Semi-quantitative immunohistochemical detection of 5-hydroxymethyl-cytosine reveals conservation of its tissue distribution between amphibians and mammals
Semi-quantitative immunohistochemical detection of 5-hydroxymethyl-cytosine reveals conservation of its tissue distribution between amphibians and mammals
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DOI：
10.4161/epi.7.2.18949
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发表时间：
2012-02-01
期刊：
影响因子：
3.7
通讯作者：
Ruzov, Alexey
中科院分区：
文献类型：
--
作者：
Almeida, Rimple D.;Sottile, Virginie;Ruzov, Alexey
5-羟甲基胞嘧啶（5-hmC）是胞嘧啶的一种修饰形式，由于其在转录调控中的潜在作用，近年来引起了人们的广泛关注。根据一些报告，5-羟甲基胞嘧啶分布在哺乳动物中具有组织特异性。因此，5-hmC在胚胎细胞群和成人神经元组织中富集。在这里，我们描述了一种新的半定量免疫组化检测5-hmC的方法，并利用它来评估两栖动物组织中这种修饰的水平。我们发现，类似于哺乳动物胚胎，5-hmC是丰富的蝾螈蝌蚪相比，成人组织。我们的数据表明，5-hmC分布是组织特异性的两栖动物，和强大的5-hmC富集在神经元细胞之间的两栖动物和哺乳动物是保守的。此外，我们确定了5-hmC富集的细胞群分布在两栖动物的皮肤和结缔组织中的马赛克的方式。我们的研究结果表明，免疫化学不仅可以成功地用于空间识别的细胞富集5-hmC，但也为半定量评估的水平，这种表观遗传修饰在不同组织的单细胞。
5-Hydroxymethyl-cytosine (5-hmC) is a form of modified cytosine, which has recently attracted a considerable attention due to its potential role in transcriptional regulation. According to several reports 5-hydroxymethyl-cytosine distribution is tissue-specific in mammals. Thus, 5-hmC is enriched in embryonic cell populations and in adult neuronal tissue. Here, we describe a novel method of semi-quantitative immunohistochemical detection of 5-hmC and utilize it to assess the levels of this modification in amphibian tissues. We show that, similar to mammalian embryos, 5-hmC is enriched in axolotl tadpoles compared with adult tissues. Our data demonstrate that 5-hmC distribution is tissue-specific in amphibians, and that strong 5-hmC enrichment in neuronal cells is conserved between amphibians and mammals. In addition, we identify 5-hmC-enriched cell populations that are distributed in amphibian skin and connective tissue in a mosaic manner. Our results illustrate that immunochemistry can be successfully used not only for spatial identification of cells enriched with 5-hmC, but also for the semi-quantitative assessment of the levels of this epigenetic modification in single cells of different tissues.