碩士 / 國立臺灣海洋大學 / 食品科學系 / 101 / Abstract
Previous studies indicated that suspension culture of common carp head kidney cells with ZnCl2 and carp serum stimulated the proliferation of immature red blood cells at various stages of development. The active component in carp serum was identified to be transferrin. However, the mechanism how zinc and transferrin stimulate erythroid cell proliferation is still not clear. This study is attempted to understand how zinc and transferrin affect hematopoiesis in the common carp.
Effect of different zinc level on the growth of the carp head kidney cells was studied. We found that when the ZnCl2 level was below 0.1 mM, no cell growth of the immature red blood cells was observed. Maximal cell growth was observed when the ZnCl2 level was 0.3-0.6 mM (~200% of the control), and the half-maximal effect was observed at ~0.2 mM. Effect of supplementation of different carp serum % to the carp head kidney cells was also studied. Significant activation of cell growth was observed when 10 % of carp serum was added. If carp transferrin or human transferrin was replaced instead of carp serum, we found the most appropriate concentration was 0.05 μg/ml and 0.025-0.1 μg/ml, respectively. These results indicated that the proliferation of the cell only needed a small amount of transferrin. It seems that the growth rate of the immature red blood cells in carp head kidneys was regulated by the ZnCl2 level, tranferrin is not involved in the regulation.
In eukaryotes, transferrin is the major iron carrier and in the extracellular space of tissues. When the common carp head kidney cell suspension was cultured with FeCl2 or FeCl3 and carp serum, the results showed that ferric-transferrin complex had no effect on the red blood cell formation in the head kidney of the common carp. Besides, ferrin did not compete with zinc to bind transferrin.
In vivo, transferrin was isolated from the blood plasma of different degrees of
stressed common carp by Zn2+-IMAC (immobilized metal affinity
chromatography).The results showed that zinc-binding protein level in the plasma was
between 7.60 ± 1.74 to 11.2 ± 2.38 (mg / ml plasma). The separated zinc-binding
protein was analysed by electrophoresis. Coomassie blue-stained SDS-PAGE gels
showed that protein with approximate molecular mass of 70 kDa、65 kDa、60 kDa、
45 kDa were major components of the zinc-binding protein. The result indicated that
the transferrin concentration in the common carp blood was rather constant under
stress.
When the common carp is under stress, the zinc stored in the digestive tract
tissue of common carp was released from a 43-kDa zinc-binding protein located in its
digestive tract tissue. The released zinc interacts with transferrin in the blood plasma
and then transported to the head kidney where it is bound by the transferrin receptor
and triggers erythropoiesis. In this system, degrees of stress in common carp can
affect the zinc concentration in the head kidney, but not transferrin level in blood
plasma. In conclusion, transferrin serves as a transporter protein, the zinc was
transported into intracellular by transferrin. The growth rate of the immature red
blood cells in carp head kidneys was regulated by zinc level. Iron did not induce the
proliferation of immature red blood cells.
| Identifer | oai:union.ndltd.org:TW/101NTOU5253029 |
| Date | January 2013 |
| Creators | Chia-Ling Ho, 何家菱 |
| 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 | 47 |
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