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Immunotoxicological assay using fish neutrophil and toxic effects of the water pollutants

使用鱼中性粒细胞进行免疫毒理学测定和水污染物的毒性作用

基本信息

批准号：
15580160
负责人：
ENDO Makoto
金额：
$ 2.05万
依托单位：
Tokyo University of Marine Science and Technology (2004)東京水産大学 (2003)
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 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15580160/
关键词：
tilapia neutrophil estrogenic chemical swim bladder immunotoxity phagocytic activity

项目摘要

1.Study of tilapia neutrophil phagocytic activities after exposure to 17-beta estradiol (E2) : sensitivity in sexual differencesNeutrophil phagocytic activities were statically depressed after exposure to E2. Our appraisal method was not influenced by sex of the fish and this is a great advantage because it is not necessary to select the test fish depending on the their sex.2.Comparison of phagocytic activities between two cell populations isolated from the swim bladder and head kidney of tilapiaThe swim bladder neutrophil showed more consistent and repeatable results than the cells from the head kidney. This is because cell composition and maturation were more homogenous in the swim bladder. Isolation of cells from the swim bladder was easier than those from the head kidney.3.Expression of estrogen receptor (ER) α and βExpression of ER α and ER β were observed in neutrophil from both male and female fish using the in situ hybridization method. In the red blood cell, no signal was seen in both sexes.4.Protocol design to detect super oxide anion from the neutrophil1)Neutrophils (1.0 x 10^6 cells/50μl) were treated with the E2 (100 ppb) for 2 hrs.2)100μl of CLA(40μM), 300μl of PBS and 50μl of PMA(μg/ml) were added.3)The solution was mixed and the accumulating signal immediately was measured for 15 min. at 28℃.This protocol can quantify the intensity of chemiluminescence from exposed neutrophils to E2.

1. 17-β-雌二醇（E2）对罗非鱼中性粒细胞吞噬活性的影响：性别差异敏感性E2对中性粒细胞吞噬活性的抑制作用是静态的。2.罗非鱼鱼鳔和头肾两种细胞群吞噬活性的比较鱼鳔中性粒细胞比头肾中性粒细胞吞噬活性的测定结果具有更好的一致性和可重复性。这是因为鱼鳔中的细胞组成和成熟更均匀。雌激素受体（ER）α和β的表达用原位杂交方法观察了雌、雄鱼中性粒细胞中ER α和ER β的表达。在红细胞中，在两种性别中均未观察到信号。4.检测嗜中性粒细胞中超氧阴离子的方案设计1）中性粒细胞（1.0 × 10^6个细胞/50μl）用E2（100 ppb）处理2小时。2）100μl CLA（40μM），加入300μl PBS和50μl PMA（μg/ml）。3）混合溶液，立即在28℃下测量累积信号15分钟。该方案可以量化中性粒细胞对E2的化学发光。