Article

Characterization of the expression, localization, and secretion of PANDER in α-cells

Department of Pathology and Laboratory Medicine, The Children's Hospital of Philadelphia Research Institute, Philadelphia, PA 19104-4318, USA.
Molecular and Cellular Endocrinology (Impact Factor: 4.24). 08/2010; 325(1-2):36-45. DOI: 10.1016/j.mce.2010.05.008
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ABSTRACT The novel islet-specific protein PANcreatic DERived Factor (PANDER; FAM3B) has been extensively characterized with respect to the beta-cell, and these studies suggest a potential function for PANDER in the regulation of glucose homeostasis. Little is known regarding PANDER in pancreatic -cells, which are critically involved in maintaining euglycemia. Here we present the first report elucidating the expression and regulation of PANDER within the alpha-cell. Pander mRNA and protein are detected in alpha-cells, with primary localization to a glucagon-negative granular cytosolic compartment. PANDER secretion from alpha-cells is nutritionally and hormonally regulated by l-arginine and insulin, demonstrating similarities and differences with glucagon. Signaling via the insulin receptor (IR) through the PI3K and Akt/PKB node is required for insulin-stimulated PANDER release. The separate localization of PANDER and glucagon is consistent with their differential regulation, and the effect of insulin suggests a paracrine/endocrine effect on PANDER release. This provides further insight into the potential glucose-regulatory role of PANDER.

