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Cell type-specific expression of endogenous cardiac Troponin I antisense RNA in the neonatal rat heart

机译:新生大鼠心脏内源性心肌肌钙蛋白I反义RNA的细胞类型特异性表达

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Since the number of detected natural antisense RNA is growing, investigations upon the expression pattern of the antisense RNA become more important. As we focused our work on natural occurring antisense transcripts in human and rat heart tissues, we were interested in the question, whether the expression pattern of antisense and sense RNA can vary in different cell types of the same tissue. In our previous analysis of total neonatal rat heart tissue, we demonstrated the co-expression of both cTnI RNA species in this tissue. Now we investigated the expression of antisense and sense RNA quantitatively in neonatal cardiomyocytes (NCMs) and neonatal cardiac fibroblasts (NCFs). Performing northern blot as well as RT-PCR, we could detect natural antisense and sense RNA transcripts of cTnI in NCM and NCF implying that these transcripts are co-expressed in both cell types. The absolute amounts of the RNA transcripts were higher in NCM. Both RNA species showed identical sizes in the northern blot. Quantification by real-time PCR revealed a higher relative level of natural antisense RNA in NCF compared to NCM which points out to a cell type-specific expression of sense and antisense RNA. Our observations suggest that antisense RNA transcription may contribute to a cell type-specific regulation of the cTnI gene.
机译:由于检测到的天然反义RNA的数目正在增加,因此对反义RNA的表达模式的研究变得更加重要。当我们将工作重点放在人和大鼠心脏组织中的天然反义转录本上时,我们对这个问题感兴趣,即反义和有义RNA的表达模式是否可以在同一组织的不同细胞类型中变化。在我们之前对新生大鼠心脏组织的总分析中,我们证明了两种cTnI RNA物种在该组织中的共表达。现在我们定量研究了新生儿心肌细胞(NCM)和新生儿心脏成纤维细胞(NCF)中反义和有义RNA的表达。进行northern blot和RT-PCR,我们可以在NCM和NCF中检测cTnI的天然反义和有义RNA转录本,这意味着这些转录本在两种细胞类型中都共同表达。在NCM中,RNA转录本的绝对量更高。两种RNA物种在northern印迹中均显示相同的大小。实时PCR定量显示,与NCM相比,NCF中天然反义RNA的相对水平更高,这表明有义和反义RNA的细胞类型特异性表达。我们的观察结果表明,反义RNA转录可能有助于cTnI基因的细胞类型特异性调控。

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