cilia beat, which generates a unidirectional fluid flow along the planar axis of the tissue6. Abnormalities in cilia abundance, orientation and/or beating have severe implications for human health, causing chronic respiratory infections, hydrocephalus and male infertility7.

Recent advances in ‘omic’ methodologies and in live cell imaging techniques has renewed interest in understanding MCCs2. Most studies use the African clawed frog (Xenopus laevis) as a model system in cilia research4,5. Here, MCCs were obtained by in vitro differentiation of Xenopus-derived cell culture and were imaged using Nanolive’s 3D Cell Explorer. Images were acquired for 4 mins at an acquisition frequency of 1 image every 2 secs.

Information on Z axis (depth) was processed so that a color scale (a gradient of color ranging from blue to pink) was applied to it, providing a sense of spacial organization in that axis. Full cells or cell components closer to the dish surface were colored blue, while pink accounted for cells or cellular content positioned further from the dish surface.

Sample courtesy of Camille Boutin, IBDM Marseille.

References

  1. Brooks, E. R. & Wallingford, J. B. Multiciliated Cells. Curr. Biol. 24, R973–R982 (2014).
  2. Spassky, N. & Meunier, A. The development and functions of multiciliated epithelia. Nat. Rev. Mol. Cell Biol. 18, 423–436 (2017).
  3. Marinković, M., Berger, J. & Jékely, G. Neuronal coordination of motile cilia in locomotion and feeding. Philos. Trans. R. Soc. Lond. B. Biol. Sci. 375, 20190165 (2020).
  4. Boutin, C. & Kodjabachian, L. Biology of multiciliated cells. Curr. Opin. Genet. Dev. 56, 1–7 (2019).
  5. Werner, M. E. & Mitchell, B. J. Understanding ciliated epithelia: The power of Xenopus. Genesis 50, 176–185 (2012).
  6. Satir, P., Heuser, T. & Sale, W. S. A structural basis for how motile cilia beat. Bioscience 64, 1073–1083 (2014).
  7. Bisgrove, B. W. & Yost, H. J. The roles of cilia in developmental disorders and disease. Development 133, 4131–4143 (2006).

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