Transcriptomic profiling of pancreatic alpha, beta and delta cell populations identifies delta cells as a principal target for ghrelin in mouse islets

Adriaenssens, Alice E.; Svendsen, Berit; Lam, Brian; Yeo, Giles S.H.; Holst, Jens J.; Reimann, Frank; Gribble, Fiona M.

doi:10.1007/s00125-016-4033-1

Cited by 169 publications

(247 citation statements)

References 48 publications

Supporting

Mentioning

217

Contrasting

Order By: Relevance

“…In an agreement with the previous studies, we demonstrate that a reporter-based separation of endocrine cells from pancreatic islets allows to obtain α- and β-cell populations bearing high expression levels of glucagon , and insulin and MafA transcripts, respectively (Figures 4A,B) (30, 31). Importantly, recently published by us circadian RNA sequencing analysis of thus separated α- and β-cell populations provides further extensive characterization of their differential transcriptional patterns (32).…”

Section: Discussionsupporting

confidence: 92%

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

2017

View full text Add to dashboard Cite

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.

show abstract

Section: Discussionsupporting

confidence: 92%

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

2017

View full text Add to dashboard Cite

show abstract

“…Insulin receptor exhibited similar expression levels and temporal patterns in both cell types according to our data ( Fig. 5; Supplemental Data Set 1) and previously published data sets (Benner et al 2014;Adriaenssens et al 2016;DiGruccio et al 2016). In response to a high dose of insulin, α-cellular and β-cellular clocks exhibit a similar circadian phase ( Fig.…”

Section: Discussionsupporting

confidence: 51%

Pancreatic α- and β-cellular clocks have distinct molecular properties and impact on islet hormone secretion and gene expression

et al. 2017

View full text Add to dashboard Cite

A critical role of circadian oscillators in orchestrating insulin secretion and islet gene transcription has been demonstrated recently. However, these studies focused on whole islets and did not explore the interplay between α-cell and β-cell clocks. We performed a parallel analysis of the molecular properties of α-cell and β-cell oscillators using a mouse model expressing three reporter genes: one labeling α cells, one specific for β cells, and a third monitoring circadian gene expression. Thus, phase entrainment properties, gene expression, and functional outputs of the α-cell and β-cell clockworks could be assessed in vivo and in vitro at the population and single-cell level. These experiments showed that α-cellular and β-cellular clocks are oscillating with distinct phases in vivo and in vitro. Diurnal transcriptome analysis in separated α and β cells revealed that a high number of genes with key roles in islet physiology, including regulators of glucose sensing and hormone secretion, are differentially expressed in these cell types. Moreover, temporal insulin and glucagon secretion exhibited distinct oscillatory profiles both in vivo and in vitro. Altogether, our data indicate that differential entrainment characteristics of circadian α-cell and β-cell clocks are an important feature in the temporal coordination of endocrine function and gene expression.

show abstract

“…First, Pax6 deletion causes a massive loss of β cell identity (potentially in addition to increased β cell death), leading to hypoinsulinemia and hyperglycemia. Second, ghrelin secretion from mutant β cells may further inhibit insulin secretion from remaining functional β cells via mechanisms proposed elsewhere (65)(66)(67). We note, however, that thus far, there is no evidence for expression of ghrelin in islets of human patients with diabetes.…”

Section: Pax6 Directly Regulates Transcription From the Insulin Gene mentioning

confidence: 73%

PAX6 maintains β cell identity by repressing genes of alternative islet cell types

Swisa¹,

Avrahami²,

Eden³

et al. 2016

Journal of Clinical Investigation

139

142

View full text Add to dashboard Cite

Transcriptomic profiling of pancreatic alpha, beta and delta cell populations identifies delta cells as a principal target for ghrelin in mouse islets

Cited by 169 publications

References 48 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

Pancreatic α- and β-cellular clocks have distinct molecular properties and impact on islet hormone secretion and gene expression

PAX6 maintains β cell identity by repressing genes of alternative islet cell types

Contact Info

Product

Resources

About