Immunotoxicological assay using fish neutrophil and toxic effects of the water pollutants
Grant-in-Aid for Scientific Research (C)
|Allocation Type||Single-year Grants |
|Research Institution||Tokyo University of Marine Science and Technology (2004)|
ENDO Makoto Tokyo University of Marine Science and Technology, Graduate School of Marine Science and Technology, Professor, 海洋科学技術研究科, 教授 (80128355)
OKAMOTO Nobuaki Tokyo University of Marine Science and Technology, Faculty of Marine Science, Professor, 海洋科学部, 教授 (40114912)
MAITA Masashi Tokyo University of Marine Science and Technology, Graduate School of Marine Science and Technology, Associate Professor, 海洋科学技術研究科, 助教授 (60238839)
KATAGIRI Takayuki Tokyo University of Marine Science and Technology, Graduate School of Marine Science and Technology, Assistant Professor, 海洋科学技術研究科, 助手 (50361811)
|Project Period (FY)
2003 – 2004
Completed (Fiscal Year 2004)
|Budget Amount *help
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2004: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2003: ¥2,000,000 (Direct Cost: ¥2,000,000)
|Keywords||tilapia / neutrophil / estrogenic chemical / swim bladder / immunotoxity / phagocytic activity|
1.Study of tilapia neutrophil phagocytic activities after exposure to 17-beta estradiol (E2) : sensitivity in sexual differences
Neutrophil 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 tilapia
The 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 β
Expression 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 neutrophil
1)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.
Report (3 results)
Research Products (3 results)