Transgenic pigs with pancreas-specific expression of green fluorescent protein.
Transgenic pigs with pancreas-specific expression of green fluorescent protein.
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DOI：
10.1262/jrd.2014-006
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Nagashima H
中科院分区：
文献类型：
--
作者：
Matsunari H;Kobayashi T;Watanabe M;Umeyama K;Nakano K;Kanai T;Matsuda T;Nagaya M;Hara M;Nakauchi H;Nagashima H
由于胰腺的发育和再生在糖尿病等胰腺疾病中的作用，胰腺的发育和再生引起了人们极大的兴趣。在这里，我们试图开发一种可以追踪胰腺细胞谱系的大型动物模型。将胰腺和十二指肠同源盒-1 (Pdx1) 基因启动子与绿色荧光蛋白 Venus 缀合，并通过胞浆内精子注射介导的基因转移导入 370 个体外成熟的猪卵母细胞。这些卵母细胞被转移到四头受体母猪体内，所有母猪都怀孕了。三头后备母猪在妊娠 47-65 天时被处死，第四头被允许产仔。获得的 16 个胎儿中有 7 个是转基因 (Tg) 并表现出胰腺特异性的绿色荧光。第四只接受母猪产下一窝六头仔猪，其中两头是 Tg。创始人Tg后代正常成熟并产生健康的第一代（G1）后代。对四只 27 日龄 G1 Tg 仔猪的断奶后尸检证实了胰腺特异性 Venus 表达。胰腺组织的免疫染色表明转基因在β细胞中表达。将来自 Tg 猪的胰岛移植到 NOD/SCID 小鼠的肾囊下，并在移植后 1 个月内表达荧光。 Tg G1猪发育正常，血糖水平在正常范围内。性成熟前后胰岛素水平及其他血液生化参数均在正常范围内，表明胰腺功能正常。我们得出结论，Pdx1-Venus Tg 猪代表了适合胰腺发育/再生和糖尿病研究的大型动物模型。
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.
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DOI：
10.1089/clo.2008.0024
发表时间：
2008-09-01
期刊：
CLONING AND STEM CELLS
影响因子：
--
作者：
Matsunari, Hitomi;Onodera, Masafumi;Nagashima, Hiroshi
通讯作者：
Nagashima, Hiroshi
影响因子：
46.9
作者：
Nagai, T;Ibata, K;Miyawaki, A
通讯作者：
Miyawaki, A
影响因子：
64.8
作者：
JONSSON, J;CARLSSON, L;EDLUND, H
通讯作者：
EDLUND, H
影响因子：
64.8
作者：
HAMMER, RE;PURSEL, VG;BRINSTER, RL
通讯作者：
BRINSTER, RL
DOI：
10.1152/ajpendo.00321.2002
发表时间：
2003-01-01
影响因子：
5.1
作者：
Hara, M;Wang, XY;Bell, GI
通讯作者：
Bell, GI