Dopamine Receptor Signaling in MIN6 β -Cells Revealed by Fluorescence Fluctuation Spectroscopy

Caldwell, Brittany; Ustione, Alessandro; Piston, David W.

doi:10.1016/j.bpj.2016.06.026

Cited by 6 publications

(7 citation statements)

References 58 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These cells are being widely used for preliminary investigations on beta‐cell biology and drug targets. For example, recent investigations include exploration of dopamine receptor signalling, lipidomic profiling, antidiabetic actions of dogfish glucagon analogues, high‐throughput screening for inhibitors of beta‐cell apoptosis, Ca 2+ channel clustering with insulin granules in diabetes, mitochondrial dynamics and bioenergetics as well as involvement of neuronal K + Cl − cotransporter in insulin secretion . These investigations lay a foundation for further studies on understanding of beta‐cell biology in health and disease.…”

Section: Development and Experimental Use Of Rodent Beta‐cell Linesmentioning

confidence: 99%

Cellular models for beta‐cell function and diabetes gene therapy

2018

View full text Add to dashboard Cite

Diabetes is characterized by the destruction and/or relative dysfunction of insulin-secreting beta-cells in the pancreatic islets of Langerhans. Consequently, considerable effort has been made to understand the physiological processes governing insulin production and secretion in these cells and to elucidate the mechanisms involved in their deterioration in the pathogenesis of diabetes. To date, considerable research has exploited clonal beta-cell lines derived from rodent insulinomas. Such cell lines have proven to be a great asset in diabetes research, in vitro drug testing, and studies of beta-cell physiology and provide a sustainable, and in many cases, more practical alternative to the use of animals or primary tissue. However, selection of the most appropriate rodent beta cell line is often challenging and no single cell line entirely recapitulates the properties of human beta-cells. The generation of stable human beta-cell lines would provide a much more suitable model for studies of human beta-cell physiology and pathology and could potentially be used as a readily available source of implantable insulin-releasing tissue for cell-based therapies of diabetes. In this review, we discuss the history, development, functional characteristics and use of available clonal rodent beta-cell lines, as well as reflecting on recent advances in the generation of human-derived beta-cell lines, their use in research studies and their potential for cell therapy of diabetes.

show abstract

Section: Development and Experimental Use Of Rodent Beta‐cell Linesmentioning

confidence: 99%

Cellular models for beta‐cell function and diabetes gene therapy

2018

View full text Add to dashboard Cite

show abstract

“…Clinical studies have reported that patients with Parkinson's disease who were treated with the long-term dopamine precursor L-dopa had a significant reduction in glucose tolerance and even developed type 2 diabetes (9). In recent years, dopamine has been proposed to be important in regulating glucose-stimulated insulin secretion (10,11). However, the characteristic effect of dopamine on insulin secretion and the underlying electrophysiological mechanism have not been extensively studied.…”

Section: Introductionmentioning

confidence: 99%

D2-Like Receptors Mediate Dopamine-Inhibited Insulin Secretion via Ion Channels in Rat Pancreatic β-Cells

et al. 2020

View full text Add to dashboard Cite

Dopamine (DA) has a vital role in the central nervous system and also modulates lipid and glucose metabolism. The present study aimed to investigate the effect of dopamine on insulin secretion and the underlying mechanisms in rat pancreatic β-cells. Data from the radioimmunoassay indicated that dopamine inhibited insulin secretion in a glucoseand dose-dependent manner. This inhibitory effect of dopamine was mediated mainly by D2-like receptors, but not D1-like receptors. Whole-cell patch-clamp recordings showed that dopamine decreased voltage-dependent Ca 2+ channel currents, which could be reversed by inhibition of the D2-like receptor. Dopamine increased voltage-dependent potassium (K V) channel currents and shortened action potential duration, which was antagonized by inhibition of D2-like receptors. Further experiments showed that D2-like receptor activation by quinpirole increased K V channel currents. In addition, using calcium imaging techniques, we found that dopamine reduced intracellular Ca 2+ concentration, which was also reversed by D2-like receptor antagonists. Similarly, quinpirole was found to decrease intracellular Ca 2+ levels. Taken together, these findings demonstrate that dopamine inhibits insulin secretion mainly by acting on D2-like receptors, inhibiting Ca 2+ channels, and activating Kv channels. This process results in shortened action potential duration and decreased intracellular Ca 2+ levels in β-cells. This work offers new insights into a glucose-dependent mechanism whereby dopamine regulates insulin secretion.

show abstract

“…Construction of pmApple-GNG2 was described previously (17). pEGFP-DRD2 was obtained from Jean-Michel Arrang via Addgene (#24099) (18).…”

Section: Plasmidsmentioning

confidence: 99%

Two-Color Spatial Cumulant Analysis Detects Heteromeric Interactions between Membrane Proteins

Foust

Godin

Ustione³

et al. 2019

Preprint

View full text Add to dashboard Cite

Fluorescence fluctuation spectroscopy (FFS) can be used to measure the aggregation of fluorescently labeled molecules and is typically performed using time series data. Spatial intensity distribution analysis (SpIDA) and fluorescence moment image analysis (FMIA) are established tools for measuring molecular brightnesses from single-color images collected with laser scanning microscopes. We have extended these tools for analysis of two-color images to resolve heteromeric interactions between molecules labeled with spectrally distinct chromophores. We call these new methods two-color SpIDA (2c-SpIDA) and two-color spatial cumulant analysis (2c-SpCA). To implement these techniques on a hyperspectral imaging system, we developed a spectral shift filtering (SSF) technique to remove artifacts due to intrinsic crosstalk between detector bins. We determined that 2c-SpCA provides better resolution from samples containing multiple fluorescent species, hence this technique was carried forward to study images of living cells. We used fluorescent heterodimers labeled with EGFP and mApple to quantify the effects of resonance energy transfer and incomplete maturation of mApple on brightness measurements. We show that 2c-SpCA can detect the interaction between two components of trimeric G-protein complexes. Thus 2c-SpCA presents a robust and computationally expedient means of measuring heteromeric interactions in cellular environments. Statement of SignificanceFluorescence fluctuation spectroscopy (FFS) techniques determine biophysical parameters from samples containing fluorescently labeled biomolecules by considering the statistical nature of fluorescent signals measured with photodetectors. The present study introduces two-color spatial cumulant analysis (2c-SpCA) to the canon of FFS techniques. 2c-SpCA analyzes pixel-value data of two-color images collected with laser scanning fluorescence microscopes. We show that 2c-SpCA can determine several biophysical parameters in living cells including Forster resonance energy transfer efficiency, the dark state fraction of fluorescent proteins, and heteromerization between distinctly labeled proteins. In comparison to existing techniques, 2c-SpCA requires very few image frames for analysis, minimal computations, and can be applied to images of fixed tissue samples.

show abstract

Dopamine Receptor Signaling in MIN6 β -Cells Revealed by Fluorescence Fluctuation Spectroscopy

Cited by 6 publications

References 58 publications

Cellular models for beta‐cell function and diabetes gene therapy

Cellular models for beta‐cell function and diabetes gene therapy

D2-Like Receptors Mediate Dopamine-Inhibited Insulin Secretion via Ion Channels in Rat Pancreatic β-Cells

Two-Color Spatial Cumulant Analysis Detects Heteromeric Interactions between Membrane Proteins

Contact Info

Product

Resources

About