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Molecular Diagnosis of Genic Abnormalities using Human Tissue Materials

使用人体组织材料进行基因异常的分子诊断

基本信息

批准号：
07557326
负责人：
SHIOTA Kohei
金额：
$ 0.64万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07557326/
关键词：
Fluorescent in situ hybridization(FISH)DNA probes chromosome aneuploidye human embryos and fetuses fixed tissue retrospective diagnosis XO Turner fetus In situ hybrization FISH 性染色体特異的遺伝子性別判定性染色体異常ヒト胎児分子診断 FISH法性分化異常

项目摘要

1) A fluorescent in situ hybridization (FISH) study was carried out on paraffin sections of human embryonic and fetal tissues with two DNA probes, DXZ1 and DYZ1 (Oncor), for X and Y chromosome-specific DNA sequences, respectively. The paraffin blocks of the human embryonic and fetal tissues examined in the present study had been stored at room temperature for up to 5 years after fixation in 4% paraformaldehyde. All the 7 embryos and 5 fetuses examined were successfully sexed by FISH.The cells from 3 embryos and 4 fetuses were positive for a hybridization signal with each of the DXZ1 and DYZ1 probes and they were classified as male. The cells from the remaining 4 embryos and 1 fetus were positive for two identical hybridization signals with DXZ1 probe in a nucleus instead of the absence of the signal hybridized with DYZ1, indicating that their cells have two X chromosomes but no Y chromosomes. The FISH results for the 5 fetuses examined were consistent with their genital sex and/or gonadal histology. Thus, the FISH technique has been shown to visualize specific DNAs in situ on paraffin sections and to be useful to determine the sex of fixed embryos and fetuses retrospectively.2) A FISH study was carried out on paraffin sections of tissues from fetuses with 45, X karyotype which had been fixed in neutralized formalin. The results of the FISH with their tissues showed that the nucleus of each cell had one hybridization signal with the DXZ1 probe but was entirely negative for signals with the DYZ1 probe, indicating that the constitution of sex chromosomes of the fetuses was XO.The present results showed it feasible to retrospectively identify chromosome aneuploidies by FISH with chromosome-specific DNA probes if fixed tissues or paraffin blocks are available.

1)用DXZ 1和DYZ 1（Oncor）两种DNA探针分别对人胚胎和胎儿组织的石蜡切片进行荧光原位杂交（FISH）研究。本研究中检查的人胚胎和胎儿组织的石蜡块在4%多聚甲醛中固定后在室温下储存长达5年。7个胚胎和5个胎儿的性别均通过FISH鉴定，其中3个胚胎和4个胎儿的细胞与DXZ 1和DYZ 1探针杂交均为阳性，确定为雄性。其余4个胚胎和1个胎儿的细胞与DXZ 1探针在一个细胞核中出现两个相同的杂交信号，而不是没有与DYZ 1杂交的信号，表明它们的细胞具有两个X染色体，但没有Y染色体。检查的5只胎仔的FISH结果与其生殖器性别和/或性腺组织学一致。因此，FISH技术已被证明可以在石蜡切片上原位显示特定的DNA，并且可以用于确定固定的胚胎和胎儿的性别。2）对来自具有45，X核型的胎儿的组织的石蜡切片进行FISH研究，所述组织已被固定在中和的福尔马林中。组织FISH结果显示，DXZ 1探针在每个细胞核上均显示一个杂交信号，DYZ 1探针则完全阴性，表明胎儿性染色体组成为XO，说明在有固定组织或石蜡块的情况下，用染色体特异性DNA探针进行FISH技术回顾性鉴定染色体非整倍体是可行的。