Intron-specific neuropeptide probes.
Intron-specific neuropeptide probes.
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内含子特异性神经肽探针。
DOI：
10.1007/978-1-61779-310-3_5
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
Kawasaki, Makoto
中科院分区：
文献类型：
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作者：
Gainer, Harold;Ponzio, Todd A;Yue, Chunmei;Kawasaki, Makoto
通过内含子特异性探针测量前mRNA水平的变化通常被认为比通过外显子探针测量mRNA水平更能反映基因转录速率的变化。这部分是因为前mRNA，包括初级转录物和位于细胞核中的各种剪接中间体（也称为异源RNA或hnRNA），与神经肽mRNA相比被快速加工（半衰期<60分钟），神经肽mRNA然后转移到细胞质并且具有长得多的半衰期（通常超过数天）。在本章中，我们描述了使用外显子和内含子特异性探针来评估催产素（OT）和加压素（VP）神经肽基因的表达，通过定量原位杂交（qISH）分析它们的mRNA和hnRNA，并通过使用特异性PCR引物进行定量，实时PCR（qPCR）程序。
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.