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Nathalie Spassky

Cilia biology and neurogenesis

Multiciliated cells are epithelial cells that line the airways, the oviduct and the brain ventricles. Each of these cells extend multiple (more than 50) long beating cilia producing a constant extracellular flow that clears mucus from the airways, moves ova from the oviducts toward the uterus and propels cerebrospinal fluid (CSF) through the cerebral ventricles. Each beating cilium grows from a modified centriole, also called a basal body. Defects in motile ciliation or cilia number are associated to severe pathologies including neurodevelopmental disorders, irreversible lung failure and sterility by perturbing cilia-based flows.

Nathalie Spassky

Our projects focus on understanding how brain multiciliated cells –called ependymal cells- develop. Their unique location, morphology and function strongly suggest that they actively contribute to the formation and maintenance of neural circuits. We are using a multidisciplinary approach involving mouse molecular genetics, bioinformatic tools, biophysical approaches, ex-vivo culture systems and advanced live cell imaging to investigate (i) how progenitors are specified in ependymal cells; (ii) how cilia are formed and oriented in these cells and iii) how ependymal cells contribute to ventricular morphogenesis and adult neurogenesis.

Alice Meunier

To assess the mechanism of centriole amplification in MCC, we have developed a primary cell culture assay and combined it with single cell high resolutive imaging techniques. We have shown that new centrioles are mainly generated from the single immature daughter centriole and that centriole amplification is driven by a calibrated version of the mitotic oscillator. We now aim to characterize with sub-micrometer precision and in three dimensions the cradle of centriole production, using state of the art imaging techniques. In parallel, a functional approach is run to identify how the mitotic oscillator can be calibrated to control centriole amplification in differentiating MCC without triggering unscheduled and potentially harmfull divisions.

Nathalie Delgehyr

We aim to dissect the cellular mechanisms of the ependymal cell differentiation and maintenance. We notably investigate how the cytoskeleton re-organises during differentiation and what these re-organisations trigger. To keep the complex 3D-environment and architecture of the ependymal cells, we mainly perform our study in-vivo by expressing or depleting key factors directly in the developing brain. We have recently shown that the ciliary beating of ependymal motile cilia is sufficient to induce an actin enrichment around centrioles that in turn protect centrioles against the shear stress generated by the cilia movement. We now characterise earlier cytoskeletal changes and assess their direct contribution in the commitment and organisation of ependymal cells.

Mahuzier A, Shihavuddin A, Fournier C, Lansade P, Faucourt M, Menezes N, Meunier A, Garfa-Traoré M, Carlier MF, Voituriez R, Genovesio A, Spassky N, Delgehyr N. (2018) Ependymal cilia beating induces an actin network to protect centrioles against shear stress. Nat Commun. 9(1):2279.
Pdf available at https://rdcu.be/VCZC

Al Jord A, Shihavuddin A, Servignat d’Aout R, Faucourt M, Genovesio A, Karaiskou A, Sobczak-Thépot J, Spassky N and Meunier A. (2017) Calibrated mitotic oscillator drives motile ciliogenesis. Science 358 (6364):803-806.
Pdf available at http://science.sciencemag.org/cgi/content/full/science.aan8311?ijkey=ZgdxIMo7AEnVM&keytype=ref&siteid=sci

A. Shihavuddin, S. Basu, E. Rexhepaj, F. Delestro, N. Menezes, SM. Sigoillot, E. del Nery, F. Selimi, N. Spassky and A. Genovesio. (2017) Smooth 2D manifold extraction from 3D image stack. Nature Communications. 31(8):15554.

N. Spassky and A. Meunier. (2017) The development and functions of multiciliated epithelia. Nat Rev Mol Cell Biol. 18(7):423-436. Review

P. Foerster, M. Daclin, S. Asm, M. Faucourt, A. Boletta, A. Genovesio and N. Spassky. (2017) mTORC1 signaling and primary cilia are required for brain ventricle morphogenesis. Development. 144(2):201-210.

A. Meunier and N. Spassky. (2016) Centriole continuity: out with the new, in with the old. Curr Opin Cell Biol. 38:60-7. Review

A. Jourdon, A. Gresset, N. Spassky, P. Charnay, P. Topilko, and R. Santos. (2016) Prss56, a novel marker of adult neurogenesis in the mouse brain. Brain Struct Funct. 221(9):4411-4427.

A. Al Jord, N. Spassky, A. Meunier. (2015) Centriole amplification ? #DaughterCentriole. Med/Sci 31(3) :250-3. Review

N. Delgehyr, A. Meunier, M. Faucourt, M. Bosch Grau, L. Strehl, C. Janke and N. Spassky. (2015) Ependymal cell differentiation, from monociliated to multiciliated cells. Methods Cell Biol 127 :19-35.

