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Effect of postmortem change on development of apoptosis in rats

死后变化对大鼠细胞凋亡发展的影响

基本信息

批准号：
08670474
负责人：
SAGISAKA Kaoru
金额：
$ 1.41万
依托单位：
Tohoku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08670474/
关键词：
apoptosis postmortem change stress Turnel TUNEL法

项目摘要

Terminal deoxynucleotidyl transferase (TdT) -mediated dUTP-digoxigenin nick end labeling (TUNEL) assay is useful to detect apoptotic cells in situ. We examined whether or not postmortem delay affects the development of apoptotic signals of cells in various organs by hematoxy lin-eosin (H-E) and TUNEL assay.Wistar Imamichi rats were radiated by X-ray and sacrificed six hours after radiation. The spleen, thymus, adrenal and testis were excised and kept in moist chamber at room temperature. Each tissue was fixed after different time intervals 0,6,12,24 hours, and paraffin-embedded sections were made. In no radiation group, a little of TUNEL positive cells were observed in the spleen, thymus and testis sections, but not in the adrenal. No increase in the number of apoptotic cells was observed with postmortem delay. In the radiation group, much increase in the number of TUNEL positive cells of which nuclei were clearly and deeply stained was observed in the spleen and thymus corresponding to the area where shrinking nuclei were observed in H-E section. In testis sections, there was a little increase in the number of positively stained cells, and no change was observed in H-E section. With postmortem delay, the margin of the TUNEL positive cells changed from clear to indistinct, and the positive area was spread around. Our results shows that it is difficult to distinguish apoptotic cell from postmortem change. It is possible, however, to detect TUNEL positive cell with postmortem changes as the spread TUNEL positive area.

末端脱氧核苷酸转移酶(TdT)介导的dUTP-地高辛缺口末端标记法(TUNEL)可用于原位检测细胞凋亡。采用苏木精-伊红(H-E)法和原位末端标记法(TUNEL)检测死后延迟是否影响不同器官细胞凋亡信号的发生。取脾、胸腺、肾上腺和睾丸，置于常温湿室保存。分别于术后0、6、12、24小时固定各组织，制作石蜡包埋切片。未照射组小鼠的脾、胸腺和睾丸切片中可见少量TUNEL阳性细胞，而肾上腺未见阳性细胞。随着死亡时间的延长，未观察到凋亡细胞的增加。照射组小鼠脾、胸腺与HE切片核缩小区相对应，TUNEL阳性细胞数明显增多，核染深染色明显。睾丸切片中阳性细胞数略有增加，而HE切片中未见明显变化。随着死后时间的延长，TUNEL阳性细胞的边缘由清晰变为模糊，阳性区域散在分布。我们的结果表明，很难区分死亡细胞和死亡细胞的变化。然而，有可能检测到死后发生变化的TUNEL阳性细胞作为扩散的TUNEL阳性区域。