鹼性磷酸酶對斑馬魚腎臟發育及造血之影響

Effects of Alkaline Phosphatases on Zebrafish Kidney Development and Hematopoiesis

何佩瑾(Pei-Jin Ho)

指導教授 ： 謝錦源 ； 蔡淦仁

中山醫學大學/醫學科技學院/醫學檢驗暨生物技術學系碩士班/碩士(2010年)

https://doi.org/10.6834/CSMU.2010.00159

摘要

鹼性磷酸酶在人類中發現有組織非專一性鹼性磷酸酶、胎盤鹼性磷酸酶、生殖細胞鹼性磷酸酶、和腸道鹼性磷酸酶四種，在已知研究中發現組織非專一性鹼性磷酸酶除參與在造骨作用的礦化反應外，對於其他組織的鈣化作用（calcification）也有影響。目前斑馬魚中已知有組織非專一性鹼性磷酸酶的存在，在胚胎早期就有此基因之表現，在已知的實驗中發現組織非專一性鹼性磷酸酶表現在腎臟，而斑馬魚的腎臟為成熟斑馬魚造血的位置，其功能相似於哺乳類的骨髓。所以本研究是利用斑馬魚為模式動物來探討組織非專一性鹼性磷酸酶對斑馬魚腎臟發育及功能上影響。本研究是利用ALP moroholino（MO）顯微注射後發現胎胚發育延緩，眼睛與卵黃囊延伸區皆明顯變小變短。利用鹼性磷酸酶的受質觀測鹼性磷酸酶的表現，可以發現在腎臟的表現明顯減少。利用o-dianisidine 染色，可以觀察到紅血球的生成受到抑制。利用 neutral red 染色觀察巨噬細胞的生成，可以觀察到巨噬細胞在視網膜及尾部的地方皆有增加，也利用p53 MO將因 p53 上升造成巨噬細胞增加的可能性排除。總而言之，在腎臟的發育與腎臟的功能上，鹼性磷酸酶扮演重要角色。

關鍵字

鹼性磷酸酶腎臟 ； 造血

並列摘要

There are four isozymes of alkaline phosphatase (ALP) in the human, tissue nonspecific alkaline phosphatase (TNAP), placental alkaline phosphatase (PLAP), germ cell alkaline phosphatase (GCAP), and intestinal alkaline phosphatase (IAP). The recent studies showed that TNAP was not only involved in the mineralization, but also involved in calcification. From our previous study, it is known that TNAP expressed at early developmental stages of zebrafish, especially at kidney, the adult organ for hematopoiesis that is similar to the bone marrow of mammals. The aims of the study were to elucidate the effect of TNAP on zebrafish kidney development and function. After TNAP morpholino injection, the zebrafish embryogenesis was delay, the width and length of eyes and yolk extension were smaller and shorter than WT control, respectively. Besides, by ALP substrate staining, the expression of ALP was decreased. Using o-dianisidine staining, the hematopoiesis of RBC was decreased. By neutral red staining, the macrophages were increased in the retina and in the posterior of trunk. ALP MO was co-injected with p53 MO to exclusion the possibility that the increased number of macrophages were caused by the p53 over-expression. In conclusion that ALP plays very important roles in kidney development and function.

並列關鍵字

alkaline phosphatase ； kidney ； hematopoiesis

參考文獻

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