Inhibition of Hes1 activity in gall bladder epithelial cells promotes insulin expression and glucose responsiveness

R A Coad, J R Dutton, D Tosh, J Slack

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

The biliary system has a close developmental relationship with the pancreas, evidenced by the natural occurrence of small numbers of biliary-derived beta-cells in the biliary system and by the replacement of biliary epithelium with pancreatic tissue in mice lacking the transcription factor Hes1. In normal pancreatic development, Hes1 is known to repress endocrine cell formation. Here we show that glucose-responsive insulin secretion can be induced in biliary epithelial cells when activity of the transcription factor Hes1 is antagonised. We describe a new culture system for adult murine gall bladder epithelial cells (GBECs), free from fibroblast contamination. We show that Hes1 is expressed both in adult murine gall bladder and in cultured GBECs. We have created a new dominant negative Hes1 (Delta Hes1) by removal of the DNA-binding domain, and show that it antagonises Hes1 function in vivo. When Delta Hes1 is introduced into the GBEC it causes expression of insulin RNA and protein. Furthermore, it confers upon the cells the ability to secrete insulin following exposure to increased external glucose. GBEC cultures are induced to express a wider range of mature beta cell markers when co-transduced with Delta Hes1 and the pancreatic transcription factor Pdx1. Introduction of Delta Hes1 and Pdx1 can therefore initiate a partial respecification of phenotype from biliary epithelial cell towards the pancreatic beta cell.
Original languageEnglish
Pages (from-to)975-987
Number of pages13
JournalBiochemistry and Cell Biology - Biochimie Et Biologie Cellulaire
Volume87
Issue number6
DOIs
Publication statusPublished - 2009

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Urinary Bladder
Epithelial Cells
Insulin
Glucose
Transcription Factors
Biliary Tract
Endocrine Cells
Insulin-Secreting Cells
Fibroblasts
Cell culture
Pancreas
Contamination
Epithelium
Cell Culture Techniques
Cells
RNA
Tissue
Phenotype
DNA
Proteins

Cite this

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title = "Inhibition of Hes1 activity in gall bladder epithelial cells promotes insulin expression and glucose responsiveness",
abstract = "The biliary system has a close developmental relationship with the pancreas, evidenced by the natural occurrence of small numbers of biliary-derived beta-cells in the biliary system and by the replacement of biliary epithelium with pancreatic tissue in mice lacking the transcription factor Hes1. In normal pancreatic development, Hes1 is known to repress endocrine cell formation. Here we show that glucose-responsive insulin secretion can be induced in biliary epithelial cells when activity of the transcription factor Hes1 is antagonised. We describe a new culture system for adult murine gall bladder epithelial cells (GBECs), free from fibroblast contamination. We show that Hes1 is expressed both in adult murine gall bladder and in cultured GBECs. We have created a new dominant negative Hes1 (Delta Hes1) by removal of the DNA-binding domain, and show that it antagonises Hes1 function in vivo. When Delta Hes1 is introduced into the GBEC it causes expression of insulin RNA and protein. Furthermore, it confers upon the cells the ability to secrete insulin following exposure to increased external glucose. GBEC cultures are induced to express a wider range of mature beta cell markers when co-transduced with Delta Hes1 and the pancreatic transcription factor Pdx1. Introduction of Delta Hes1 and Pdx1 can therefore initiate a partial respecification of phenotype from biliary epithelial cell towards the pancreatic beta cell.",
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AU - Dutton, J R

AU - Tosh, D

AU - Slack, J

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AB - The biliary system has a close developmental relationship with the pancreas, evidenced by the natural occurrence of small numbers of biliary-derived beta-cells in the biliary system and by the replacement of biliary epithelium with pancreatic tissue in mice lacking the transcription factor Hes1. In normal pancreatic development, Hes1 is known to repress endocrine cell formation. Here we show that glucose-responsive insulin secretion can be induced in biliary epithelial cells when activity of the transcription factor Hes1 is antagonised. We describe a new culture system for adult murine gall bladder epithelial cells (GBECs), free from fibroblast contamination. We show that Hes1 is expressed both in adult murine gall bladder and in cultured GBECs. We have created a new dominant negative Hes1 (Delta Hes1) by removal of the DNA-binding domain, and show that it antagonises Hes1 function in vivo. When Delta Hes1 is introduced into the GBEC it causes expression of insulin RNA and protein. Furthermore, it confers upon the cells the ability to secrete insulin following exposure to increased external glucose. GBEC cultures are induced to express a wider range of mature beta cell markers when co-transduced with Delta Hes1 and the pancreatic transcription factor Pdx1. Introduction of Delta Hes1 and Pdx1 can therefore initiate a partial respecification of phenotype from biliary epithelial cell towards the pancreatic beta cell.

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