title: Endocrine-committed progenitor cells retain their differentiation potential in the absence of neurogenin-3 expression.
abstract: Neurogenin-3 (ngn-3) expression is critical for endocrine development in the developing pancreas. We found that when ngn-3 was inhibited in an E11.5 pancreas, using either morpholino antisense or siRNA, it led to a significant decrease in endocrine differentiation after seven days in culture. Endocrine differentiation was rescued when ngn-3 inhibition was withdrawn after three days of culture, suggesting that the embryonic pancreas retains progenitor cells with the ability to differentiate into endocrine cell types when ngn-3 expression recurs. To determine whether the rescue phenomenon observed after withdrawing ngn-3 antisense treatment was the result of the original endocrine-committed cells reinitiating endocrine differentiation, or was instead due to new recruitment of later progenitor cells, we blocked ngn-3 expression for only the last four days of a seven-day culture. Here, insulin-positive differentiation was slightly reduced, but there was a normal number of glucagon-positive cells. In addition, there was an increase in SOX9-positive cells in ngn-3 inhibited, as well as in ngn-3 rescued pancreata, with a significant proportion of these SOX9-positive cells co-localized with DBA, an early ductal marker. This increased number of cells with co-localization of SOX9 and DBA could indicate an increased number of endocrine progenitor cells.
authors.affiliation: Division of Pediatric General and Thoracic Surgery, Children's Hospital of University of Pittsburgh Medical Center, One Children's Hospital Drive, 4401 Penn Ave., Rangos Research Center, Pittsburgh, PA 15224, USA. krishna.prasadan@chp.edu
authors.forename: Krishna
authors.initials: K
authors.lastname: Prasadan
authors.forename: Sidhartha
authors.initials: S
authors.lastname: Tulachan
authors.forename: Ping
authors.initials: P
authors.lastname: Guo
authors.forename: Chiyo
authors.initials: C
authors.lastname: Shiota
authors.forename: Sohail
authors.initials: S
authors.lastname: Shah
authors.forename: George
authors.initials: G
authors.lastname: Gittes
mesh_terms: D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics*; D002454:Cell Differentiation / Q000235:genetics*; D002478:Cells, Cultured; D005934:Glucagon / Q000378:metabolism; D007328:Insulin / Q000378:metabolism; D051379:Mice; D008815:Mice, Inbred Strains; D009419:Nerve Tissue Proteins / Q000235:genetics*; D046790:Pancreas, Exocrine / Q000166:cytology / Q000196:embryology*; D037121:Plant Lectins / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D055758:SOX9 Transcription Factor / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000502:physiology
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D005934:Glucagon; D007328:Insulin; D009419:Nerve Tissue Proteins; D037121:Plant Lectins; D034741:RNA, Small Interfering; D055758:SOX9 Transcription Factor; C407737:Neurog3 protein, mouse; C527933:Sox9 protein, mouse; C036554:dolichos biflorus agglutinin
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