本論文研究包括選殖定序脂肪代謝相關基因脂蛋白脂解酶 (Lipoprotein lipase；LPL)和脂肪酸結合蛋白 (Fatty acid-binding proteins；FABPs)基因，並以Real-time PCR分析餵飼不同油脂處理的海鱺幼魚該二基因mRNA及上游調控PPARs基因之表現情形。不同油脂實驗飼料油脂含量15%，其中6% 為魚油，9％分別添加含n-3 HUFA為主的魚油 (FO)、含18:3 n-3為主的紫蘇油 (PE)、含18:2 n-6為主的紅花油 (SA)、含18:1 n-9為主的橄欖油(OL)或含16:0為主的棕櫚油 (PA)。本研究成功選殖海鱺LPL520個胺基酸全長序列、L-FABP 126個胺基酸全長序列和M-FABP 133個胺基酸全長序列。以這些序列進行RT-PCR與real time PCR，發現不同油脂飼料在海鱺各組織L-FABP和M-FABP mRNA的表現有顯著差異，相對的， LPL mRNA的表現除脂肪團外，在各組織間表現量則不受飼料油脂顯著影響。整體而言，飼料脂肪酸含量與LPL mRNA表現在肝臟與肌肉中的表現呈正相關，但在脂肪團中的表現卻呈負相關；FABPs mRNA的表現量也與飼料脂肪酸含量呈正相關。各脂肪酸中，C14:0、C20:1 n-9、EPA及DHA的含量與LPL及FABPs上游調控基因PPARγ mRNA的表現量呈正相關，也與脂肪團FABPs mRNA及肌肉LPL mRNA表現量呈正相關。因此，海鱺LPL、L-FABP及M-FABP mRNA的表現顯著受飼料脂肪酸種類及含量影響，而PPARs在其中的角色則相當複雜，並未清楚顯示。

The present study cloned successfully two lipid-metabolism genes, lipoprotein lipase (LPL) and fatty acid binding protein (FABPs) from cobia and studied the mRNA expressions of the two genes and their upstream gene PPARs when the cobia were fed diets containing 15% lipid. Among the lipids, 6% was fish oil and the remaining 9% were supplemented by fish oil (FO, rich in n-3 HUFA), perilla oil (PE, rich in 18:2 n-6), safflower oil (SA, rich in 18:2 n-6), olive oil (OL, rich in 18:1 n-9) or palm oil (PA, rich in 16:0). The whole sequences of LPL, liver-FABP (L-FABP) and muscle-FABP (M-FABP) encode 520, 126 and 133 amino acids, respectively. RT-PCR and real time PCR analyses based on these gene sequences show that the mRNA expressions of L-FABP and M-FABP in the tissue of the cobia were diet-specific. The mRNA expression of LPL, on the other hand, did not respond to the treatments, except in visceral fat depot. Linear regression analysis shows that the mRNA expression of LPL in the liver and muscle was positively (P<0.05) related to dietary fatty acids and ther concentration, but that in the visceral fat depot was negatively related. The mRNA expression of FABPs was also positively correlated with dietary fatty acid levels. Among all fatty acids, the levels of C14:0, C20:1 n-9, EPA and DHA were positively correlated with the mRNA expression of PPARγand also with FABPs mRNA expression in the visceral fat depot and LPL mRNA expression in the muscle. Thus, LPL, L-FABP and M-FABP mRNA expression of the cobia were highly influenced by the kind and amount of dietary fatty acids. The role of PPARs was not clearly demonstrated.

