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標題: 黑點青鱂魚(Oryzias dancena)之排氯機制:鰓蓋膜上富含粒線體細胞之研究及鰓上氯離子通道之分子表現
The chloride secretory mechanisms of the brackish medaka, Oryzias dancena: studies on the mitochondrion-rich cells of the opercular membrane and molecular expression of branchial chloride channel (cystic fibrosis transmembrane conductance regulator)
作者: 林上濤
Lin, Shang-Tao
關鍵字: Euryhaline teleost
Salinity tolerance
出版社: 生命科學系所
摘要: 本研究推論硬骨魚類的鰓蓋膜上普遍存在分布有許多富含粒線體細胞 (Mitochondria Rich, MR cells),此富含粒線體細胞據觀測顯示具有滲透壓調控的功能。研究的結果顯示,在此研究所用的模式物種“黑點青&;#40002;魚” brackish medaka (Oryzias dancena)的鰓蓋膜上所發現的富含粒線體細胞隨著鹽度環境變化而改變。改變的趨勢在細胞頂膜開口區的超微結構、細胞的大小與密度、Na+, K+, 2Cl&;#8722; cotransporter (NKCC)、Cystic fibrosis transmembrane conductance regulator (CFTR)等離子調控蛋白在MR細胞上的相對表現位置、鈉鉀氯離子共同通道蛋白(NKCC1a) 表現量,以及氯離子通透訊號的偵測等各種趨勢上都與先前在鰓的研究趨勢類似。另一方面,在將黑點青&;#40002;魚由海水環境直接轉移至淡水的實驗中發現,NKCC1a 蛋白在轉移中消失的速度較緩慢,並且在轉移後14天仍有部分NKCC1a蛋白未消失。然而,CFTR蛋白在轉移實驗中快速的消失。鰓上MR細胞的CFTR蛋白在一天內就完全消失。在鰓蓋膜中甚至發現CFTR蛋白在一小時內就完全消失了。此研究也完成了黑點青&;#40002;魚CFTR基因的序列全長,此序列與其他幾種廣鹽性模式魚種比對呈現高度的相似。研究也偵測了CFTR基因的mRNA表現量,並發現海水環境中鰓上的CFTR基因表現量顯著高於淡水環境魚鰓上的表現量。
The present study elucidated that MR cells were speculated to distribute generally in the opercular membrane of the teleosts and play the roles in osmoregulation. In our results of brackish medaka (Oryzias dancena), MR cells in the opercular membrane reacted to different salinity environments similarly to MR cells in gills reported in the previous studies, including the ultrastructures of apical region, cell size and density, NKCC1a protein expression, localization of ion regulatory protein, and the way of Cl- flux. High similarity between MR cells in these two tissues indicated that studies on MR cells of opercular membrane may apply to gills. On the other hand, the transferred experiments showed that the expression of CFTR and NKCC1a-like proteins changed when SW fish were directly exposed to FW. NKCC1a-like protein vanished in slower rate with small number remained over 14 days after transfer. However, CFTR protein vanished completely in 1 day. In the opercular membrane, CFTR even decayed immediately after transfer and vanished in 1 hour. In addition, the cftr gene of brackish medaka was cloned and showed high similarity to other model euryhaline teleosts. The CFTR mRNA expression in the gill was as expected significantly higher in the SW fish than FW fish.
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