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    • "Arginine and, to a greater extent, insulin have been shown to induce PANDER expression in a-cells, but not in b-cells (Carnegie et al., 2010). In addition, in-vivo studies performed on the PANDER transgenic mouse model have indicated that PANDER expression is increased in the liver during fed conditions (Robert- Cooperman et al., 2014), whereas Li et al. has indicated that high fat diet may induce hepatic PANDER expression (Li et al., 2011). "
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    ABSTRACT: PANcreatic-DERived factor (PANDER, FAM3B) has been shown to regulate glycemic levels via interactions with both pancreatic islets and the liver. Although PANDER is predominantly expressed from the endocrine pancreas, recent work has provided sufficient evidence that the liver may also be an additional tissue source of PANDER production. At physiological levels, PANDER is capable of disrupting insulin signaling and promoting increased hepatic glucose production. As shown in some animal models, strong expression of PANDER, induced by viral delivery within the liver, induces hepatic steatosis. However, no studies to date have explicitly characterized the transcriptional regulation of PANDER from the liver. Therefore, our investigation elucidated the nutrient and hormonal regulation of the hepatic PANDER promoter. Initial RNA-ligated rapid amplification of cDNA ends identified a novel transcription start site (TSS) approximately 26 bp upstream of the PANDER translational start codon not previously revealed in pancreatic β-cell lines. Western evaluation of various murine tissues demonstrated robust expression in the liver and brain. Promoter analysis identified strong tissue-specific activity of the PANDER promoter in both human and murine liver-derived cell lines. The minimal element responsible for maximal promoter activity within hepatic cell lines was located between -293 to -3 of the identified TSS. PANDER promoter activity was inhibited by both insulin and palmitate, whereas glucose strongly increased expression. The minimal element was responsible for maximal glucose-responsive and basal activity. Co-transfection reporter assays, chromatin-immunoprecipitation (ChIP) and site-directed mutagenesis revealed that the carbohydrate-responsive element binding protein (ChREBP) increased PANDER promoter activity and interacted with the PANDER promoter. E-box 3 was shown to be critical for basal and glucose responsive expression. In summary, in-vitro and in-vivo glucose is a potent stimulator of the PANDER promoter within the liver and this response may be facilitated by ChREBP. Copyright © 2015. Published by Elsevier Ireland Ltd.
    Molecular and Cellular Endocrinology 06/2015; DOI:10.1016/j.mce.2015.05.040 · 4.24 Impact Factor
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    • "Glucose has been shown to significantly enhance Pander promoter activity and secretion from β cells and pancreatic islets (Burkhardt et al., 2005; Wang et al., 2008; Yang et al., 2005). In addition, PANDER is co-secreted with insulin in response to glucose (Carnegie et al., 2010; Xu et al., 2005). The mechanism of action and full biological effect of PANDER have yet to be fully elucidated in vivo, and this is the result of a lack of appropriate permanent and genomically integrated rodent models. "
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    ABSTRACT: PANcreatic-DERived Factor (PANDER, FAM3B) is a uniquely structured protein strongly expressed within and secreted from the endocrine pancreas. PANDER has been hypothesized to regulate fasting and fed glucose homeostasis, hepatic lipogenesis and insulin signaling, and serve a potential role in the onset or progression of type 2 diabetes. Despite having a potential pleiotropic pivotal role in glycemic regulation and T2D, there has been limited generation of stable animal models for PANDER investigation, with none on well-established genetic murine backgrounds for T2D. Our aim was to generate an enhanced murine model to further elucidate the biological function of PANDER. Therefore, a pure bred PANDER C57BL/6 knockout model (PANKO-C57) was created and phenotypically characterized with respect to glycemic regulation and hepatic insulin signaling. The PANKO-C57 exhibited an enhanced metabolic phenotype particularly with regard to enhanced glucose tolerance. Male PANKO-C57 mice displayed decreased fasting plasma insulin and c-peptide levels, whereas leptin levels were increased as compared to matched C57BL/6J WT mice. Despite similar peripheral insulin sensitivity between both groups, hepatic insulin signaling was significantly increased during fasting conditions as demonstrated by increased phosphorylation of hepatic Akt and AMPK along with mature SREBP-1 expression. Insulin stimulation of PANKO-C57 mice resulted in increased hepatic triglyceride and glycogen content as compared to C57BL/6 WT. In summary, the PANKO-C57 mouse represents a suitable model for the investigation of PANDER in multiple metabolic states and provides an additional tool to elucidate the biological function and potential role in T2D.
    Disease Models and Mechanisms 09/2014; 7(11). DOI:10.1242/dmm.016402 · 5.54 Impact Factor
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    • "With regard to biological function, in vitro studies revealed the increased production of PANDER mRNA and protein under glucose stimulation of the pancreatic b-cell, but not the a-cell (Burkhardt et al. 2005, Yang et al. 2005, Wang et al. 2008). In contrast, insulin stimulates PANDER secretion from the pancreatic a-TC1-6 cell line and appears to be located in an intracellular compartment distinct from glucagon (Carnegie et al. 2010). Palmitic acid has also been reported to induce PANDER mRNA and protein expression in a dose and time dependent manner in a pancreatic b-cell line (Chen et al. 2011, Xiang et al. 2012). "
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    ABSTRACT: PANcreatic-DERived factor (PANDER, FAM3B) is a novel protein that is highly expressed within the endocrine pancreas and to a lesser degree in other tissues. Under glucose stimulation, PANDER is co-secreted with insulin from the β-cell. Despite prior creation and characterization of acute hepatic PANDER animal models, the physiologic function remains to be elucidated from pancreas-secreted PANDER. To determine this, in this study, a transgenic mouse exclusively overexpressing PANDER from the endocrine pancreas was generated. PANDER was selectively expressed by the pancreatic-duodenal homeobox-1 (PDX1) promoter. The PANDER transgenic (PANTG) mice were metabolically and proteomically characterized to evaluate effects on glucose homeostasis, insulin sensitivity, and lipid metabolism. Fasting glucose, insulin and C-peptide levels were elevated in the PANTG compared with matched WT mice. Younger PANTG mice also displayed glucose intolerance in the absence of peripheral insulin sensitivity. Hyperinsulinemic-euglycemic clamp studies revealed that hepatic glucose production and insulin resistance were significantly increased in the PANTG with no difference in either glucose infusion rate or rate of disappearance. Fasting glucagon, corticosterones, resistin and leptin levels were also similar between PANTG and WT. Stable isotope labeling of amino acids in cell culture revealed increased gluconeogenic and lipogenic proteomic profiles within the liver of the PANTG with phosphoenol-pyruvate carboxykinase demonstrating a 3.5-fold increase in expression. This was matched with increased hepatic triglyceride content and decreased p-AMPK and p-acetyl coenzyme A carboxylase-1 signaling in the PANTG. Overall, our findings support a role of pancreatic β-cell-secreted PANDER in the regulation of hepatic insulin and lipogenenic signaling with subsequent impact on overall glycemia.
    Journal of Endocrinology 01/2014; 220(3):219-31. DOI:10.1530/JOE-13-0338 · 3.59 Impact Factor
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