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Identification and Visualization of Protein Expression in Whole Mouse Embryos by Immunofluorescence

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Epiblast Stem Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2490))

Abstract

Mouse embryo studies are pivotal for the understanding of early development. Analysis of the spatial and temporal changes of protein expression during development of a mouse embryo allows us to identify the genetic basis of errors of development in animal disease models. Immunofluorescence is a powerful technique to study the localization and variation in expression pattern of specific proteins in cells, tissues, and organs. Detecting the antigens with their specific antibodies labeled with fluorescent probes allows visualization of proteins at the cellular level. Here, we provide the optimized protocol of immunostaining whole mouse embryos at embryonic stages E7.5 to E11.5.

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References

  1. Yu SJAD (2017) Basic science methods for clinical researchers. https://doi.org/10.1016/B978-0-12-803077-6.00008-4

    Book  Google Scholar 

  2. Fan X, Masamsetti VP, Sun JQ, Engholm-Keller K, Osteil P, Studdert J, Graham ME, Fossat N, Tam PP (2021) TWIST1 and chromatin regulatory proteins interact to guide neural crest cell differentiation. elife 10:e62873. https://doi.org/10.7554/eLife.62873

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Niu X, Liu L, Wang T, Chuan X, Yu Q, Du M, Gu Y, Wang L (2020) Mapping of extrinsic innervation of the gastrointestinal tract in the mouse embryo. J Neurosci 40(35):6691–6708. https://doi.org/10.1523/JNEUROSCI.0309-20.2020

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Koren E, Yosefzon Y, Ankawa R, Soteriou D, Jacob A, Nevelsky A, Ben-Yosef R, Bar-Sela G, Fuchs Y (2018) ARTS mediates apoptosis and regeneration of the intestinal stem cell niche. Nat Commun 9(1):4582. https://doi.org/10.1038/s41467-018-06941-4

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Adameyko I, Lallemend F, Furlan A, Zinin N, Aranda S, Kitambi SS, Blanchart A, Favaro R, Nicolis S, Lubke M, Muller T, Birchmeier C, Suter U, Zaitoun I, Takahashi Y, Ernfors P (2012) Sox2 and Mitf cross-regulatory interactions consolidate progenitor and melanocyte lineages in the cranial neural crest. Development 139(2):397–410. https://doi.org/10.1242/dev.065581

    Article  PubMed  PubMed Central  Google Scholar 

  6. Rios AC, Capaldo BD, Vaillant F, Pal B, van Ineveld R, Dawson CA, Chen Y, Nolan E, Fu NY, 3DTCLSM Group, Jackling FC, Devi S, Clouston D, Whitehead L, Smyth GK, Mueller SN, Lindeman GJ, Visvader JE (2019) Intraclonal plasticity in mammary tumors revealed through large-scale single-cell resolution 3D imaging. Cancer Cell 35(4):618–632.e6. https://doi.org/10.1016/j.ccell.2019.02.010

    Article  CAS  PubMed  Google Scholar 

  7. Dodt HU, Leischner U, Schierloh A, Jahrling N, Mauch CP, Deininger K, Deussing JM, Eder M, Zieglgansberger W, Becker K (2007) Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain. Nat Methods 4(4):331–336. https://doi.org/10.1038/nmeth1036

    Article  CAS  PubMed  Google Scholar 

  8. Genina EA, Bashkatov AN, Tuchin VV (2010) Tissue optical immersion clearing. Expert Rev Med Devices 7(6):825–842. https://doi.org/10.1586/erd.10.50

    Article  PubMed  Google Scholar 

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Correspondence to V. Pragathi Masamsetti .

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© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Masamsetti, V.P., Tam, P.P.L. (2022). Identification and Visualization of Protein Expression in Whole Mouse Embryos by Immunofluorescence. In: Osteil, P. (eds) Epiblast Stem Cells. Methods in Molecular Biology, vol 2490. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2281-0_4

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  • DOI: https://doi.org/10.1007/978-1-0716-2281-0_4

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2280-3

  • Online ISBN: 978-1-0716-2281-0

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