α-Cell Dysfunctions and Molecular Alterations in Male Insulinopenic Diabetic Mice Are Not Completely Corrected by Insulin

Dusaulcy, Rodolphe; Handgraaf, Sandra; Heddad-Masson, Mounia; Visentin, Florian; Vesin, Christian; Reimann, Frank; Gribble, Fiona M.; Philippe, Jacques; Gosmain, Yvan

doi:10.1210/en.2015-1725

Cited by 21 publications

(18 citation statements)

References 45 publications

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“…Moreover, cultured islet cells responded properly to the physiologically relevant secretagogues (high glucose for insulin, and arginine for glucagon), by inducing the respective hormone secretion about twofolds (Figure 4F). In line with these data, our recent work demonstrated that primary α-cells isolated from ProGcg-Venus mice responded to high glucose by a reduction in glucagon release (33), thus giving the opportunity to perform hormone secretion studies by these cells in vitro . This is particularly important for α-cells, since in mixed islet cell populations the glucagon secretion is altered by the amount of insulin secreted by adjacent β-cells, which represent the cell majority (34, 35).…”

Section: Discussionmentioning

confidence: 52%

High-Resolution Recording of the Circadian Oscillator in Primary Mouse α- and β-Cell Culture

2017

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Circadian clocks have been developed in evolution as an anticipatory mechanism allowing for adaptation to the constantly changing light environment due to rotation of the Earth. This mechanism is functional in all light-sensitive organisms. There is a considerable body of evidence on the tight connection between the circadian clock and most aspects of physiology and metabolism. Clocks, operative in the pancreatic islets, have caught particular attention in the last years due to recent reports on their critical roles in regulation of insulin secretion and etiology of type 2 diabetes. While β-cell clocks have been extensively studied during the last years, α-cell clocks and their role in islet function and orchestration of glucose metabolism stayed unexplored, largely due to the difficulty to isolate α-cells, which represents a considerable technical challenge. Here, we provide a detailed description of an experimental approach for the isolation of separate mouse α- and β-cell population, culture of isolated primary α- and β-cells, and their subsequent long-term high-resolution circadian bioluminescence recording. For this purpose, a triple reporter ProGlucagon-Venus/RIP-Cherry/Per2:Luciferase mouse line was established, carrying specific fluorescent reporters for α- and β-cells, and luciferase reporter for monitoring the molecular clockwork. Flow cytometry fluorescence-activated cell sorting allowed separating pure α- and β-cell populations from isolated islets. Experimental conditions, developed by us for the culture of functional primary mouse α- and β-cells for at least 10 days, will be highlighted. Importantly, temporal analysis of freshly isolated α- and β-cells around-the-clock revealed preserved rhythmicity of core clock genes expression. Finally, we describe the setting to assess circadian rhythm in cultured α- and β-cells synchronized in vitro. The here-described methodology allows to analyze the functional properties of primary α- and β-cells under physiological or pathophysiological conditions and to assess the islet cellular clock properties.

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Section: Discussionmentioning

confidence: 52%

High-Resolution Recording of the Circadian Oscillator in Primary Mouse α- and β-Cell Culture

2017

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“…Moreover, such system allows to dissect the temporal profiles of islet hormone secretion, along with the molecular characterization of the islet cellular oscillator . Of note, the recent development of highly specific fluorescent islet cell reporters has opened new horizons for studying separate α‐ and β‐cell functions in mice, and in humans …”

Section: Discussionmentioning

confidence: 76%

“…69 Moreover, such system allows to dissect the temporal profiles of islet hormone secretion, along with the molecular characterization of the islet cellular oscillator. 116 Of note, the recent development of highly specific fluorescent islet cell reporters has opened new horizons for studying separate αand β-cell functions in mice, 90,91,124 and in humans. 75,125 Additional challenge in translating clock studies from mouse/rat to human models is related to the peripheral clock differences of nocturnal vs. diurnal species.…”