目錄
中文摘要 Ⅰ
英文摘要 Ⅱ
目錄 Ⅳ
表目錄 Ⅴ
圖目錄 Ⅶ
一、文獻整理 1
二、前言 16
三、材料與方法 18
四、結果 34
五、討論 93
六、結論 97
七、參考文獻 98

丁詩同、劉秉勳和許哲銘，2003。多不飽和脂肪酸調控脂質代謝基因表達之機制。科學農業51:160-168。
李佳芳，2005。點帶石斑魚（Epinephelus coioides）脂肪細胞補體相關蛋白30 kDa（ACRP30）基因之分子選殖及其對葡萄糖與脂質代謝之影響。國立台灣大學水產養殖學系碩士論文。
李愛杰，1998。水產動物營養與飼料學。
周本善，1997。吳郭魚脂質需求之探討。國立台灣大學水產食品科學研究所博士論文。
林佩貞，2008。飼料添加植物油脂對海鱺稚魚成長、脂肪狀態、代謝酵素活性、及脂肪酸組成的影響。國立中山大學海洋生物研究所碩士論文。
許珊菁，2006。鼠模式中高脂飲食、肥胖與脂質調控基因之表現。臺灣大學
微生物與生化學研究所博士論文。
陳威志，2006。以亞麻仁油和玉米油取代於油對海鱺幼魚成長、脂肪酸代謝及免疫反應的影響。國立中山大學海洋生物研究所碩士論文。
黃何興，2000。餌料油脂對海鱺幼魚成長與體組成的影響。國立中山大學海洋生物研究所碩士論文。
蔡美玲，2007。海鱺過氧化體增生活化受體(PPARs)基因表現對體脂肪與脂肪生
成的影響。國立中山大學海洋生物研究所博士班論文計劃書。

Albalat, A., A. Saera-Vila, E. Capilla, J. Gutierrez, J. Perez-Sanchez, and I. Navarro. 2007. Insulin regulation of lipoprotein lipase (LPL) activity and expression in gilthead sea bream (Sparus aurata). Comparative Biochemistry and Physiology Part B, Biochemistry and Molecular Biology 148(2): 151-159.
Ando, S., X. H. Xue, G. F. Tibbits, and N. H. Haunerland. 1998. Cloning and sequencing of complementary DNA for fatty acid binding protein from rainbow trout heart. Comparative Biochemistry and Physiology Part B, Biochemistry and Molecular Biology 119(1): 213-217.
Bell, J. G., J. McEvoy, D. R. Tocher, F. McGhee, P. J. Campbell, and J. R. Sargent. 2001. Replacement of fish oil with rapeseed oil in diets of Atlantic salmon (Salmo salar) affects tissue lipid compositions and hepatocyte fatty acid metabolism. The Journal of Nutrition 131(5): 1535-1543.
Bell, J. G., D. R. Tocher, R. J. Henderson, J. R. Dick, and V. O. Crampton. 2003. Altered fatty acid compositions in atlantic salmon (Salmo salar) fed diets containing linseed and rapeseed oils can be partially restored by a subsequent fish oil finishing diet. The Journal of Nutrition 133(9): 2793-2801.
Bernlohr, D. A., M. A. Simpson, A. V. Hertzel, and L. J. Banaszak. 1997. Intracellular lipid-binding proteins and their genes. Annual Review of Nutrition 17: 277-303.
Beynen, A. C., and M. B. Katan. 1985. Why do polyunsaturated fatty acids lower serum cholesterol? The American Journal of Clinical Nutrition 42(3): 560-563.
Bocos, C., M. Gottlicher, K. Gearing, C. Banner, E. Enmark, M. Teboul, A. Crickmore, and J. A. Gustafsson. 1995. Fatty acid activation of peroxisome proliferator-activated receptor (PPAR). The Journal of Steroid Biochemistry and Molecular Biology 53(1-6): 467-473.
Botham, K. M., X. Zheng, M. Napolitano, M. Avella, C. Cavallari, R. Rivabene, and E. Bravo. 2003. The effects of dietary n-3 polyunsaturated fatty acids delivered in chylomicron remnants on the transcription of genes regulating synthesis and secretion of very-low-density lipoprotein by the liver: modulation by cellular oxidative state. Experimental Biology and Medicine (Maywood.) 228(2): 143-151.
Cheng, W., L. Guo, Z. Zhang, H. M. Soo, C. Wen, W. Wu, and J. Peng. 2006. HNF factors form a network to regulate liver-enriched genes in zebrafish. Developmental Biology 294(2): 482-496.
Chmurzynska, A. 2006. The multigene family of fatty acid-binding proteins (FABPs): function, structure and polymorphism. Journal of Applied Genetics 47(1): 39-48.
Desvergne, B., and W. Wahli. 1999. Peroxisome proliferator-activated receptors: nuclear control of metabolism. Endocrine Reviews 20(5): 649-688.
Escher, P., and W. Wahli. 2000. Peroxisome proliferator-activated receptors: insight into multiple cellular functions. Mutation Research 448: 121-138.
Esteves, A., and R. Ehrlich. 2006. Invertebrate intracellular fatty acid binding proteins. Comparative Biochemistry and Physiology Part C: Toxicology and Pharmacology 142(3-4): 262-274.
Fielding, B. A. and K. N. Frayn. 1998. Lipoprotein lipase and the disposition of dietary fatty acids. The British Journal of Nutrition 80(6): 495-502.
