Conversion of amylase-secreting rat pancreatic AR42J cells to neuronlike cells by activin A.

Ohnishi, Hirohide; Ohgushi, Naohiro; Tanaka, Shigeyasu; Mogami, Hideo; Nobusawa, Romi; Mashima, Hirosato; Furukawa, Megumi; Mine, Tetsuya; Shimada, Osamu; Ishikawa, Hiroko

doi:10.1172/jci117922

Cited by 40 publications

(42 citation statements)

References 45 publications

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“…For example, two of the genes up-regulated during activin A-induced differentiation of AR42J cells to neuron-like cells include the intermediate neurofilament gene and the synaptophysin gene (15). Both genes are known targets of NRSF (11).…”

Section: Resultsmentioning

confidence: 99%

Regulation of Pax4 Paired Homeodomain Gene by Neuron-restrictive Silencer Factor

Kemp

Lin

Habener

2003

Journal of Biological Chemistry

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An elucidation of the key regulatory factors in pancreas development is critical for understanding the pathogenesis of diabetes mellitus. This study examined whether a specific regulatory mechanism that exists in neuronal development also plays a role in the pancreas. In non-neuronal cells, neuron-restrictive silencer factor (NSRF) actively represses gene transcription via a sequence-specific DNA motif known as the neuron-restrictive silencer element (NRSE). This DNA motif has been identified in many genes that are specific markers for cells of neuronal and neuroendocrine lineage. We identified several genes involved in pancreas development that also harbor NRSE-like motifs, including pdx-1, Beta2/NeuroD, and pax4. The paired homeodomain transcription factor Pax4 is implicated in the differentiation of the insulin-producing ␤-cell lineage because disruption of the pax4 gene results in a severe deficiency of ␤-cells and the manifestation of diabetes mellitus in mice. The NRSE-like motif identified in the upstream pax4 promoter is highly conserved throughout evolution, forms a DNA-protein complex with NRSF, and confers NRSF-dependent transcriptional repression in the context of a surrogate gene promoter. This cis-activating NRSE element also confers NRSF-dependent modulation in the context of the native pax4 gene promoter. Together with earlier reports, these new findings suggest an important functional role for NRSF in the expression of the pax4 gene and infer a role for NRSF in pancreatic islet development.Pancreas development appears to follow a precise pattern of gene expression events characterized by temporal and spatial specificity. As such, a hierarchical model of transcription factor gene expression has evolved based on phenotypic observations from specific knock-out and transgenic animal studies. An interpretation of these data is illustrated in Fig. 1 (see "Results") (1). Several transcription factors that are essential for ␤-cell differentiation are also involved in the neuronal development program. For example, the expression of Beta2⁄NeuroD is necessary for terminal differentiation of all endocrine cell types similar to its requirement for the differentiation of neurons in the brain (1). Similarities between islet and neuronal cells also exist at the level of physiology, such as in their electrical excitability and secretory vesicle functions. In addition to these similarities in differentiation and phenotypic profiles, the likelihood of the existence of additional molecular parallels is strong. Here we investigated whether the NRSE/NRSF 1 transcriptional repressor mechanism is involved in islet cell gene expression coincident with its role in neuron-specific gene regulation.The cis-regulatory NRSE is a 21-bp motif that confers transcriptional repression of genes in many non-neuronal cell types (2, 3). Inhibition of NRSF binding to the NRSE in differentiating neurons permits transcriptional expression of these genes and facilitates terminal differentiation of neuronal lineages through neuron-specific...

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Section: Resultsmentioning

confidence: 99%

Regulation of Pax4 Paired Homeodomain Gene by Neuron-restrictive Silencer Factor

Kemp

Lin

Habener

2003

Journal of Biological Chemistry

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“…Competitive Reverse Transcription-PCR of TGF-␤ 1 mRNA-Total RNA was obtained from PCS cells by using ISOGEN (Wako, Tokyo, Japan), followed with synthesis of double-stranded DNA as described previously (18). Competitive PCR of TGF-␤ 1 mRNA was performed using the competitive PCR kit for rat TGF-␤ 1 (Maxim Biotech Inc., San Francisco, CA) according to the manufacturer's instructions.…”

Section: Methodsmentioning

confidence: 99%

Distinct Roles of Smad2-, Smad3-, and ERK-dependent Pathways in Transforming Growth Factor-β1 Regulation of Pancreatic Stellate Cellular Functions

