Detection of apoptotic cells based on in situ hybridization chain reaction using specific hairpins
Detection of apoptotic cells based on in situ hybridization chain reaction using specific hairpins
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使用特定发夹的原位杂交链式反应检测凋亡细胞
DOI：
10.1007/s10495-022-01782-5
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发表时间：
2022-11
期刊：
影响因子：
7.2
通讯作者：
Hongbin Yuan
中科院分区：
文献类型：
--
作者：
Mei Yang;Ruihua Ji;Zhengqing Zhao;Wenwen Wang;Ye Lu;Zhenghua Xiang;Hongbin Yuan
越来越多的实验研究细胞程序性死亡/凋亡，其特征之一是DNA片段化。目前唯一的原位检测DNA片段化的方法是末端脱氧核苷酸转移酶介导的dUTP缺口末端标记，TUNEL。本研究建立了一种新的原位检测凋亡DNA片段的方法，即原位杂交链反应（isHCR）。该测定的原理是凋亡细胞DNA片段的粘性末端序列非特异性地启动特异性检测凋亡细胞的杂交链反应。TUNEL法和isHCR法联合检测的结果表明，绝大多数isHCR阳性细胞也被TUNEL标记。原位HCR常能检测到经典TUNEL法所不能检测到的胞浆中的DNA片段，这些细胞可能处于凋亡的早期阶段。它还表明，DNA片段转移到细胞质中的细胞凋亡过程中。因为染色过程不需要末端脱氧核苷酸转移酶作为TUNEL染色，isHCR染色成本低，可以在大量的组织标本上进行。据信isHCR具有原位检测凋亡细胞的DNA片段化的潜力。
There are an increasing number of experiments to study programmed cell death/apoptosis, one of the characteristics of which is DNA fragmentation. The only current method for in situ detection of DNA fragmentation is Terminal deoxynucleotidyl transferase mediated-dUTP Nick End Labeling, TUNEL. In this study, a new method for in situ detection of apoptotic DNA fragments, namely In Situ Hybridization Chain Reaction, isHCR, was established. The principle of the assay is that the sticky end sequence of the apoptotic cell DNA fragment non-specifically initiates a hybridization chain reaction that specifically detects the apoptotic cell. The results of the combined TUNEL and isHCR method demonstrated that the majority of isHCR-positive cells were also labeled by TUNEL. In situ HCR often detect DNA fragments in the cytoplasm that the classical TUNEL method couldnot, and these cells may be in the early stages of apoptosis. It also indicates that DNA fragments are transferred to the cytoplasm during apoptosis. Because the staining process does not require terminal deoxynucleotidyl transferase as TUNEL staining does, isHCR staining cost low and can be performed on a large number of tissue specimens. It is believed that isHCR has the potential to detect DNA fragmentation of apoptotic cells in situ.
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影响因子：
2.2
作者：
LOCKSHIN, RICHARD A.;WILLIAMS, CARROLL M.
通讯作者：
WILLIAMS, CARROLL M.
影响因子：
--
作者：
F. Li;Y. Zhang;S. L. Ma
通讯作者：
F. Li;Y. Zhang;S. L. Ma
影响因子：
64.8
作者：
Lammert CR;Frost EL;Bellinger CE;Bolte AC;McKee CA;Hurt ME;Paysour MJ;Ennerfelt HE;Lukens JR
通讯作者：
Lukens JR
影响因子：
--
作者：
Nader Maghsoudi;Zahra Zakeri;R. Lockshin
通讯作者：
Nader Maghsoudi;Zahra Zakeri;R. Lockshin
DOI：
10.1083/jcb.119.3.493
发表时间：
1992-11
期刊：
The Journal of cell biology
影响因子：
--
作者：
Gavrieli Y;Sherman Y;Ben-Sasson SA
通讯作者：
Ben-Sasson SA