Abstract
In vivo lineage tracing is a valuable technique to study cellular behavior. Our lab developed a lineage tracing method, based on the Cre/lox system, to genetically induce clonal labelling of cells and follow their progeny. Here we describe a protocol for temporally controlled clonal labelling and for microdissection of individual mouse hair follicles. We further present staining and visualization techniques used in our lab to analyze clones issued from genetically induced labelling.
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References
Fuchs E (2009) The tortoise and the hair: slow-cycling cells in the stem cell race. Cell 137:811–819
Legué E, Sequeira I, Nicolas J-F (2012) Hair follicle stem cells. In: Hayat MA (ed) Stem cells and cancer stem cells, vol 3. Springer, New York, pp 35–47
Fuchs E, Horsley V (2008) More than one way to skin. Genes Dev 22:976–985
Legué E, Sequeira I, Nicolas J-F (2010) Hair follicle renewal: authentic morphogenesis that depends on a complex progression of stem cell lineages. Development 137:569–577
Millar SE (2002) Molecular mechanisms regulating hair follicle development. J Investig Dermatol 118:216–225
Müller-Röver S, Handjiski B, van der Veen C et al (2001) A comprehensive guide for the accurate classification of murine hair follicles in distinct hair cycle stages. J Investig Dermatol 117:3–15
Stenn KS, Paus R (2001) Controls of hair follicle cycling. Physiol Rev 81:449–494
Legué E, Nicolas J-F (2005) Hair follicle renewal: organization of stem cells in the matrix and the role of stereotyped lineages and behaviors. Development 132:4143–4154
Feil R, Brocard J, Mascrez B et al (1996) Ligand-activated site-specific recombination in mice. Proc Natl Acad Sci U S A 93:10887–10890
Soriano P (1999) Generalized lacZ expression with the ROSA26 Cre reporter strain. Nat Genet 21:70–71
Vooijs M, Jonkers J, Berns A (2001) A highly efficient ligand-regulated Cre recombinase mouse line shows that LoxP recombination is position dependent. EMBO Reports 2:292–297
Metzger D, Chambon P (2001) Site- and time-specific gene targeting in the mouse. Methods 24:71–80, San Diego, Calif
Indra AK, Warot X, Brocard J et al (1999) Temporally-controlled site-specific mutagenesis in the basal layer of the epidermis: comparison of the recombinase activity of the tamoxifen-inducible Cre-ER(T) and Cre-ER(T2) recombinases. Nucleic Acids Res 27:4324–4327
Jaks V, Barker N, Kasper M et al (2008) Lgr5 marks cycling, yet long-lived, hair follicle stem cells. Nat Genet 40:1291–1299
Snippert HJ, Haegebarth A, Kasper M et al (2010) Lgr6 marks stem cells in the hair follicle that generate all cell lineages of the skin. Science 327:1385–1389
Li M, Indra AK, Warot X et al (2000) Skin abnormalities generated by temporally controlled RXRalpha mutations in mouse epidermis. Nature 407:633–636
Kawamoto S, Niwa H, Tashiro F et al (2000) A novel reporter mouse strain that expresses enhanced green fluorescent protein upon Cre-mediated recombination. FEBS Lett 470:263–268
Srinivas S, Watanabe T, Lin CS et al (2001) Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus. BMC Dev Biol 1:4
Muzumdar MD, Tasic B, Miyamichi K et al (2007) A global double-fluorescent Cre reporter mouse, genesis 45:593–605
Livet J, Weissman TA, Kang H et al (2007) Transgenic strategies for combinatorial expression of fluorescent proteins in the nervous system. Nature 450:56–62
Snippert HJ, van der Flier LG, Sato T et al (2010) Intestinal crypt homeostasis results from neutral competition between symmetrically dividing Lgr5 stem cells. Cell 143:134–144
Sequeira I, Nicolas J-F (2012) Redefining the structure of the hair follicle by 3D clonal analysis. Development 139:3741–3751
Bonnerot C, Nicolas JF (1993) Clonal analysis in the intact mouse embryo by intragenic homologous recombination, Comptes rendus de l'Académie des sciences, Série III. Sciences de la vie 316:1207–1217
Acknowledgments
This work was supported by the Institut Pasteur and by the French Agence National de la Recherche n°10-01 (DEV-Process to I.S. and J.F.N). All experiments were carried out in accordance with the national guidelines for care and use of laboratory animals.
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Sequeira, I., Legué, E., Capgras, S., Nicolas, JF. (2013). Microdissection and Visualization of Individual Hair Follicles for Lineage Tracing Studies. In: Turksen, K. (eds) Epidermal Cells. Methods in Molecular Biology, vol 1195. Springer, New York, NY. https://doi.org/10.1007/7651_2013_48
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DOI: https://doi.org/10.1007/7651_2013_48
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Print ISBN: 978-1-4939-1223-0
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