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Intron-Specific Neuropeptide Probes

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Neuropeptides

Part of the book series: Methods in Molecular Biology ((MIMB,volume 789))

Abstract

Measurements of changes in pre-mRNA levels by intron-specific probes are generally accepted as more closely reflecting changes in gene transcription rates than are measurements of mRNA levels by exonic probes. This is, in part, because the pre-mRNAs, which include the primary transcript and various splicing intermediates located in the nucleus (also referred to as heteronuclear RNAs, or hnRNAs), are processed rapidly (with half-lives <60 min) as compared to neuropeptide mRNAs, which are then transferred to the cytoplasm and which have much longer half-lives (often over days). In this chapter, we describe the use of exon-and intron-specific probes to evaluate oxytocin (OT) and vasopressin (VP) neuropeptide gene expression by analyses of their mRNAs and hnRNAs by quantitative in situ hybridization (qISH) and also by using specific PCR primers in quantitative, real-time PCR (qPCR) procedures.

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Acknowledgments

We wish to thank various past and present members in our LNC, NINDS, NIH laboratory, Noriko Mutsuga, Ray Fields, Shirley House, Daniel Lubelski, and Madison Stevens, and colleagues in other NIH institutes, Eva Mezey, Sharon Key and Zuszanna Toth in the NICDR, and W. Scott Young in NIMH for their help in establishing the qISH and qPCR methods in our laboratory. This work was supported by the intramural program of the NIH, NINDS.

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Authors and Affiliations

  1. Laboratory of Neurochemistry, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD, USA

    Harold Gainer & Todd A. Ponzio

  2. Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China

    Chunmei Yue

  3. Department of Orthopaedics, School of Medicine, University of Occupational and Environmental Health, Kitakyushu-City, Fukuoka, Japan

    Makoto Kawasaki

Authors
  1. Harold Gainer
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  2. Todd A. Ponzio
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  3. Chunmei Yue
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  4. Makoto Kawasaki
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Corresponding author

Correspondence to Harold Gainer .

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Editors and Affiliations

  1. , Dipt. di Morfofisiologia Veterinaria, Università degli Studi di Torino, Via Leonardo da Vinci 44, Grugliasco, 10095, Torino, Italy

    Adalberto Merighi

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Gainer, H., Ponzio, T.A., Yue, C., Kawasaki, M. (2011). Intron-Specific Neuropeptide Probes. In: Merighi, A. (eds) Neuropeptides. Methods in Molecular Biology, vol 789. Humana Press. https://doi.org/10.1007/978-1-61779-310-3_5

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  • DOI: https://doi.org/10.1007/978-1-61779-310-3_5

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-309-7

  • Online ISBN: 978-1-61779-310-3

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