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標題: 廣鹽性虱目魚馴養於不同鹽度環境中鰓上Na+/K+-ATPase α-subunit isoforms之表現變化
Switches of Na+/K+-ATPase α-subunit isoforms in gills of milkfish (Chanos chanos) acclimated to environments of different salinities
作者: 邱鈺惠
Chiu, Yu-Hui
關鍵字: Gill

Na+/K+-ATPase alpha-subunit
出版社: 生命科學系所
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摘要: 摘要 虱目魚(Chanos chanos)是一種廣鹽性硬骨魚類,無論在高張的海水環境或低張的淡水環境下皆可生存。先前的研究中指出,當虱目魚適應於不同的鹽度環境時,其鰓上Na+/K+-ATPase (NKA)活性有不同的變化,且其NKA對於鈉離子與鉀離子的親和力也不同。由此提出假說:NKA活性的變化及其對離子親和力的不同是來自於不同NKA α-subunit isoforms的表現。因此為進一步了解不同的NKA酵素特性是否與其α-subunit isoforms (α1,α2及α3)表現有關,本實驗將虱目魚馴養於高鹽海水(60‰)、海水(35‰)和淡水三種鹽度環境中,利用免疫轉漬法(immunoblot)從蛋白質含量來探討長期適應於上述三種環境,以及轉移鹽度環境(包括由海水轉移至淡水或高鹽海水)的短時間內,其鰓上NKA α-subunit isoforms表現情形。由實驗結果發現,長期馴養於海水的虱目魚,其鰓上NKA α1-isoform及NKA α3-isoform蛋白質含量顯著高於淡水及高鹽海水馴養之虱目魚。而在鹽度轉移的實驗部分,由海水轉移至高鹽海水時,NKA α1-isoform蛋白質含量在轉移48小時之前緩慢下降;當轉移96及168小時之後,則顯著降低,達未轉移前的四倍;NKA α2-isoform蛋白質含量在轉移期間,均無顯著變化,而NKA α3-isoform蛋白質含量,在轉移12小時後,即有顯著性的下降。另一方面,由海水轉移至淡水時,NKA α1-isoform蛋白質含量在起初的12小時有下降趨勢,之後隨即上升,至48小時有一高峰,當轉移96及168小時之後,再顯著性地下降,達未轉移前的三倍;NKA α2-isoform蛋白質含量則在轉移24小時後顯著性上升,之後下降回未轉移前相當;而NKA α3-isoform蛋白質含量,在轉移96及168小時後,即有顯著性的下降。綜合上述結果顯示,虱目魚鰓上NKA α1-isoform與NKA α3-isoform蛋白質含量上升,有助於適應在其原棲地-海水中,且當虱目魚長期適應或轉移至不同鹽度環境時,鰓上NKA α1-isoform與NKA α3-isoform的變化,有可能是導致此蛋白有不同的酵素特性的因素。
Abstract Milkfish (Chanos chanos) is an euryhaline teleost which is able to survive in environments with a broad range of salinities. Since previous studies revealed that upon salinity challenge milkfish exhibited adaptive changes in branchial NKA activity with different Na+ and K+ affinity, it was hypothesized that alteration in activity and ion-affinity was derived from changes of different isoforms of NKA α-subunit (i.e., the catalytic subunit). It is thus intriguing to compare the expression patterns of three major isoforms of NKA α-subunit (i.e., α1, α2, and α3) in gills of milkfish following salinity changes to realize their roles. In this study, milkfish were reared in seawater (SW, 35‰), fresh water (FW), or hypersaline water (HSW, 60‰) and the protein abundances of three NKA α-isoforms were analysed by immunoblotting. In acclimation experiments the milkfish were raised in SW, FW, or HSW for more than two weeks. The SW group exhibited significantly higher levels of NKA α1- and α3-subunit than the FW or HSW group. In addition, the time-course experiments for which milkfish were transferred from SW to FW or HSW and then sampled at 12, 24, 48, 96, and 168 h showed that after transfer from SW to HSW, NKA α1-isoform levels in gills declined gradually before 48 h and achieved a significant 4-folds decrease at 96 and 168 h, while NKA α3-isoform decreased significantly since 12 h post-transfer and no significant difference was found in NKA α2-isoform abundance. When fish were transferred from SW to FW, NKA α1-, α2-, α3-isoform levels decreased at 12 h post-transfer; α1-isoform increased gradually and peaked at 48 h while α2-isoform increased significantly at 24 h post-transfer. Then NKA α1- and α3-isoform levels achieved a significant decrease at 96 and 168 h following salinity transfer. Taken together, upregulation of gill NKA α1- and α3-isoforms was essential for the euryhaline milkfish to inhabit in SW, their primary natural habitat, and the changes of gill NKA α1- and α3-isoforms may lead to different Na+ or K+ affinities and fulfill some of the requirements for the subtly altered enzyme behaviors in gills of the marine euryhaline milkfish upon salinity challenge.
其他識別: U0005-2512200716573900
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