Section: Cell-autonomous Circadian Oscillations In the Endocrine Pancmentioning

confidence: 99%

Time zones of pancreatic islet metabolism

Petrenko

Philippe

Dibner

2018

Diabetes Obesity Metabolism

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Most living beings possess an intrinsic system of circadian oscillators, allowing anticipation of the Earth's rotation around its own axis. The mammalian circadian timing system orchestrates nearly all aspects of physiology and behaviour. Together with systemic signals originating from the central clock that resides in the hypothalamic suprachiasmatic nucleus, peripheral oscillators orchestrate tissue-specific fluctuations in gene transcription and translation, and posttranslational modifications, driving overt rhythms in physiology and behaviour. There is accumulating evidence of a reciprocal connection between the circadian oscillator and most aspects of physiology and metabolism, in particular as the circadian system plays a critical role in orchestrating body glucose homeostasis. Recent reports imply that circadian clocks operative in the endocrine pancreas regulate insulin secretion, and that islet clock perturbation in rodents leads to the development of overt type 2 diabetes. While whole islet clocks have been extensively studied during the last years, the heterogeneity of islet cell oscillators and the interplay between α- and β-cellular clocks for orchestrating glucagon and insulin secretion have only recently gained attention. Here, we review recent findings on the molecular makeup of the circadian clocks operative in pancreatic islet cells in rodents and in humans, and focus on the physiologically relevant synchronizers that are resetting these time-keepers. Moreover, the implication of islet clock functional outputs in the temporal coordination of metabolism in health and disease will be highlighted.

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“…Murine primary cell preparation and cellular cultures. At the end of the protocol, primary α-Venus + and L cells and primary β-Cherry + cells from sham, ovx, ovx+v, and ovx+E 2 mice were separated from non-Venus and non-Cherry cells by FACS using Bio-Rad S3 and Moflo Astrios, after standard isolation procedures on the pancreas and small intestine, as described (50)(51)(52). FACS-purified α-, β-, and L cells were collected in acid/ethanol mixture (1.5% HCl/75% ethanol) for glucagon, insulin, and GLP-1 cellular content measurements or in RNAse-free lysis buffer (Qiagen RLT+ Buffer) for gene expression analyses.…”

Section: Methodsmentioning

confidence: 99%

17-β Estradiol regulates proglucagon-derived peptide secretion in mouse and human α- and L cells

Handgraaf¹,

Dusaulcy²,

Visentin³

et al. 2018

JCI Insight

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Clinical and experimental data indicate a beneficial effect of estrogens on energy and glucose homeostasis associated with improved insulin sensitivity and positive effects on insulin secretion. The aim of the study was to investigate the impact of estrogens on proglucagon-producing cells, pancreatic α cells, and enteroendocrine L cells. The consequences of sexual hormone deprivation were evaluated in ovariectomized mice (ovx). Ovx mice exhibited impaired glucose tolerance during oral glucose tolerance tests (OGTT), which was associated with decreased GLP-1 intestinal and pancreatic secretion and content, an effect that was reversed by estradiol (E2) treatment. Indeed, E2 increased oral glucose-induced GLP-1 secretion in vivo and GLP-1 secretion from primary culture of mouse and human α cells through the activation of all 3 estrogen receptors (ERs), whereas E2-induced GLP-1 secretion from mouse and human intestinal explants occurred only by ERβ activation. Underlying the implication of ERβ, its selective agonist WAY20070 was able to restore glucose tolerance in ovx mice at least partly through plasma GLP-1 increase. We conclude that E2 directly controls both α- and L cells to increase GLP-1 secretion, in addition to its effects on insulin and glucagon secretion, highlighting the potential beneficial role of the estrogenic pathway and, more particularly, of ERβ agonists to prevent type 2 diabetes.

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α-Cell Dysfunctions and Molecular Alterations in Male Insulinopenic Diabetic Mice Are Not Completely Corrected by Insulin

Cited by 21 publications

References 45 publications

High-Resolution Recording of the Circadian Oscillator in Primary Mouse α- and β-Cell Culture

High-Resolution Recording of the Circadian Oscillator in Primary Mouse α- and β-Cell Culture

Time zones of pancreatic islet metabolism

17-β Estradiol regulates proglucagon-derived peptide secretion in mouse and human α- and L cells

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