| Immunohistochemistry of Kidney a-SMA, Collagen 1, and Collagen 3, in A Novel Mouse Model of Reno-cardiac Syndrome
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Author:
Date:
2020-09-20
[Abstract] Cardiorenal syndrome defines a synergistic pathology of the heart and kidneys where failure of one organ causes failure in the other. The incidence of cardiovascular mortality caused by this syndrome, is 20 fold higher in the end stage renal disease (ESRD) population compared to the population as a whole thus necessitating the need for improved therapeutic strategies to combat reno-cardiac pathologies.
Murine in vivo models play a major role in such research permitting precise genetic modification thus reducing miscellany, however presently there is no steadfast model of reno-cardiac syndrome in the most common genetically modified mouse strain, the C57BL/6 mouse. In this study we have modified an established model of chronic renal disease using adenine diet and ...
[摘要] [摘要 ] 心肾综合征定义了心脏和肾脏的协同病理，其中一个器官的衰竭导致另一个器官的衰竭。与整个人群相比，该综合征导致的心血管疾病死亡率在终末期肾脏病（ESRD）人群中要高出20倍，因此有必要改善治疗策略以应对肾病。
小鼠体内模型在允许精确基因修饰从而减少杂项的研究中起主要作用，但是目前在最常见的基因修饰小鼠品系C57BL / 6小鼠中还没有稳定的雷诺-心脏综合征模型。在这项研究中，我们使用腺嘌呤饮食修改了已建立的慢性肾脏疾病模型，并扩展了在C57BL / 6小鼠中实现慢性肾功能衰竭和随之而来的肾心脏综合征的相关病理。
使八周大的雄性C57BL / 6小鼠适应7天，然后给予0.15％腺嘌呤饮食或对照饮食20周，此后终止实验，收集血液，尿液和器官并进行生化和免疫组织化学分析。
施用0.15％的腺嘌呤饮食会导致进行性肾功能衰竭，从而导致肾性心脏病综合征，这可通过心体重比显着增加来证实（P <0.0001）。血液生化表明，用腺嘌呤喂养的小鼠血清肌酐，尿素含量显着增加（P <0.0001），肾小球滤过率显着降低（P <0.05），而肾脏的α-SMA，胶原蛋白1和胶原蛋白3免疫组化显示严重的纤维化。
我们提出了一种新型的腺嘌呤饮食方案，该方案在C57BL / ...
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| Preparation of Sequencing RNA Libraries through Chemical Cross-linking Coupled to Affinity Purification (cCLAP) in Saccharomyces cerevisiae
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Author:
Date:
2018-10-05
[Abstract] Ribonucleoprotein particles (mRNPs) are complexes consisting of mRNAs and RNA-binding proteins (RBPs) which control mRNA transcription localization, turnover, and translation. Some mRNAs within the mRNPs have been shown to undergo degradation or storage. Those transcripts can lack general mRNA elements, like the poly(A) tail or 5’ cap structure, which prevent their identification through the application of widely-used approaches like oligo(dT) purification. Here, we describe a modified cross-linking affinity purification protocol (cCLAP) based on existing cross-linking and immunoprecipitation (CLIP) methods to isolate mRNAs which could be deadenylated, decapped and/or partially degraded in mRNPs, opening the possibility to detect different types of non-coding RNAs (ncRNAs). Once isolated, ...
[摘要] 核糖核蛋白颗粒（mRNP）是由mRNA和RNA结合蛋白（RBP）组成的复合物，其控制mRNA转录定位，转换和翻译。已显示mRNP内的一些mRNA经历降解或储存。那些转录物可能缺乏一般的mRNA元件，如poly（A）尾或5'帽结构，这通过应用广泛使用的方法如oligo（dT）纯化来阻止它们的鉴定。在这里，我们描述了基于现有的交联和免疫沉淀（CLIP）方法的修饰的交联亲和纯化方案（cCLAP），以分离mRNP中可被去腺苷酸化，去除和/或部分降解的mRNA，从而开启了检测不同的可能性。非编码RNA（ncRNA）的类型。分离后，将RNA进行衔接子连接，然后进行下一代测序（NGS）。由于快速有效的交联和淬灭步骤，该方案也适用于瞬时诱导的mRNP颗粒。实例包括由外在应激物触发的处理体（PB）或应力颗粒（SG）。其重现性和广泛应用使该方案成为研究特定RNP的RNA组成的有用且有力的工具。
【背景】mRNP内转录物的表征对于理解细胞转录和转录后过程至关重要。通过交联和免疫沉淀，然后通过RNA-Seq从mRNP颗粒中分离RNA已经成为鉴定mRNA靶标的常用方法（Tagwerker et al。，2006; Hafner et al。，2010; Kishore et al。，2011）。 ...
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