A primer on modelling pancreatic islets: from models of coupled β-cells to multicellular islet models

Félix‐Martínez, Gerardo J.; Godínez‐Fernández, J. Rafael

doi:10.1080/19382014.2023.2231609

Cited by 3 publications

(1 citation statement)

References 127 publications

(158 reference statements)

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“…Because coordinated intercellular activity is not only crucial for tightly regulated insulin secretion but is also known to be altered in diabetes, researchers are investing considerable effort in describing and studying how collective rhythmicity is established in beta cell populations and how the underlying mechanisms change in disease. In recent years, the emergence of network analyses has provided a promising tool for evaluating data obtained through advanced multicellular imaging, with the goal to objectively characterize collective activity in islets [16,42,[54][55][56][57][58]. In this approach, individual cells serve as nodes, and their positions correspond to their physical locations within the tissue.…”

Section: Introductionmentioning

confidence: 99%

Network representation of multicellular activity in pancreatic islets: Technical considerations for functional connectivity analysis

Šterk,

Zhang,

Pohorec

et al. 2024

PLoS Comput Biol

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Within the islets of Langerhans, beta cells orchestrate synchronized insulin secretion, a pivotal aspect of metabolic homeostasis. Despite the inherent heterogeneity and multimodal activity of individual cells, intercellular coupling acts as a homogenizing force, enabling coordinated responses through the propagation of intercellular waves. Disruptions in this coordination are implicated in irregular insulin secretion, a hallmark of diabetes. Recently, innovative approaches, such as integrating multicellular calcium imaging with network analysis, have emerged for a quantitative assessment of the cellular activity in islets. However, different groups use distinct experimental preparations, microscopic techniques, apply different methods to process the measured signals and use various methods to derive functional connectivity patterns. This makes comparisons between findings and their integration into a bigger picture difficult and has led to disputes in functional connectivity interpretations. To address these issues, we present here a systematic analysis of how different approaches influence the network representation of islet activity. Our findings show that the choice of methods used to construct networks is not crucial, although care is needed when combining data from different islets. Conversely, the conclusions drawn from network analysis can be heavily affected by the pre-processing of the time series, the type of the oscillatory component in the signals, and by the experimental preparation. Our tutorial-like investigation aims to resolve interpretational issues, reconcile conflicting views, advance functional implications, and encourage researchers to adopt connectivity analysis. As we conclude, we outline challenges for future research, emphasizing the broader applicability of our conclusions to other tissues exhibiting complex multicellular dynamics.

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Section: Introductionmentioning

confidence: 99%

Network representation of multicellular activity in pancreatic islets: Technical considerations for functional connectivity analysis

Šterk,

Zhang,

Pohorec

et al. 2024

PLoS Comput Biol

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A practical and robust method to evaluate metabolic fluxes in primary pancreatic islets

Rocha,

Manucci,

Bruni-Cardoso

et al. 2024

Molecular Metabolism

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A practical and robust method to evaluate metabolic fluxes in pancreatic islets

Rocha

Manucci

Bruni‐Cardoso

et al. 2023

Preprint

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Aims/hypothesis: Efficient mitochondrial oxidative phosphorylation is essential for pancreatic beta cell responses to nutrient levels. Consequently, the evaluation of mitochondrial oxygen consumption and ATP production is important to investigate essential aspects of pancreatic islet pathophysiology. Currently, most studies use cell lines instead of primary islets due to difficulties in measuring primary islet respiration, which requires specific equipment and consumables that are expensive, complicated to use, and poorly reproducible. The aim of this study is to establish a robust and practical method to assess primary islet metabolic fluxes using Extracellular Flux Technology and standard commercial consumables. Methods: Pancreatic islets were isolated from 8 to 12-week-old mice and rats, and submitted to a dispersion protocol using trypsin. Dispersed islets were adhered overnight to pre-coated standard Seahorse microplates, and oxygen consumption rates were evaluated using a Seahorse Extracellular Flux Analyzer. We also validated the functionality of dispersed islets by analyzing glucose-stimulated insulin secretion (GSIS) and calcium (Ca2+) influx in response to different modulators by fluorescence microscopy. Results: We provide a detailed protocol with all steps necessary to optimize islet isolation and dispersion, in order to achieve a high yield of functional islets and perform metabolic flux analysis. With this method, which requires only a few islets per replicate, both rat and mouse islets present robust basal respiration and proper response to mitochondrial modulators (oligomycin, CCCP, antimycin and rotenone) and glucose addition. Both oligomycin and CCCP concentrations were titrated. Our method was also validated by other functional assays, which show these cells present conserved Ca2+ influx and insulin secretion in response to glucose. Conclusions/interpretation: We established a practical and robust method to assess ex vivo islet metabolic fluxes and oxidative phosphorylation. Our findings cover an important gap in primary islet physiology studies, providing a valuable tool we hope is useful to uncover basic beta cell metabolic mechanisms, as well as for translational investigations, such as pharmacological candidate discovery and islet transplantation protocols.

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A primer on modelling pancreatic islets: from models of coupled β-cells to multicellular islet models

Cited by 3 publications

References 127 publications

Network representation of multicellular activity in pancreatic islets: Technical considerations for functional connectivity analysis

Network representation of multicellular activity in pancreatic islets: Technical considerations for functional connectivity analysis

A practical and robust method to evaluate metabolic fluxes in primary pancreatic islets

A practical and robust method to evaluate metabolic fluxes in pancreatic islets

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