N. Delgehyr & N. Spassky. Interplay between primary cilia and cell cycle. (2014) Med/Sci 30(11) :976-9. Review

A. Al Jord, A.I. Lemaitre, N. Delgehyr, M. Faucourt, N. Spassky# and A. Meunier#. (2014) Centriole amplification by mother and daughter centrioles differs in multiciliated cells. Nature 516(7529)104-7 (co-corresponding and co-last author).

M.B. Grau, G. Gonzalez Curto, C. Rocha, M.M. Magiera, P. Marques, T. Giordano, N. Spassky# and C. Janke#. (2013) Tubulin glycylases and glutamylases have distinct functions in stabilization and motility of ependymal cilia. J Cell Biol 202(3):441-51. (co-corresponding and co-last author)

A. Meunier, K. Sawamoto, and N. Spassky. (2013) Ependyma, choroid. Chapter 86 : Comprehensive Developmental Neuroscience. D. Rowitch and A. Alvarez-Buylla editors. Elsevier.

N. Spassky. (2013). Motile cilia and brain functions : ependymal motile cilia development, organization, function and their associated pathologies. K. Tucker and T. Caspary editors, Springer.

A. Aguilar, A. Meunier, L. Strehl, J. Martinovic, M. Bonniere, T. Attie-Bitach, F. Encha-Razavi and N. Spassky. (2012). Analysis of human samples reveals impaired Shh-dependent cerebellar development in Joubert syndrome/Meckel syndrome. PNAS 109(42) :16951-6.

B. Guirao, A. Meunier, S. Mortaud, A. Aguilar, J.M. Corsi, L. Strehl, Y. Hirota, A. Desoeuvre, C. Boutin, Y.G. Han, Z. Mirzadeh, H. Cremer, M. Montcouquiol, K. Sawamoto, and N. Spassky. (2010). Coupling between hydrodynamic forces and planar cell polarity orients mammalian motile cilia. Nat Cell Biol, 12(4) :341-50.

Collaborations:

J.P. Baudoin, L. Viou, P. S. Launay, C. Luccardini, S. Espeso, V. Kiyasova, T. Irinopoulou, C. Alvarez, J.P. Rio, T. Boudier, J.P. Lechaire, N. Kessaris, N. Spassky, and C. Métin (2012). Tangentially migrating neurons assemble a primary cilium that promotes their re-orientation to the cortical plate. Neuron, 76(6) :1108-22.

G. Keryer, JR. Pineda, G. Liot, J. Kim, P. Dietrich, C. Benstaali, K. Smith, FP. Cordelières, N. Spassky, RJ. Ferrante, I. Dragatsis, and F. Saudou. (2011). Ciliogenesis is regulated by a huntingtin-HAP1-PCM1 pathway and is altered in Huntington disease. J Clin Invest 121(11) :4372-82.

Y. Hirota, A. Meunier, S. Huang, T. Shimozawa, O. Yamada, Y.S. Kida, M. Inoue, T. Ito, H. Kato, M. Sakaguchi, T. Sunabori, M.A. Nakaya, S. Nonaka, T. Ogura, H. Higuchi, H. Okano, N. Spassky, and K. Sawamoto. (2010). Planar polarity of multiciliated ependymal cells involves the anterior migration of basal bodies regulated by non-muscle myosin II. Development 137(18):3037-46.

A. Molla-Herman, R. Ghossoub, T. Blisnick, A. Meunier, C. Serees, F. Silbermann, C.D. Emmerson, K. Romeo, P. Bourdoncle, A. Schmitt, S. Saunier, N. Spassky, P. Bastin, and A. Benmerah. (2010). The ciliary pocket: an endocytic membrane domain at the base of primary and motile cilia. J Cell Sci, 123 :1785-95.




Primary cell culture of brain multiciliated cells stained for (...)
Primary cell culture of brain multiciliated cells stained for polyglutamylated microtubules in green and tight junctions in red
(p,q) Schematic views of Cen2GFP adult brain ependymal cells from a study (...)
(p,q) Schematic views of Cen2GFP adult brain ependymal cells from a study on the beating direction of their cilia. (r) 3D reconstruction where nuclei (blue), ZO1 cell–cell junctions (red) and centrioles (green) were stained. From Shihavuddin et al., Nat. Com. 2017
Once amplified, procentrioles detach from their centrosome and deuterosome (...)
Once amplified, procentrioles detach from their centrosome and deuterosome platforms along the nuclear membrane.
When Cdk1 activity is disinhibited, post-mitotic multiciliated progenitors (...)
When Cdk1 activity is disinhibited, post-mitotic multiciliated progenitors undergo an abberant mitosis. From Al Jord et al., Science 2017
Double immunostaining of an E14.5 Centrin-2-GFP control cortical (...)
Double immunostaining of an E14.5 Centrin-2-GFP control cortical ventricular surface with Arl13b (cilia, cyan), and phosphorylated S6RP (p-S6RP, magenta) antibodies showing p-S6RP staining at the mother centriole of the centrosome in ciliated cells. From Foerster et al., Development 2017