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Analysis of Neural Crest Cell Fate During Cardiovascular Development Using Cre-Activated lacZ/β-Galactosidase Staining

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Cardiovascular Development

Part of the book series: Methods in Molecular Biology ((MIMB,volume 843))

Abstract

It is important to identify the mechanisms regulating cardiovascular development. However, complex genetic tools are often required, including transgenic animals that express the lacZ transgene encoding the β-galactosidase enzyme under the control of a specific promoter or following recombination with the Cre recombinase. The latter can be useful for identifying specific cell populations of the developing cardiovascular system, including neural crest cells. The tracking of these cells can help clarify their fate in mutant embryos and elucidate the etiology of some congenital cardiovascular birth defects. This chapter highlights the methods used to stain embryonic tissues in whole mount or sections to detect the expression of the lacZ transgene with a focus on tracking cardiac neural crest cells using the Wnt1-Cre and R26R mouse lines. We also provide a protocol using fluorescence-activated cell sorting for collecting neural crest cells for further analysis. These protocols can be used with any embryos expressing Cre and lacZ.

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Acknowledgments

The authors would like to thank Ms. Jeanne Santa Cruz for editing the manuscript and Dr. Chantale Lacelle for helping with the FACS. This work was supported by the NIH/ NIDCR U24 DE16472 (LBR) and T32 DE018380 (YZ) and a Research Development Grant (LBR) from the Office of the Vice President for Research & Graduate Studies/Texas A&M Health Science Center.

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Authors and Affiliations

  1. Department of Biomedical Sciences, Texas A&M Healthy Science Center-Baylor College of Dentistry, Dallas, TX, USA

    Yanping Zhang & L. Bruno Ruest

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  1. Yanping Zhang
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  2. L. Bruno Ruest
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Correspondence to L. Bruno Ruest .

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Editors and Affiliations

  1. College of Medicine, Dept. Systems Biology &, Texas A&M Health Science Center, SW. HK Dodgen Loop 702, Temple, 76504-7105, Texas, USA

    Xu Peng

  2. School of Veterinary Medicine, Department of Molecular Medicine, Cornell University, Ithaca, 14853, New York, USA

    Marc Antonyak

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Zhang, Y., Ruest, L.B. (2012). Analysis of Neural Crest Cell Fate During Cardiovascular Development Using Cre-Activated lacZ/β-Galactosidase Staining. In: Peng, X., Antonyak, M. (eds) Cardiovascular Development. Methods in Molecular Biology, vol 843. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-523-7_12

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  • DOI: https://doi.org/10.1007/978-1-61779-523-7_12

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-522-0

  • Online ISBN: 978-1-61779-523-7

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