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Bio-morphological studies of apoptotic with non-apoptotic dying cells in developing inner ear of mouse embryos.

小鼠胚胎发育内耳中凋亡与非凋亡死亡细胞的生物形态学研究。

基本信息

批准号：
15591839
负责人：
HARADA Tamotsu
金额：
$ 1.86万
依托单位：
Kawasaki Medical School
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

项目摘要

Dying cells studied the TdT-mediated dUTP nick end-labeling (TUNEL) method have been classified as ‘apoptotic' and ‘non-apoptotic' cells. In this study, in which 12-day-old mouse embryos were used because of a high frequency of ‘natural cell death' due to changing inner ear morphology, the percentages of ‘apoptotic' and ‘non-apoptotic' dying cells (ADC and NADC) among total dying cells in the inner ear were calculated.Observation of consecutive paraffin sections showed 90% of the dying inner ear cells to be ADC and 10% to by NADC in the control group. 70% of the inner ear dying cells were ADC and 30% were NADC in the cis-diammine-dichloroplatinum (CDDP) group ADC and NADC TUNEL positive dying cells in resin sections observed by light microscopy were examined again by transmission electron microscopy using a re-embedding procedure. ADC and NADC were then analyzed based on the classification of dying cells (type 1,2,3A and 3B) as described by Clarke. It was clear that ADC were the equivalent of type 1(apoptotic) dying cells and NADC were the equivalent of type 2(autophagic) dying cells. No type 3A or 3B cell death occurred. It was suggested that ADC essential to development of inner ear, and increased NADC might be defensive phenomenon against CDDP toxicity. We consider these finding to be important baselines for determining the process underlying abnormal development of the inner ear and its functional disorders such as hearing loss.

用TdT介导的dUTP缺口末端标记（TUNEL）方法研究的死亡细胞已被分类为“凋亡”和“非凋亡”细胞。在这项研究中，由于内耳形态的变化导致了高频率的“自然细胞死亡”，因此使用了12天大的小鼠胚胎，“凋亡”和“非凋亡”死亡细胞的百分比连续石蜡切片观察显示，90%的凋亡细胞为ADC，10%的凋亡细胞为NADC。对照组给予NADC。CDDP组染色细胞中70%为ADC，30%为NADC。光镜下观察到ADC和NADC染色阳性细胞，透射电镜下再包埋法检测染色细胞。然后基于如Clarke所述的垂死细胞的分类（1、2、3A和3B型）分析ADC和NADC。很明显，ADC相当于1型（凋亡）死亡细胞，NADC相当于2型（自噬）死亡细胞。未发生3A或3B型细胞死亡。提示ADC是内耳发育所必需的，NADC的增加可能是CDDP毒性的防御现象。我们认为这些发现是确定内耳异常发育及其功能障碍（如听力损失）的重要基线。