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首页> 外文期刊>Molecular urology >Loss of mitochondrial DNA in rabbit bladder smooth muscle following partial outlet obstruction results from lack of organellar DNA replication
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Loss of mitochondrial DNA in rabbit bladder smooth muscle following partial outlet obstruction results from lack of organellar DNA replication

机译：缺乏出口的细胞器DNA复制导致部分出口阻塞后兔膀胱平滑肌线粒体DNA的丢失

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摘要

When the rabbit bladder outlet is partially obstructed, the relative amount of mitochondrial (mt) DNA per cell in bladder smooth muscle falls rapidly. In order to assess whether this loss of organellar genome results from attenuation of mt DNA replication, we cloned portions of rabbit genes specifying the single-strand binding (SSB) protein required for initiation of mt DNA replication, and the catalytic subunit of DNA polymerase < gamma > (pol < gamma >), the replication enzyme itself. We then designed primer-probe systems for real-time RT-PCR (TaqMan< trademark >) analyses for each gene. These were used to assess mRNA in preparations from bladder smooth muscle and mucosa from rabbits subjected to surgical obstruction of the bladder outlet for up to 14 days. mRNA from the pol < gamma > gene remained essentially at control level in smooth muscle and mucosa in all samples. In mucosa, mRNA from the SSB protein gene remained virtually at control levels in all samples, as did mt genome copy number. In smooth muscle, however, levels of this mRNA declined by >95% within 3 days of obstruction and remained at that level through 14 days; this attenuation of SSB protein mRNA paralleled the loss of mt DNA in the same smooth muscle samples. Thus, lack of mt SSB protein, and consequently attenuated mt DNA replication, is a primary factor in the loss of mt genome copies in bladder smooth muscle after outlet obstruction in the rabbit model of benign bladder dysfunction.
机译：当兔子膀胱出口部分受阻时，膀胱平滑肌中每个细胞的线粒体（mt）DNA的相对数量迅速下降。为了评估这种细胞器基因组损失是否是由于mt DNA复制的减弱所致，我们克隆了兔基因的某些部分，这些序列指定了启动mt DNA复制所需的单链结合（SSB）蛋白，以及DNA聚合酶的催化亚基< gamma>（pol ），即复制酶本身。然后，我们设计了用于每个基因的实时RT-PCR（TaqMan <商标>）分析的引物探针系统。这些被用来评估来自膀胱平滑肌和粘膜的制剂中的mRNA的表达，这些兔子是经过长达14天的膀胱出口手术梗阻的。在所有样品中，pol 基因的mRNA基本上保持在平滑肌和粘膜的对照水平。在粘膜中，所有样品中来自SSB蛋白基因的mRNA几乎保持在对照水平，mt基因组拷贝数也是如此。然而，在平滑肌中，该mRNA的水平在阻塞后3天内下降了> 95％，并在14天内一直保持该水平。 SSB蛋白mRNA的这种减弱与同一平滑肌样品中mt DNA的丢失平行。因此，在兔良性膀胱功能障碍模型中，缺乏mt SSB蛋白，从而导致mt DNA复制减弱，是导致出口阻塞后膀胱平滑肌mt基因组拷贝丢失的主要因素。

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