Radioactive in situ hybridization for detecting diverse gene expression patterns in tissue.

Journal of Visualized Experiments : JoVE
Chun-Chun ChenErich D Jarvis

Abstract

Knowing the timing, level, cellular localization, and cell type that a gene is expressed in contributes to our understanding of the function of the gene. Each of these features can be accomplished with in situ hybridization to mRNAs within cells. Here we present a radioactive in situ hybridization method modified from Clayton et al. (1988)(1) that has been working successfully in our lab for many years, especially for adult vertebrate brains(2-5). The long complementary RNA (cRNA) probes to the target sequence allows for detection of low abundance transcripts(6,7). Incorporation of radioactive nucleotides into the cRNA probes allows for further detection sensitivity of low abundance transcripts and quantitative analyses, either by light sensitive x-ray film or emulsion coated over the tissue. These detection methods provide a long-term record of target gene expression. Compared with non-radioactive probe methods, such as DIG-labeling, the radioactive probe hybridization method does not require multiple amplification steps using HRP-antibodies and/or TSA kit to detect low abundance transcripts. Therefore, this method provides a linear relation between signal intensity and targeted mRNA amounts for quantitative analysis. It allow...Continue Reading

References

Apr 15, 1997·Proceedings of the National Academy of Sciences of the United States of America·E D Jarvis, F Nottebohm
Nov 10, 1998·Neuron·E D JarvisF Nottebohm
Dec 19, 2000·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·A F MoormanV M Christoffels
Apr 2, 2004·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Sebastian HaeslerConstance Scharff
Jul 6, 2004·The Journal of Comparative Neurology·Kazuhiro WadaMasatoshi Hagiwara
Feb 3, 2005·Nature Reviews. Neuroscience·Erich D JarvisUNKNOWN Avian Brain Nomenclature Consortium
Jul 16, 2005·Proceedings of the National Academy of Sciences of the United States of America·Kim L HokeWalter Wilczynski
Jan 28, 2006·The Journal of Comparative Neurology·Chun-Chun Chen, Russell D Fernald
Oct 5, 2006·Proceedings of the National Academy of Sciences of the United States of America·Kazuhiro WadaErich D Jarvis
May 20, 2008·Methods in Cell Biology·Hervé AcloqueM Angela Nieto
Nov 26, 2008·Methods in Molecular Biology·Radma Mahmood, Ivor Mason
Jan 9, 2010·The Journal of Comparative Neurology·Lubica KubikovaErich D Jarvis
May 28, 2010·The Journal of Comparative Neurology·Haruhito HoritaErich D Jarvis

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Citations

Jul 3, 2013·The Journal of Comparative Neurology·Erich D JarvisKazuhiro Wada
Jun 25, 2015·PloS One·Mukta ChakrabortyErich D Jarvis
Oct 1, 2020·Der Pathologe·Christina Schipper, Dirk Zielinski
Jun 17, 2018·International Journal of Obesity : Journal of the International Association for the Study of Obesity·Máté DurstZsuzsanna E Tóth
Mar 23, 2020·Brain Structure & Function·Rita MatuskaZsuzsanna E Tóth
Jul 3, 2013·The Journal of Comparative Neurology·Chun-Chun ChenErich D Jarvis

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