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研究生: 彭彥松
Peng, Yan-Sung
論文名稱: 前列腺素E2在斑馬魚酸鹼調節之功能
Function of PGE2 in zebrafish acid-base regulation
指導教授: 林豊益
Lin, Li-Yih
黃鵬鵬
Hwang, Pung-Pung
學位類別: 碩士
Master
系所名稱: 生命科學系
Department of Life Science
論文出版年: 2018
畢業學年度: 106
語文別: 英文
論文頁數: 55
中文關鍵詞: 斑馬魚前列腺素E2酸鹼平衡
英文關鍵詞: zebrafish, prostaglandin E2, acid-base regulation
DOI URL: http://doi.org/10.6345/THE.NTNU.SLS.015.2018.D01
論文種類: 學術論文
相關次數: 點閱:97下載:0
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  • 前列腺素E2(Prostaglandin E2)對於魚類酸鹼平衡的調節仍是未知的。PGE2可透過四型受體(EP),進而影響不同的生理功能。哺乳類主要利用腎臟維持酸鹼平衡,曾有研究發現PGE2參與兔子集尿管的排酸機制,而兩生類也需要透過表皮組織進行酸鹼平衡,排酸能力會受到不同的PGE2濃度而有所改變。然而,生活在水中的魚類,所需要面對環境改變的壓力比起前兩者更大,為了解PGE2與魚類酸鹼調節之間的關係,本研究將斑馬魚進行七天酸處理後發現鰓上PGE2相關的基因表現量上升。此外,將PGE2合成酵素弱化後,排酸能力、HR細胞數量以及排酸相關蛋白質的基因表現量上升。而將受體的基因ptger1a或ptger1b弱化後,在酸性環境下,可能藉由降低碳酸酐酶(ca2)基因表現量進而導致排酸能力無法上升。由此可知,PGE2可能參與斑馬魚的排酸機制,且在正常環境中的運作機制與酸性環境可能不同。

    How PGE2 regulate the acid-base regulation mechanism in fishes is unclear so far. It has been known that PGE2 regulates physiological function via four EP receptors (EP). One of the organs that could regulate acid-base balance in mammalian is the kidney. Previous study found that PGE2 inhibits proton secretion in medullary collecting duct of rabbits. On the other hand, in amphibians, PGE2 maintains the acid-base balance by utilizing epithelium and their proton secretion capability is dependent on the concentration of PGE2. However, Fishes, living in water, have to face stronger stress of environmental pH changes than previous species, especially in fresh water. In order to understand how does PGE2 affect proton secretion in fishes, we use zebrafish as a model animal. The acidic environmental acclimation of adult zebrafish found the up-regulation of PGE2 related genes in the gill. Additionally, knockdown PGE2 synthase (ptges) increases proton secretion ability, H+-ATPase rich (HR) cell number, and the expression of proton secretion related transporter genes were also up-regulated in zebrafish embryos. Furthermore, knockdown ptger1a or ptger1b showed lower proton secretion in acidic environment, it might caused by the down-regulation of ca2. Taken all together, PGE2 might involves in the machinery of acid-base regulation, however, it probably has different regulatory mechanisms between normal and acidic environment in zebrafish.

    Table of Contents 中文摘要 4 Abstract 5 Inoduction 6 Prostaglandins structure and receptors 6 Function of PGE2 in mammalian kidney 6 Function of PGs in acid-base balance in amphibian 7 Physiological function of PGE2 in fish 8 Zebrafish as a model to study acid-base homeostasis mechanism 9 Purpose 10 Materials and Methods 12 Experimental animals 12 Low pH acclimated experiment 12 Total RNA extraction 12 Reverse-transcription (RT) 13 Real-time quantitative PCR analysis (Q-PCR) 13 Whole-amount in situ hybridization 14 Morpholino oligonucleotides (MO) design and injection 15 H+-selective electrode technique 15 Measurement of whole body H+ efflux 16 Whole-mount immunocytochemistry and cell counting 17 Western blot analysis 18 Statistical analysis 18 Result 20 The effects of environmental pH to PGE2 related genes in zebrafish 20 Localization of ptges, ptger1a and ptger1b in zebrafish gill and 3 dpf embryos 20 The effects of ptges, ptger1a and ptger1b knockdown on 3 dpf embryos 21 The effects of ptges knockdown on the mRNA expression of transporters and the number of HR cells 22 The effects of ptger1a knockdown on the mRNA expression of transporters and the number of HR cells 23 The effects of ptger1b knockdown on the mRNA expression of transporters and the number of HR cells 24 Discussion 26 PGE2 might be involved in acid-base regulation in zebrafish 26 The protein of ptger1a and ptger1b might affect H+ secretion by regulating CA2 via cyclic AMP(cAMP) 26 The ptger1b might have effects on cell density 27 The up-regulation of nhe3b might be a compensation of down-regulation of rhcg1 in ptger1b morphant embryos 28 The role of PGE2 in zebrafish 29 Conclusion and Perspective 30 References 31 Table and figures 36

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