Department of Pathology, Institute of Basic Medical Sciences, University of Tsukuba, Tsukuba, 305-8575, Japan.
Cytotechnology. 2000 Jul;33(1-3):93-9. doi: 10.1023/A:1008115429679.
To study the functions of lipoprotein lipase (LPL) in lipid and lipoprotein metabolism and the relationship between LPL and atherosclerosis, we generated transgenic rabbits expressing the human LPL gene. A total of 4045 Japanese whiterabbit embryos were microinjected with a 3.8-kb SalI/HindIII fragment containing the chicken beta-actin promoter, human LPL cDNA and rabbit beta-globin with poly (A) signals, and then transplanted into 116 recipient rabbits. Of the 166 pups born, six pups were transgenic as confirmed by Southern blot analysis. ANorthern blot analysis revealed that human LPL was expressed by a number of tissues including the heart, kidney, adrenal gland and intestine. One transgenic rabbit showed up to 3-foldincreased LPL activity in post-heparin plasma compared to thatin nontransgenic rabbits. Human LPL expression in various tissues of transgenic rabbits was further elucidated by in situ hybridization and immunostaining. Since rabbits are superior to mice as a model of atherosclerosis, this transgenicrabbit model should provide a valuable tool for the study of LPL in lipid metabolism and atherosclerosis.
为了研究脂蛋白脂肪酶（LPL）在脂质和脂蛋白代谢中的功能以及 LPL 与动脉粥样硬化之间的关系，我们构建了表达人 LPL 基因的转基因兔。我们将一个含有鸡β-肌动蛋白启动子、人 LPL cDNA 和兔β-球蛋白多聚腺苷酸化信号的 3.8kb SalI/HindIII 片段显微注射到 4045 个日本白兔胚胎中，然后将这些胚胎移植到 116 只受体兔中。在 166 只出生的幼兔中，有 6 只经 Southern 印迹分析证实为转基因兔。Northern 印迹分析显示，人 LPL 在包括心脏、肾脏、肾上腺和肠道在内的多种组织中表达。与非转基因兔相比，一只转基因兔的肝素后血浆 LPL 活性增加了 3 倍。通过原位杂交和免疫染色进一步阐明了转基因兔各种组织中的人 LPL 表达。由于兔比鼠更适合作为动脉粥样硬化模型，因此这种转基因兔模型应该为研究 LPL 在脂质代谢和动脉粥样硬化中的作用提供有价值的工具。
