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人心脏房室结细胞的细胞荧光光度法核DNA测定

Cytofluorometric nuclear DNA determinations on the atrioventricular nodal cells in human hearts.

作者信息

Hayashi S, Takamatsu T, Fujita S

出版信息

Histochemistry. 1986;85(2):111-5. doi: 10.1007/BF00491756.

Abstract

In the human heart, it is well known that the polyploidization of working heart-muscle cells increases in proportion to increases in heart weight, but there has been no investigation of the process of polyploidization in the specialized heart-muscle cells of the cardiac conduction system which have a nerve-like function. In order to investigate the process of polyploidization in these cells, the nuclear DNA content of atrioventricular nodal cells was measured using cytofluorometry. Tissue samples taken from autopsied hearts without arrhythmias were embedded in paraffin blocks after Carnoy fixation. Blocks containing the atrioventricular conduction system were cut according to the serial sectioning method of Lev et al. The compact atrioventricular nodes were removed from thick paraffin sections (150 micron) under a stereomicroscope. The cells were then isolated by enzyme digestion and ultrasonic treatment. Smears of the isolated cells were double stained with azocarmin-G and acriflavine-Feulgen. Cytofluorometric DNA determinations of the DNA content of atrioventricular nodal cells were performed. Atrioventricular nodes were found to be composed of a large number of diploid cells and a small number of tetraploid cells. No octaploid cells were found. These findings reveal that the process of polyploidization in atrioventricular nodal cells is different from that found in working heart-muscle cells.

摘要

在人类心脏中,众所周知,工作心肌细胞的多倍体化与心脏重量的增加成比例增加,但对于具有神经样功能的心脏传导系统中的特殊心肌细胞的多倍体化过程,尚未有研究。为了研究这些细胞的多倍体化过程,使用细胞荧光测定法测量了房室结细胞的核DNA含量。取自无心律失常尸检心脏的组织样本在Carnoy固定后嵌入石蜡块中。根据Lev等人的连续切片方法,切取包含房室传导系统的石蜡块。在体视显微镜下从厚石蜡切片(150微米)中取出致密的房室结。然后通过酶消化和超声处理分离细胞。分离细胞的涂片用偶氮胭脂红-G和吖啶黄-福尔根进行双重染色。对房室结细胞的DNA含量进行细胞荧光测定。发现房室结由大量二倍体细胞和少量四倍体细胞组成。未发现八倍体细胞。这些发现表明,房室结细胞的多倍体化过程与工作心肌细胞中的不同。

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