碩士 / 國立臺灣海洋大學 / 食品科學系 / 100 / The cell and zinc levels in the blood of stressed carp are higher than those of resting carp, at the same time, the cell and zinc levels in head kidney of stressed carp are also higher than those of resting carp. Head kidney is the hematopoietic organ of common carp, the increases of cell level in the blood under stress, may be produced by head kidney. In order to understand how the cells in head kidney of carp proliferate, and the relationship between proliferation and zinc, market carp (stressed carp) and laboratory carp (resting carp) were used as materials. The cells in head kidney of 15 resting carps and 14 stressed carps were separated by Percoll density into 2 layers, layer 1 (ρ = 1.02) and 2 (ρ = 1.07), respectively, and the cell levels in different layers were measured. It was found that the cell levels in layer 1 of stressed carp is significantly higher than those in resting carp, being 532x106 cells/g tissue to 118x106 cells/g tissue. The cell levels in layer 2 were 746x106 cells/g tissue to 776x106 cells/g tissue, respectively, there is no significant difference between the stressed and resting carp. In layer 1, the main proliferated cells had a cell size of ~ 6 μm, for the stressed carp being 354x106 cells/g tissue, and for resting carp being 86x106 cells/g tissue, the difference was 4 times.
In light-microscopic observation of the Giemsa stain, most of the cells in layer 1 were round in size about 6 μm. They have thin layer cytoplasm and a round nucleus, it is very similar to hematopoietic stem cell/progenitor. The immunofluorescent stain with an antibody against the 43-kDa zinc-binding protein indicates there is more immunofluorescent response in the cells in layer 1 of the stressed carp. Ther results of staining with o-Dianisidine indicate there are more RBC in layer 2 of stressed carp.
The cells of common carp head kidney were serial subcultured with 4 types of medium: medium (DMEM/F12) only, medium + 10 % carp serum, medium + 10 % carp serum + 0.3mM ZnCl2, and medium + 0.3mM ZnCl2. At different time intervals, the cultured cells were separated into 2 layers by Percoll density, layer 1 (ρ = 1.02) and layer 2 (ρ = 1.07), respectivey; and the cell level was measured. It was found that the group of medium + 10% carp serum + 0.3mM ZnCl2 has the best growth. Zinc induced the cells in carp head kidney to proliferate, but needed the supplementation of carp serum.
In order to know the growth curve of the cell of carp head kidney under supplementation with or without zinc, the cells of carp head kidney were cultured with carp serum and zinc or carp serum only from h0 to h96. The cultured cells were then separated by Percoll density. The results indicate the cell levels in the group supplemented with carp serum and zinc of layer 1 at h0 was 3.60x106 cells/ml and at h96, 12.5 x106 cells/ml. The population doubling time of cells in layer 1 supplementaed carp serum and zinc was only 6 h, compared to that supplemented with serum only being 19 h. The layer 1 cells after being cultured for 3 days with zinc and carp serum were staind with an antibody against the 43-kDa zinc-binding protein. The results indicate that the 6 μm cells have immunofluorescent response. Besides, there are more RBC cells in layer 2. Zinc not only induce layer 1 cells to proliferate, but may also promote cell to differentiate to RBC.
When carps are under stress, zinc is released from its digestive tract tissue to blood, then transported to head kidney. Zinc is possibly bound to transferrin in blood. In the head kidney, zinc-transferrin complex may be bound to transferrin receptor on hematopoietic stem cell/progenitor cells of the head kidney. This may further the cells proliferate to become RBC to help the carps to overcome stress.
| Identifer | oai:union.ndltd.org:TW/100NTOU5253047 |
| Date | January 2012 |
| Creators | Szu-Wei Fang, 方思幃 |
| Contributors | Sen-Shyong Jeng, 鄭森雄 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 102 |
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