title: Transgenic pigs with pancreas-specific expression of green fluorescent protein.
abstract: The development and regeneration of the pancreas is of considerable interest because of the role of these processes in pancreatic diseases, such as diabetes. Here, we sought to develop a large animal model in which the pancreatic cell lineage could be tracked. The pancreatic and duodenal homeobox-1 (Pdx1) gene promoter was conjugated to Venus, a green fluorescent protein, and introduced into 370 in vitro-matured porcine oocytes by intracytoplasmic sperm injection-mediated gene transfer. These oocytes were transferred into four recipient gilts, all of which became pregnant. Three gilts were sacrificed at 47-65 days of gestation, and the fourth was allowed to farrow. Seven of 16 fetuses obtained were transgenic (Tg) and exhibited pancreas-specific green fluorescence. The fourth recipient gilt produced a litter of six piglets, two of which were Tg. The founder Tg offspring matured normally and produced healthy first-generation (G1) progeny. A postweaning autopsy of four 27-day-old G1 Tg piglets confirmed the pancreas-specific Venus expression. Immunostaining of the pancreatic tissue indicated the transgene was expressed in β-cells. Pancreatic islets from Tg pigs were transplanted under the renal capsules of NOD/SCID mice and expressed fluorescence up to one month after transplantation. Tg G1 pigs developed normally and had blood glucose levels within the normal range. Insulin levels before and after sexual maturity were within normal ranges, as were other blood biochemistry parameters, indicating that pancreatic function was normal. We conclude that Pdx1-Venus Tg pigs represent a large animal model suitable for research on pancreatic development/regeneration and diabetes.
authors.affiliation: Meiji University International Institute for Bio-Resource Research, Kawasaki 214-8571, Japan.
authors.forename: Hitomi
authors.initials: H
authors.lastname: Matsunari
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Kobayashi
authors.forename: Masahito
authors.initials: M
authors.lastname: Watanabe
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Umeyama
authors.forename: Kazuaki
authors.initials: K
authors.lastname: Nakano
authors.forename: Takahiro
authors.initials: T
authors.lastname: Kanai
authors.forename: Taisuke
authors.initials: T
authors.lastname: Matsuda
authors.forename: Masaki
authors.initials: M
authors.lastname: Nagaya
authors.forename: Manami
authors.initials: M
authors.lastname: Hara
authors.forename: Hiromitsu
authors.initials: H
authors.lastname: Nakauchi
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Nagashima
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D058948:Cell Tracking / Q000379:methods / Q000662:veterinary; D005260:Female; D018507:Gene Expression Regulation, Developmental; D018014:Gene Transfer Techniques / Q000662:veterinary; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D018398:Homeodomain Proteins / Q000235:genetics; D016381:Islets of Langerhans Transplantation / Q000379:methods / Q000662:veterinary; D008297:Male; D009928:Organ Specificity / Q000235:genetics; D010179:Pancreas / Q000196:embryology / Q000378:metabolism*; D011247:Pregnancy; D020554:Sperm Injections, Intracytoplasmic / Q000662:veterinary; D013552:Swine / Q000196:embryology / Q000235:genetics*; D015534:Trans-Activators / Q000235:genetics
chemical_list: D018398:Homeodomain Proteins; D015534:Trans-Activators; C084010:pancreatic and duodenal homeobox 1 protein; D049452:Green Fluorescent Proteins
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