Investigation of brain from infants with child abuse regarding to morphological change and epigenetic change such as DNA methylation.

本研究的目的是探讨虐待儿童的脑组织形态学改变和DNA甲基化等表观遗传学改变。采用尸检前CT、肉眼检查和显微镜下观察脑组织形态学变化。作为对照，大脑是从突然或意外死亡的情况下使用。受影响的人和对照组之间的大脑有任何显着的形态学差异。接下来，为了研究大脑中的DNA甲基化，进行了初步实验。我们证明了甲基化特异性PCR扩增可以使用从石蜡制备的DNA进行。用甲醛固定的脑组织包埋。然而，这是无效的，以评估DNA甲基化的程度。因此，我们需要一种方法来发现甲基化与婴儿压力显着相关的基因。因此，我们正在开发一种富集甲基化DNA的方法，该方法使用甲基化DNA与甲基化结合结构域1蛋白的沉淀，然后进行测序。将来，有可能通过该方法鉴定出甲基化与婴儿压力显着相关的基因。

The purpose of this study is to investigate brain from infants with child abuse regarding to morphological change and epigenetic change such as DNA methylation. Morphological change of the brain was investigated using postmortem CT prior to autopsy, naked eye examination and microscopic examination of the brain tissue. As control, brains were used from cases of sudden or unexpected death. There was any significant morphological difference in brains between the affected person and controls. Next, to investigate DNA methylation in brains, preliminary experiments were carried out. We demonstrated that methylation specific PCR amplification could be carried out using DNA prepared from the paraffin. embedded brain tissues which had been fixed by formaldehyde. However, it was invalid to evaluate the extent of DNA methylation. Therefore, we need a method to discover a gene of which methylation is significantly associated with stress in infants. Thus, we are developing a method for enrichment of methylated DNA using precipitation of methylated DNA with methylation binding domain 1 protein, followed by sequencing. In future, it might be possible to identify a gene of which methylation is significantly associated with stress in infants by the method.