Fauconneau, B., H. Alami-Durante, M. Laroche, J. Marcel, and D. Vallot. 1995. Growth and meat quality in carp. Aquaculture 129, 265-297.
Jordal, A. E., I. Hordvik, M. Pelsers, D. A. Bernlohr, and B. E. Torstensen. 2006. FABP3 and FABP10 in Atlantic salmon (Salmo salar L.)--general effects of dietary fatty acid composition and life cycle variations. Comparative Biochemistry and Physiology Part B, Biochemistry and Molecular Biology 145(2): 147-158.
Jump, D. B., S. D. Clarke, A. Thelen, M. Liimatta, B. Ren, and M. Badin. 1996. Dietary polyunsaturated fatty acid regulation of gene transcription. Progress in Lipid Research 35(3): 227-241.
Kendrick, J. S., and J. A. Higgins. 1999. Dietary fish oils inhibit early events in the assembly of very low density lipoproteins and target apoB for degradation within the rough endoplasmic reticulum of hamster hepatocytes. Journal of Lipid Research 40(3): 504-514.
Kleveland, E. J., B. Ruyter, A. Vegusdal, H. Sundvold, R. K. Berge, and T. Gjoen. 2006. Effects of 3-thia fatty acids on expression of some lipid related genes in Atlantic salmon (Salmo salar L.). Comparative Biochemistry and Physiology Part B, Biochemistry and Molecular Biology 145(2): 239-248.
Kondo, H., R. Misaki, L. Gelman, and S. Watabe. 2007. Ligand-dependent transcriptional activities of four torafugu pufferfish Takifugu rubripes peroxisome proliferator-activated receptors. General and Comparative Endocrinology 154(1-3): 120-127.
Liang, X. F., H. Y. Ogata, and H. Oku. 2002a. Effect of dietary fatty acids on lipoprotein lipase gene expression in the liver and visceral adipose tissue of fed and starved red sea bream Pagrus major. Comparative Biochemistry and Physiology Part A: Molecular and Integrative Physiology 132(4): 913-919.
Liang, X. F., H. Oku, and H. Y. Ogata. 2002b. The effects of feeding condition and dietary lipid level on lipoprotein lipase gene expression in liver and visceral adipose tissue of red sea bream Pagrus major. Comparative Biochemistry and Physiology Part A: Molecular and Integrative Physiology 131(2): 335-342.
Lindberg, A., and G. Olivecrona. 2002. Lipoprotein lipase from rainbow trout differs in several respects from the enzyme in mammals. Gene 292(1-2): 213-223.
Mead, J. R., S. A. Irvine, and D. P. Ramji. 2002. Lipoprotein lipase: structure, function, regulation, and role in disease. Journal of Molecular Medicine 80(12): 753-769.
Merkel, M., R. H. Eckel, and I. J. Goldberg. 2002. Lipoprotein lipase: genetics, lipid uptake, and regulation. Journal of Lipid Research 43(12): 1997-2006.
Montalto, M. B., and A. Bensadoun. 1993. Lipoprotein lipase synthesis and secretion: effects of concentration and type of fatty acids in adipocyte cell culture. Journal of Lipid Research 34(3): 397-407.
Mourente, G., J. R. Dick, J. G. Bell, and D. R. Tocher. 2005. Effect of partial substitution of dietary fish oil by vegetable oils on desaturation and β-oxidation of [1-14C] 18:3 n-3 (LNA) and [1-14C] 20:5 n-3 (EPA) in hepatocytes and enterocytes of European sea bass (Dicentrarchus labrax L.). Aquaculture 248: 173-186.
Murphy, M. C., A. Zampelas, S. M. Puddicombe, N. P. Furlonger, L. M. Morgan, and C. M. Williams. 1993. Pretranslational regulation of the expression of the lipoprotein lipase (EC 3.1.1.34) gene by dietary fatty acids in the rat. The British Journal of Nutrition 70(3): 727-736.
Niot, I., H. Poirier, and P. Besnard. 1997. Regulation of gene expression by fatty acids: special reference to fatty acid-binding protein (FABP). Biochimie 79(2-3): 129-133.
Oku, H., N. Koizumi, T. Okumura, T. Kobayashi, and T. Umino. 2006. Molecular characterization of lipoprotein lipase, hepatic lipase and pancreatic lipase genes: effects of fasting and refeeding on their gene expression in red sea bream Pagrus major. Comparative Biochemistry and Physiology Part B, Biochemistry and Molecular Biology 145(2): 168-178.