Ohnishi

Miyata

Yasuda

et al. 2004

Journal of Biological Chemistry

102

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Pancreatic stellate cells (PSCs) play a major role in promoting pancreatic fibrosis. Transforming growth factor-␤ 1 (TGF-␤ 1 ) regulates PSC activation and proliferation in an autocrine manner. The intracellular signaling pathways of the regulation were examined in this study. Immunoprecipitation and immunocytochemistry revealed that Smad2, Smad3, and Smad4 were functionally expressed in PSCs. Adenovirus-mediated expression of Smad2, Smad3, or dominant-negative Smad2/3 did not alter TGF-␤ 1

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“…In addition, activin-treated cells express mRNA for GLUT2, ATP-sensitive potassium channel, and pancreatic polypeptide (PP). Thus, activin A converts AR42J cells into endocrine cells (13,14). Furthermore, in the presence of betacellulin (14) or hepatocyte growth factor (HGF) (15), activin-treated AR42J cells further convert to insulin-producing cells.…”

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confidence: 99%

Changes in the expression of transcription factors in pancreatic AR42J cells during differentiation into insulin-producing cells.

2001

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Pancreatic AR42J cells possess both exocrine and neuroendocrine properties and convert to insulin-producing cells upon treatment with activin A and hepatocyte growth factor (HGF). We studied changes in the mRNA expression of various transcription factors during the course of differentiation. Among the transcription factors studied, expression levels of Pax4 and neurogenin3 changed significantly. These two factors were not detected in naive cells, whereas their mRNA levels were markedly increased after treatment with activin A and HGF. Thus, these two factors were induced by activin A. P ancreatic endocrine and exocrine cells are thought to arise from common precursor cells located in the epithelium of pancreatic anlage during development (1-3). Recent genetic studies indicate that pancreatic development depends on an integrated network of distinct transcription factors operating at various levels. Several homeodomain and basic helix-loop-helix transcription factors have been postulated to play important roles in regulating differentiation of pancreatic endocrine cells. A mouse homeobox protein, insulin promoter factor-1 (IPF-1/PDX-1), is required for development of the murine pancreas (4,5). Islet-1 (Isl-1), a LIM homeodomain-containing protein, is necessary for the development of the dorsal pancreas and is required for generation of islet cells (6). In addition, inactivation of Beta-2/ NeuroD causes a striking reduction in the number of insulinproducing cells, and mice lacking the functional Pax4 gene lack differentiated ␤-and ␦-cells and fail to develop mature islets (7,8). In contrast, the number of all differentiated endocrine cell types, especially the number of ␣-cells, is markedly reduced in mice lacking the Pax6 gene (9). These transcription factors are thought to be involved in a complex regulatory network and cascade to exert their functions (10,11). However, because differentiation of pancreatic ␤-cells involves many steps, it is not clear how these steps are regulated by transcription factors.Pancreatic AR42J cells are derived from a chemically induced pancreatic tumor and express both exocrine and neuroendocrine properties (12). Upon treatment with activin A, AR42J cells stop growing, and their morphology changes significantly by extending neurites (13). In addition, activin-treated cells express mRNA for GLUT2, ATP-sensitive potassium channel, and pancreatic polypeptide (PP). Thus, activin A converts AR42J cells into endocrine cells (13,14). Furthermore, in the presence of betacellulin (14) or hepatocyte growth factor (HGF) (15), activin-treated AR42J cells further convert to insulin-producing cells. Thus, AR42J cells provide a model system to study the molecular mechanism involved in ␤-cell differentiation. The present study was conducted to investigate how the expression of islet-associated transcription factors is changed during the differentiation of AR42J cells. We also assessed the role of such transcription factors in differentiation. RESEARCH DESIGN AND METHODS Materials.Recombinant human a...

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Conversion of amylase-secreting rat pancreatic AR42J cells to neuronlike cells by activin A.

Cited by 40 publications

References 45 publications

Regulation of Pax4 Paired Homeodomain Gene by Neuron-restrictive Silencer Factor

Regulation of Pax4 Paired Homeodomain Gene by Neuron-restrictive Silencer Factor

Distinct Roles of Smad2-, Smad3-, and ERK-dependent Pathways in Transforming Growth Factor-β1 Regulation of Pancreatic Stellate Cellular Functions

Changes in the expression of transcription factors in pancreatic AR42J cells during differentiation into insulin-producing cells.

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