Abstract
Whole mount in situ hybridization is a sensitive method used to characterize the spatial and temporal expression of RNA transcripts throughout an entire tissue. This method is an excellent tool for studying gene expression during embryonic development. Here, we describe a procedure for digoxigenin labeled in situ hybridization on whole embryos.
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References
Gall JG, Pardue ML (1969) Formation and detection of RNA-DNA hybrid molecules in cytological preparations. Proc Natl Acad Sci U S A 63(2):378–383
Riddle RD, Johnson RL, Laufer E, Tabin C (1993) Sonic hedgehog mediates the polarizing activity of the ZPA. Cell 75(7):1401–1416
Zakany J, Duboule D (1993) Correlation of expression of Wnt-1 in developing limbs with abnormalities in growth and skeletal patterning. Nature 362(6420):546–549
Chen H, Johnson RL (1999) Dorsoventral patterning of the vertebrate limb: a process governed by multiple events. Cell Tissue Res 296(1):67–73
Cohn MJ, Izpisua-Belmonte JC, Abud H, Heath JK, Tickle C (1995) Fibroblast growth factors induce additional limb development from the flank of chick embryos. Cell 80(5):739–746
Lettice LA, Hill RE (2005) Preaxial polydactyly: a model for defective long-range regulation in congenital abnormalities. Curr Opin Genet Dev 15(3):294–300
McGlinn E, van Bueren KL, Fiorenza S, Mo R, Poh AM, Forrest A et al (2005) Pax9 and Jagged1 act downstream of Gli3 in vertebrate limb development. Mech Dev 122(11):1218–1233
Sheth R, Gregoire D, Dumouchel A, Scotti M, Pham JM, Nemec S et al (2013) Decoupling the function of Hox and Shh in developing limb reveals multiple inputs of Hox genes on limb growth. Development 140(10):2130–2138
Tabin C, Wolpert L (2007) Rethinking the proximodistal axis of the vertebrate limb in the molecular era. Genes Dev 21(12):1433–1442
Zuniga A, Haramis AP, McMahon AP, Zeller R (1999) Signal relay by BMP antagonism controls the SHH/FGF4 feedback loop in vertebrate limb buds. Nature 401(6753):598–602
Acloque H, Wilkinson DG, Nieto MA (2008) In situ hybridization analysis of chick embryos in whole-mount and tissue sections. Methods Cell Biol 87:169–185
Wilkinson DG, Nieto MA (1993) Detection of messenger RNA by in situ hybridization to tissue sections and whole mounts. Methods Enzymol 225:361–373
Neufeld SJ, Zhou X, Vize PD, Cobb J (2013) mRNA fluorescence in situ hybridization to determine overlapping gene expression in whole-mount mouse embryos. Dev Dyn 242(9):1094–1100
Wilkinson DG (1995) RNA detection using non-radioactive in situ hybridization. Curr Opin Biotechnol 6(1):20–23
Komminoth P (1992) Digoxigenin as an alternative probe labeling for in situ hybridization. Diagn Mol Pathol 1(2):142–150
Acknowledgments
This work was supported by NIH/NIAMS grants (A032092 and A032121) to M.J.H and (AR071967 and AR076325) to C.M.K.
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Sharma, D., Hilton, M.J., Karner, C.M. (2021). Whole Mount In Situ Hybridization in Murine Tissues. In: Hilton, M.J. (eds) Skeletal Development and Repair. Methods in Molecular Biology, vol 2230. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1028-2_22
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DOI: https://doi.org/10.1007/978-1-0716-1028-2_22
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