Oku, H., H. Y. Ogata, and X. F. Liang. 2002. Organization of the lipoprotein lipase gene of red sea bream Pagrus major. Comparative Biochemistry and Physiology Part B, Biochemistry and Molecular Biology 131(4): 775-785.
Owen, J. M., J. W. Adron, C. Middleton, and C. B. Cowey. 1975. Elongation and desaturation of dietary fatty acids in turbot Scophtalmus maximus L., and rainbow trout, Salmo gairdnerii rich. Lipids 10(9): 528-531.
Raclot, T., R. Groscolas, D. Langin, and P. Ferre. 1997. Site-specific regulation of gene expression by n-3 polyunsaturated fatty acids in rat white adipose tissues. Journal of Lipid Research 38(10): 1963-1972.
Richard, N., G. Mourente, S. Kaushik, and G. Corraze. 2006. Replacing dietary fish oil by vegetable oils has little effect on lipogenesis, lipid transport and tissue lipid uptake in rainbow trout (Oncorhynchus mykiss). Aquaculture 261: 1077-1087.
Saera-Vila, A., J. A. Calduch-Giner, P. Gomez-Requeni, F. Medale, S. Kaushik, and J. Perez-Sanchez. 2005. Molecular characterization of gilthead sea bream (Sparus aurata) lipoprotein lipase. Transcriptional regulation by season and nutritional condition in skeletal muscle and fat storage tissues. Comparative Biochemistry and Physiology Part B, Biochemistry and Molecular Biology 142(2): 224-232.
Schoonjans, K., B. Staels, and J. Auwerx. 1996. The peroxisome proliferator activated receptors (PPARs) and their effects on lipid metabolism and adipocyte differentiation. Biochimica et Biophysica Acta 1302(2): 93-109.
Scott, J. C., M. W. Kennedy, and D. P. McManus. 2000. Molecular and immunological characterisation of a polymorphic cytosolic fatty acid binding protein from the human blood fluke of humans, Schistosoma japonicum. Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression 1517(1): 53-62.
Sharma, M. K., E. M. Ovan-Wright, A. Degrave, C. Thisse, B. Thisse, and J. M. Wright. 2004. Sequence, linkage mapping and early developmental expression of the intestinal-type fatty acid-binding protein gene (fabp2) from zebrafish (Danio rerio). Comparative Biochemistry and Physiology Part B, Biochemistry and Molecular Biology 138(4): 391-398.
Spector, A. A., and M. A. Yorek. 1985. Membrane lipid composition and cellular function. Journal of Lipid Research 26(9): 1015-1035.
Storch, J., and A. E. A. Thumser. 2000. The fatty acid transport function of fatty acid-binding proteins. Biochimica et Biophysica Acta 1486: 28-44.
Tocher, D. R. 2003. Metabolism and functions of lipids and fatty acids in teleost fish. Reviews in Fisheries Science 11(2): 107-184.
Tocher, D. R., J. Carr, and J. R. Sargent. 1989. Polyunsaturated fatty acid metabolism in fish cells: differential metabolism of (n-3) and (n-6) series acids by cultured cells originating from a freshwater teleost fish and from a marine teleost fish. Comparative Biochemistry and Physiology B. Comparative Biochemistry 94(2): 367-374.
Tsfadia, Y., R. Friedman, J. Kadmon, A. Selzer, E. Nachliel, and M. Gutman. 2007. Molecular dynamics simulations of palmitate entry into the hydrophobic pocket of the fatty acid binding protein. FEBS Letters 581(6): 1243-1247.
Wilkinson, J., J. A. Higgins, C. Fitzsimmons, and D. E. Bowyer. 1998. Dietary fish oils modify the assembly of VLDL and expression of the LDL receptor in rabbit liver. Arteriosclerosis, Thrombosis, and Vascular Biology 18(9): 1490-1497.
Zheng, X., I. Seiliez, N. Hastings, D. R. Tocher, S. Panserat, C. A. Dickson, P. Bergot, and A. J. Teale. 2004. Characterization and comparison of fatty acyl Delta6 desaturase cDNAs from freshwater and marine teleost fish species. Comparative Biochemistry and Physiology Part B, Biochemistry and Molecular Biology 139(2): 269-279.
Zheng, X., B. E. Torstensen, D. R. Tocher, J. R. Dick, R. J. Henderson, and J. G. Bell. 2005. Environmental and dietary influences on highly unsaturated fatty acid biosynthesis and expression of fatty acyl desaturase and elongase genes in liver of Atlantic salmon (Salmo salar). Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids 1734(1): 13-24.