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Transgenic Pigs with Pancreas-specific Expression of Green Fluorescent Protein

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...

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Autores principales: MATSUNARI, Hitomi, KOBAYASHI, Toshihiro, WATANABE, Masahito, UMEYAMA, Kazuhiro, NAKANO, Kazuaki, KANAI, Takahiro, MATSUDA, Taisuke, NAGAYA, Masaki, HARA, Manami, NAKAUCHI, Hiromitsu, NAGASHIMA, Hiroshi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Society for Reproduction and Development 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4085388/
https://www.ncbi.nlm.nih.gov/pubmed/24748398
http://dx.doi.org/10.1262/jrd.2014-006
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author MATSUNARI, Hitomi
KOBAYASHI, Toshihiro
WATANABE, Masahito
UMEYAMA, Kazuhiro
NAKANO, Kazuaki
KANAI, Takahiro
MATSUDA, Taisuke
NAGAYA, Masaki
HARA, Manami
NAKAUCHI, Hiromitsu
NAGASHIMA, Hiroshi
author_facet MATSUNARI, Hitomi
KOBAYASHI, Toshihiro
WATANABE, Masahito
UMEYAMA, Kazuhiro
NAKANO, Kazuaki
KANAI, Takahiro
MATSUDA, Taisuke
NAGAYA, Masaki
HARA, Manami
NAKAUCHI, Hiromitsu
NAGASHIMA, Hiroshi
author_sort MATSUNARI, Hitomi
collection PubMed
description 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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spelling pubmed-40853882014-07-08 Transgenic Pigs with Pancreas-specific Expression of Green Fluorescent Protein MATSUNARI, Hitomi KOBAYASHI, Toshihiro WATANABE, Masahito UMEYAMA, Kazuhiro NAKANO, Kazuaki KANAI, Takahiro MATSUDA, Taisuke NAGAYA, Masaki HARA, Manami NAKAUCHI, Hiromitsu NAGASHIMA, Hiroshi J Reprod Dev Original Article 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. The Society for Reproduction and Development 2014-04-21 2014-06 /pmc/articles/PMC4085388/ /pubmed/24748398 http://dx.doi.org/10.1262/jrd.2014-006 Text en ©2014 Society for Reproduction and Development http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives (by-nc-nd) License.
spellingShingle Original Article
MATSUNARI, Hitomi
KOBAYASHI, Toshihiro
WATANABE, Masahito
UMEYAMA, Kazuhiro
NAKANO, Kazuaki
KANAI, Takahiro
MATSUDA, Taisuke
NAGAYA, Masaki
HARA, Manami
NAKAUCHI, Hiromitsu
NAGASHIMA, Hiroshi
Transgenic Pigs with Pancreas-specific Expression of Green Fluorescent Protein
title Transgenic Pigs with Pancreas-specific Expression of Green Fluorescent Protein
title_full Transgenic Pigs with Pancreas-specific Expression of Green Fluorescent Protein
title_fullStr Transgenic Pigs with Pancreas-specific Expression of Green Fluorescent Protein
title_full_unstemmed Transgenic Pigs with Pancreas-specific Expression of Green Fluorescent Protein
title_short Transgenic Pigs with Pancreas-specific Expression of Green Fluorescent Protein
title_sort transgenic pigs with pancreas-specific expression of green fluorescent protein
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4085388/
https://www.ncbi.nlm.nih.gov/pubmed/24748398
http://dx.doi.org/10.1262/jrd.2014-006
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