Homeobox Gene Product Nkx 6.1 Immunoreactivity in Nuclei of Endocrine Cells of Rat and Mouse Stomach

Øster, Anne; Jensen, Jan; Edlund, Helena; Larsson, Lars

doi:10.1177/002215549804600603

Cited by 53 publications

(41 citation statements)

References 15 publications

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“…This gene appears initially in almost all of the epithelial cells of the pancreatic buds, until around embryonic day 13 (e13), where its expression becomes restricted to in beta cells [19]. Even though this transcription factor may appear in other tissues, such as the stomach [8], we believe that Nkx6.1 would be a very good candidate gene for our construct, provided that cells are selected after directed differentiation. We also included a hygromycin resistance gene in the plasmid, which is expressed continuously in the cell by a pGK promoter.…”

Section: Mes Cells Differentiate Towards Insulin-and Pdx-1-positive Cmentioning

confidence: 99%

“…For cell trapping, the Nkx6.1 gene promoter was used because this transcription factor is required for the expansion of beta cell precursors and is selectively expressed in beta cells in the adult pancreas [6,7]. It also appears in the enterochromaffin cells of the antropyloric mucosa of the stomach [8], but in this region there is no Pdx-1 or insulin expression, whereas in beta cells these three genes are expressed. To induce differentiation, in contrast with previous attempts [9], differentiation factors were added during the embryoid body formation.…”

Section: Introductionmentioning

confidence: 99%

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In vitro directed differentiation of mouse embryonic stem cells into insulin-producing cells

et al. 2004

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Aims/hypothesis. We recently demonstrated that insulin-producing cells derived from embryonic stem cells normalise hyperglycaemia in transplanted diabetic mice. The differentiation and selection procedure, however, was successful in less than 5% of the assays performed. Thus, to improve its effectiveness, new strategies have been developed, which increase the number of islet cells or islet progenitors. Methods. Mouse embryonic stem cells transfected with a plasmid containing the Nkx6.1 promoter gene followed by a neomycin-resistance gene, were cultured with factors known to participate in endocrine pancreatic development and factors that modulate signalling pathways involved in these processes. Neomycin was used to select the Nkx6.1-positive cells, which also express insulin. The transfected cells were differentiated using several exogenous agents, followed by selection of Nkx6.1-positive cells. The resulting cells were analysed for pancreatic gene and protein expression by immunocytochemistry, RT-PCR and radioimmunoassay. Also, proliferation assays were performed, as well as transplantation to streptozotocin-induced diabetic mice. Results. The protocols yielded cell cultures with approximately 20% of cells co-expressing insulin and Pdx-1. Cell trapping selection yielded an almost pure population of insulin-positive cells, which expressed the beta cell genes/proteins Pdx-1, Nkx6.1, insulin, glucokinase, GLUT-2 and Sur-1. Subsequent transplantation to streptozotocin-induced diabetic mice normalised their glycaemia during the time period of experimentation, proving the efficiency of the protocols. Conclusions/interpretation. These methods were both highly efficient and very reproducible, resulting in a new strategy to obtain insulin-containing cells from stem cells with a near 100% success rate, while actively promoting the maturation of the exocytotic machinery.Keywords Diabetes · Differentiation · Embryonic stem cells · Exogenous agents · Insulin-producing cells · Islet progenitors Abbreviations: Anti-Shh, antibody against sonic hedgehog · D3, undifferentiated D3 stem cell line · EB, embryoid bodies · ES, embryonic stem · FBS, fetal bovine serum · LIF, leukaemia inhibitory factor · mES, mouse embryonic stem · Ngn3, neurogenin 3 · P, gelatine-coated plates · Pdx-1, pancreatic duodenum homeobox 1

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Section: Mes Cells Differentiate Towards Insulin-and Pdx-1-positive Cmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

In vitro directed differentiation of mouse embryonic stem cells into insulin-producing cells

et al. 2004

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“…173 Nkx6.1 is observed at e9.5 in both pancreatic buds until e13 and is then confined to the developing b-cells. 174,175 Although Nkx6.1 deficient mice do not exhibit b-cells, the other cell subtypes develop normally (Table 1). 175 In the b-cells, Nkx6.1 inhibits glucagon promoter activity.…”

Section: Islet 1 (Isl1)mentioning

confidence: 99%

Transcription factor regulation of pancreatic organogenesis, differentiation and maturation

