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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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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