食道為廣鹽性虱目魚的滲透壓調節器官

陳建宏

doi:10.6845/NCHU.2013.00788

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食道為廣鹽性虱目魚的滲透壓調節器官

Esophagus as an osmoregulatory organ of the euryhaline milkfish (Chanos chanos)

陳建宏 , Masters　　Advisor：李宗翰　　

繁體中文 DOI： 10.6845/NCHU.2013.00788

虱目魚；食道；滲透壓； milkfish ； esophagus ； osmolarity

Abstract │ Reference (76) │ Altmetrics

Abstract 〈TOP〉

硬骨魚類能藉由鰓、腎臟、腸三種主要的滲透壓調節器官，調節體內的離子與滲透壓平衡。此外在海水馴養的日本鰻(Anguilla japonica)和美洲擬鰈(Pseudopleuronectes americanus)已經證實食道為滲透壓調節器官，能將喝入的海水進行去鹽作用(desalination)，其管壁的表皮細胞具調節鹽類通透性的能力。本論文從細胞組織的巨觀到微觀進行比較，並結合生理實驗，探討廣鹽性硬骨魚類虱目魚(Chanos chanos)食道參與滲透壓調節的功能。首先利用解剖學的方式觀察虱目魚食道的內部型態，結果發現食道前段顏色較深且具有螺旋狀的皺褶，而後段則是透明且呈現縱向的皺褶。食道的組織切片化學染色顯示，馴養於淡水和海水的虱目魚食道管腔表皮細胞型態無顯著變化，但是淡水虱目魚食道前段與後段肌肉層的切面高度均較海水個體顯著較厚。藉由飲水實驗發現海水個體相較於淡水個體，其食道後段因吞嚥含有亞甲基藍的海水而被深染，此結果驗證馴養在海水中的虱目魚具有喝海水以補充水分的行為。進一步建立食道離體的生理實驗分析其離子通透性，結果顯示海水虱目魚的食道比淡水個體的食道具有較好的離子通透性。離子通透性的差異意味著其表皮細胞具有不同的離子通道參與其中。使用Na+, K+-ATPase (NKA)抑制劑ouabain探討NKA是否參與海水食道對離子的通透性，結果發現加入抑制劑的實驗組，其食道的通透性顯著低於控制組。偵測海水與淡水虱目魚的食道上之NKA α-subunit表現之差異，比較海水及淡水虱目魚食道的前段及後段管腔內的NKA的基因表現量、蛋白質含量、活性以及細胞(NKA immunoreactive cells)數量，結果顯示海水虱目魚食道前段的各項數值皆顯著高於淡水虱目魚組；而在食道後段各項數值則兩組間無明顯變化。據此推論食道前段NKA的調控參與個體喝入海水的去鹽作用。此外本論文偵測到二種氯離子通道蛋白在虱目魚食道前段上具有環境鹽度依賴性表現，CLC-5-like protein表現量在海水魚食道顯著高於淡水個體的食道。另外，NKCC2-like protein的表現量則是在淡水個體中較高。兩者表現位置都是在靠近管腔內螺旋狀皺褶的表皮細胞上。綜和以上實驗結果，虱目魚在不同鹽度環境中馴養時，食道會藉由細胞組織型態的改變以及調整滲透壓相關蛋白的表現，在海水去鹽作用上扮演重要的角色，而在淡水適應則參與離子吸收的功能。因此，食道亦為虱目魚的滲透壓調節的重要器官，參與消化道系統的離子調節以達到滲透壓平衡的生理狀態。

Parallel Abstract 〈TOP〉

To maintain homeostasis, marine teleosts drink seawater (SW) to supplement the internal water and excreted ions from gill, kidney and intestine. In esophagi of the eel (Anguilla japonica) and flounder (Pseudopleuronectes americanus), the epithelial cell exhibited desalination. In the present study, try to examine the osmoregulatory function of esophagus from marco to micro level in the milkfish (Chanos chanos). First, the internal morphologies indicated that there were many spiral valves in the anterior esophagus with deep color; while colorless posterior esophagus showed longitudinal wrinkles. These structures of esophagus are rarely described in other fishes. Histochemistry staining of esophageal sections revealed that no significant changes in epithelial cells between the freshwater (FW) and SW groups. Furthermore,  the muscular layer of FW group is thicker than SW group. Our results indicated that posterior esophagus of SW group was dyed by methylene blue. The drinking experiment was designed to investigate the concept that SW-acclimated milkfish drank SW. On the other hand, the system of isolated milkfish esophagus were measured esophageal ion permeability in vitro. The ion permeabilities in the SW groups were more than FW groups. Difference of esophageal permeability implied that there were ion channels involved in the ion transport. Ouabain-inhibitor was performed to determine the effects of NKA whether on the ion permeability in the esophagi of the SW fish. The permeability of esophagus in ouabain group was lower than control group (ouabain-free). Therefore, the NKA expression of anterior esophagus is the SW fish was higher than that of the FW group including gene level, protein level, NKA activity and NKA immunoreactive cell (NKA-IR) number. However, there was no significant difference in the posterior esophagus. In the present study, two kind of Cl- channels are identified in the anterior esophagus with salinity dependent. Expression of CLC-5-like protein in the SW fish was higher than in the FW group, expression of NKCC2-like protein in the FW group was higher than the SW group. Both of the ion transporters to the epithelial cells of spiral valves in the anterior esophagus. Taken together, esophagus was an osmoregulatory organ involved in ion regulation for homeostasis in euryhaline fish milkfish.

Reference ( 76 ) 〈TOP〉

Abaurrea-Equisoain, M. A., Ostos-Garrido, M. V. (1996). Cell types in the esophageal epithelium of Anguilla anguilla (Pisces, Teleostei). Cytochemical and ultrastructural characteristics. Micron. 27, 419-429.
連結：
Aguenaou, H., Boeuf, G., Colin, D. A. (1989). Na+ uptake through the brush border membranes of intestine from fresh water and sea-water adapted trout (Salmo gairdneri). J. Comp. Physiol. B. 159, 275-280.
連結：
Ando, M., Mukuda, T., Kozaka, T. (2003). Water metabolism in the eel acclimated to sea water: from mouth to intestine. Comp. Biochem. Physiol. B. 136, 621-633.
連結：
Bagarinao, T. (1994). Systematics, distribution, genetics and life history of milkfish, Chanos chanos. Env. Biol. Fishes. 39, 23-41.
連結：
Blanco, G., Mercer, R. W. (1998). Isozymes of the Na+-K+-ATPase: heterogeneity in structure, diversity in function. Am. J. Physiol. 275, 633-650.
連結：

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