Dassaye

Naidoo

Cerf

2015

Islets

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Lineage tracing studies have revealed that transcription factors play a cardinal role in pancreatic development, differentiation and function. Three transitions define pancreatic organogenesis, differentiation and maturation. In the primary transition, when pancreatic organogenesis is initiated, there is active proliferation of pancreatic progenitor cells. During the secondary transition, defined by differentiation, there is growth, branching, differentiation and pancreatic cell lineage allocation. The tertiary transition is characterized by differentiated pancreatic cells that undergo further remodeling, including apoptosis, replication and neogenesis thereby establishing a mature organ. Transcription factors function at multiple levels and may regulate one another and auto-regulate. The interaction between extrinsic signals from non-pancreatic tissues and intrinsic transcription factors form a complex gene regulatory network ultimately culminating in the different cell lineages and tissue types in the developing pancreas. Mutations in these transcription factors clinically manifest as subtypes of diabetes mellitus. Current treatment for diabetes is not curative and thus, developmental biologists and stem cell researchers are utilizing knowledge of normal pancreatic development to explore novel therapeutic alternatives. This review summarizes current knowledge of transcription factors involved in pancreatic development and b-cell differentiation in rodents.

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“…Nkx6.1 is not expressed in Pdx1-deficient mice, indicating that Pdx1 acts upstream of Nkx6.1 in enteroendocrine differentiation. 164 Some enteroendocrine hormone-producing intestinal populations are reduced in Nkx2.2-deficient mice, including serotonin-, CCK-, GIP-, and gastrinproducing cells; however, the number of ghrelin-expressing cells is greater, and the numbers of Paneth cells, goblet cells, and enterocytes are unaltered. It seems that Nkx2.2 functions upstream of Pax6 in regulating the fate of intestinal cells, as Pax6 mRNA and protein levels are reduced in Nkx2.2-deficient cells.…”

Section: Cell Differentiation and Intestinal Adaptationmentioning

confidence: 99%

Isolated ileal interposition in enteroendocrine L cells differentiation

et al. 2011

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INTRODUCTION:Due to the progressive nature of type 2 diabetes, its complexity and drug treatment perpetuity, there is currently a search for surgical procedures that can promote euglycemia also in non-obese patients. Diabetic patients glycemic control can be achieved by increasing the blood concentration of GLP-1, a hormone produced by L cells that are more densely concentrated in the terminal ileum. Early and extended improvement of diabetes in patients submitted to bariatric surgeries awakened the necessity of investigating the isolated ileal interposition as surgical alternative for the treatment of diabetes. The interposition of this ileal segment to a more anterior region (proximal jejunum) can promote a greater stimulation of the L cells by poorly digested food, increasing the production of GLP-1 and reflecting on glycemic control. However, in order to consolidate the ileal interposition as a surgical treatment of diabetes it is necessary that the interposed ileum keep the same differentiation rate into L cells for a long period to justify the intervention. AIMS:To investigate the isolated ileal interposition influence on the differentiation of intestinal precursor cells into enteroendocrine L cells over time. METHODS:Twelve 12-week-old male Wistar rats (Rattus norvegicus albinus) of the WAB strain (heterogeneous) will be used. All animals will receive a high-calorie, high-fat diet for 16 weeks or more until they develop glucose dysmetabolism confirmed by glycemic test. They will be divided into two groups of 10 animals each: the isolated ileal interposition group (GI) and the control group (GC), comprising animals that will not be submitted to any surgical intervention.Blood samples will be collected under anesthesia at the weeks 12, 26, 36 and 44 for the determination of serum levels of glucose, insulin, GLP-1, glucagon, C-peptide and glycosilated hemoglobin. The insulin tolerance test will be performed and insulin resistance will be calculated. For the comparative analysis of the ileal precursor cells differentiation into enteroendocrine cells among the two groups, the following intestinal fragments will be collected after euthanasia: interposed ileum and remaining ileum from GI, jejunum and ileum from GC. These fragments will be analyzed by imunofluorescence and also by Real Time PCR using PCR Arrays for target genes including the main ones related to stem cell, stem cell singnalling, diabetes, Wnt and Notch signaling pathways and other genes and pathways involved in the differentiation of intestinal precursor cells into enteroendocrine cells, especially GLP-1-producing L cells that play important role in euglycemia.

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Homeobox Gene Product Nkx 6.1 Immunoreactivity in Nuclei of Endocrine Cells of Rat and Mouse Stomach

Cited by 53 publications

References 15 publications

In vitro directed differentiation of mouse embryonic stem cells into insulin-producing cells

In vitro directed differentiation of mouse embryonic stem cells into insulin-producing cells

Transcription factor regulation of pancreatic organogenesis, differentiation and maturation

Isolated ileal interposition in enteroendocrine L cells differentiation

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