title: Production of Cloned Miniature Pigs Expressing High Levels of Human Apolipoprotein(a) in Plasma.
abstract: High lipoprotein(a) [Lp(a)] levels are a major risk factor for the development of atherosclerosis. However, because apolipoprotein(a) [apo(a)], the unique component of Lp(a), is found only in primates and humans, the study of human Lp(a) has been hampered due to the lack of appropriate animal models. Using somatic cell nuclear transfer (SCNT) techniques, we produced transgenic miniature pigs expressing human apo(a) in the plasma. First, we placed the hemagglutinin (HA)-tagged cDNA of human apo(a) under the control of the β-actin promoter and cytomegalovirus enhancer, and then introduced this construct into kidney epithelial cells. Immunostaining of cells with anti-HA antibody allowed identification of cells stably expressing apo(a); one of the positive clones was used to provide donor cells for SCNT, yielding blastocysts that expressed apo(a). Immunohistochemical analysis of tissue sections and RT-PCR analysis of total RNA from organs of cloned piglet revealed that apo(a) is expressed in various tissues/organs including heart, liver, kidney, and intestine. More importantly, a transgenic line exhibited a high level (>400 mg/dL) of Lp(a) in plasma, and the transgenic apo(a) gene was transmitted to the offspring. Thus, we generated a human apo(a)-transgenic miniature pig that can be used as a model system to study advanced atherosclerosis related to human disease. The anatomical and physiological similarities between the swine and human cardiovascular systems will make this pig model a valuable source of information on the role of apo(a) in the formation of atherosclerosis, as well as the mechanisms underlying vascular health and disease.
authors.affiliation: Department of Biochemistry and Molecular Biology, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima, Japan.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Ozawa
authors.affiliation: Laboratory of Animal Reproduction, Faculty of Agriculture, Kagoshima University, Kagoshima, Japan.
authors.forename: Takehiro
authors.initials: T
authors.lastname: Himaki
authors.affiliation: Laboratory of Animal Reproduction, Faculty of Agriculture, Kagoshima University, Kagoshima, Japan.
authors.forename: Shoji
authors.initials: S
authors.lastname: Ookutsu
authors.affiliation: Laboratory of Animal Reproduction, Faculty of Agriculture, Kagoshima University, Kagoshima, Japan.
authors.forename: Yamato
authors.initials: Y
authors.lastname: Mizobe
authors.affiliation: Laboratory of Animal Reproduction, Faculty of Agriculture, Kagoshima University, Kagoshima, Japan.
authors.forename: Junki
authors.initials: J
authors.lastname: Ogawa
authors.affiliation: Laboratory of Animal Reproduction, Faculty of Agriculture, Kagoshima University, Kagoshima, Japan.
authors.forename: Kazuchika
authors.initials: K
authors.lastname: Miyoshi
authors.affiliation: Laboratory of Veterinary Clinical Pathology, Joint Faculty of Veterinary Medicine, Kagoshima University, Kagoshima, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Yabuki
authors.affiliation: Department of Molecular Pathology, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, Yamanashi, Japan.
authors.forename: Jianglin
authors.initials: J
authors.lastname: Fan
authors.affiliation: Laboratory of Animal Reproduction, Faculty of Agriculture, Kagoshima University, Kagoshima, Japan.
authors.forename: Mitsutoshi
authors.initials: M
authors.lastname: Yoshida
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics / Q000378:metabolism*; D053241:Apoprotein(a) / Q000096:biosynthesis* / Q000235:genetics; D050197:Atherosclerosis / Q000235:genetics / Q000378:metabolism; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism; D019976:Cloning, Organism; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D004195:Disease Models, Animal; D005260:Female; D006388:Hemagglutinins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008297:Male; D009928:Organ Specificity / Q000235:genetics; D013552:Swine / Q000235:genetics / Q000378:metabolism; D013556:Swine, Miniature / Q000235:genetics / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; D006388:Hemagglutinins; D053241:Apoprotein(a)