Transgenic mice with green fluorescent protein-labeled  pancreatic β-cells

Hara, Manami; Wang, Xiaoyu; Kawamura, Toshihiko; Bindokas, Vytas P.; Dizon, Restituto F.; Alcoser, Sergio Y.; Magnuson, Mark A.; Bell, Graeme I.

doi:10.1152/ajpendo.00321.2002

Cited by 285 publications

(304 citation statements)

References 25 publications

Supporting

Mentioning

294

Contrasting

Unclassified

Order By: Relevance

“…Wild-type (C57BL/6JJcl, CLEA Japan, Tokyo, Japan) mice, ROSA (B6.129S7-Gt(ROSA)26Sor/J, The Jackson Laboratory, Bar Harbor, ME) mice, MIP-GFP mice (9) and GFP Tg mice (C57BL/6-Tg(UBC-GFP) 30 Scha, The Jackson Laboratory) were used. Diabetes was induced by i.p.…”

Section: Methodsmentioning

confidence: 99%

Fusion of proinsulin-producing bone marrow-derived cells with hepatocytes in diabetes

Fujimiya

Kojima

Ichinose³

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

We previously reported that diabetes in mice is associated with the appearance of proinsulin-producing (Proins-P) cells in the liver. It was unclear, however, whether these Proins-P bone marrowderived cells (BMDC) merely transit through the liver or undergo fusion with hepatocytes, normally an extremely rare event. In this study, we found that, in diabetes, BMDC in the liver produce not only Proins but also TNF-␣, suggesting that diabetes reprograms gene expression in BMDC, turning on ''inappropriate'' genes. Bone marrow transplantation using genetically marked donor and recipient mice showed that fusion occurs between Proins-P BMDC and hepatocytes. Cell fusion is further supported by the presence of the Y chromosome in Proins-P cells in female mice that received male bone marrow transplantation cells. Morphologically, Proins-P fusion cells are albumin-producing hepatocytes that constitute Ϸ2.5% of the liver section area 5 months after diabetes induction. An extensive search failed to reveal any fusion cells in nondiabetic mice. Thus, diabetes causes fusion between Proins-P BMDC and hepatocytes in vivo, an observation that has implications for the pathophysiology of diabetes as well as the fundamental biology of heterotypic cell fusion.cell fusion ͉ diabetes mellitus ͉ diabetic complications ͉ liver R ecent reports from different laboratories document that bone marrow-derived cells (BMDC) can fuse with hepatocytes in vivo, and such fusion events can contribute to liver regeneration (1-4). However, with the exception of unique genetic models, e.g., fumarylacetoacetate hydrolase (Fah)-deficient mice (1, 2), in which wild-type BMDC confer a marked growth advantage, BMDC-hepatocyte fusion appears to be extremely rare. In a review article, Thorgeirsson and Grisham (5) estimated that transplanted BMDC generate hepatocyte-like cells in the liver at a frequency of Յ10 O4 . In fact, the yield of presumed BMDC-hepatocyte fusion cells ranges from 0 to Ͻ0.05% in Ͼ80% of reports published before August 2005 (5). Apart from the rarity of the event, there is little information on whether BMDC-hepatocyte cell fusion is regulated by physiological body functions, and whether it is affected by pathological conditions other than the extreme example of Fah-deficient mice that received bone marrow transplantation (BMT) from Fahexpressing mice.Our laboratory recently reported the appearance of proinsulin-producing (Proins-P) cells in the bone marrow of diabetic rats and mice (6). The Proins-P BMDC migrate out of the bone marrow and populate different organs and tissues, including liver and fat. Because the phenomenon occurs in both type 1 and type 2 models, and can be duplicated by glucose injections in mice, hyperglycemia appears to trigger its occurrence. This interpretation is supported by the observation that Ϸ50% of unfractionated BMDC start transcribing insulin within days of incubation in high-glucose medium (7).Diabetes mellitus is a prevalent disease that affects Ϸ7% of the U.S. population and a rapidly growing proportion o...

show abstract

Section: Methodsmentioning

confidence: 99%

Fusion of proinsulin-producing bone marrow-derived cells with hepatocytes in diabetes

Fujimiya

Kojima

Ichinose³

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…MIP-GFP transgenic mice 11 were used to isolate primary β-cells for in vitro studies. These mice express green fluorescent protein (GFP) in pancreatic β-cells under the control of the mouse insulin I promoter (MIP).…”

Section: Animalsmentioning

confidence: 99%

Glial Cell Line-Derived Neurotrophic Factor Increases β-Cell Mass and Improves Glucose Tolerance

et al. 2008

Self Cite

View full text Add to dashboard Cite

show abstract

“…The rat (Dahl et al 1996;Hanahan 1985;Vasavada et al 1996), mouse (Hara et al 2003), and human (Hotta et al 1998;Krakowski et al 1999) insulin promoters have been frequently used to direct expression of oncogenes, hormones, growth factors, transcription factors, reporter genes, and more recently of the enzyme Cre recombinase (Ahlgren et al 1998;Dor et al 2004;Gannon et al 2000;Postic et al 1999) specifically to pancreatic -cells in transgenic mice. A major caveat of the rat insulin promoter, however, is the reported ectopic expression in certain areas of the brain, potentially resulting in phenotypes in both -cells and neural cells (Gannon et al 2000;Martin et al 2003).…”

Section: Introductionmentioning

confidence: 99%

Specific transgene expression in mouse pancreatic β-cells under the control of the porcine insulin promoter

Grzech¹,

Dahlhoff²,

Herbach³

et al. 2010

Molecular and Cellular Endocrinology

View full text Add to dashboard Cite

The availability of regulatory sequences directing tissue-specific expression of transgenes in genetically modified mice and large animals is a prerequisite for the development of adequate models for human diseases. The rat insulin 2 gene (Ins2) promoter, widely used to achieve transgene expression in pancreatic -cells of mice, also directs expression to extrapancreatic tissues and performs poorly in isolated pancreatic islets of human, mouse, and pig. To evaluate whether the full 5' untranslated region (UTR) of the porcine insulin gene (INS) confers robust and specific expression in -cells we generated an expression cassette containing 1500 bp of the porcine INS 5' UTR and the 3' UTR of the bovine growth hormone gene (GH). The cassette was designed to allow easy exchange of the sequences to be expressed and easy removal of the vector backbone from the expression cassette. To evaluate the properties of the cassette, we initially cloned a cDNA encoding human betacellulin, a growth factor known to affect structural and functional parameters of -cells. After confirming the functionality and specificity of the construct in vitro, transgenic mouse lines were generated by pronuclear DNA microinjection. Using RT-PCR, immunohistochemistry and immunofluorescence, we show that transgenic mice expressed human betacellulin exclusively in -cells. Confirming the proposed insulinotropic effect of betacellulin, transgenic mice showed improved glucose tolerance. We conclude that the newly designed expression cassette containing 1.500 bp of the porcine insulin promoter 5' UTR confers robust and specific transgene expression to -cells in vitro and in transgenic mice.

show abstract

Transgenic mice with green fluorescent protein-labeled pancreatic β-cells

Cited by 285 publications

References 25 publications

Fusion of proinsulin-producing bone marrow-derived cells with hepatocytes in diabetes

Fusion of proinsulin-producing bone marrow-derived cells with hepatocytes in diabetes

Glial Cell Line-Derived Neurotrophic Factor Increases β-Cell Mass and Improves Glucose Tolerance

Specific transgene expression in mouse pancreatic β-cells under the control of the porcine insulin promoter

Contact Info

Product

Resources

About