Visualizing subcellular rearrangements in intact β cells using soft x-ray tomography

Subcellular maps of β cells reveal the coordinated reorganization of organelles during insulin secretion. Characterizing relationships between cell structures and functions requires mesoscale mapping of intact cells showing subcellular rearrangements following stimulation; however, current approaches are limited in this regard. Here, we report a unique application of soft x-ray tomography to generate three-dimensional reconstructions of whole pancreatic β cells at different time points following glucose-stimulated insulin secretion. Reconstructions following stimulation showed distinct insulin vesicle distribution patterns reflective of altered vesicle pool sizes as they travel through the secretory pathway. Our results show that glucose stimulation caused rapid changes in biochemical composition and/or density of insulin packing, increased mitochondrial volume, and closer proximity of insulin vesicles to mitochondria. Costimulation with exendin-4 (a glucagon-like peptide-1 receptor agonist) prolonged these effects and increased insulin packaging efficiency and vesicle maturation. This study provides unique perspectives on the coordinated structural reorganization and interactions of organelles that dictate cell responses.

[1]  R. Stevens,et al.  Visualizing insulin vesicle neighborhoods in β cells by cryo–electron tomography , 2020, Science Advances.

[2]  B. Giepmans,et al.  Large-scale electron microscopy database for human type 1 diabetes , 2020, Nature Communications.

[3]  A. Dlasková,et al.  Overexpression of native IF1 downregulates glucose-stimulated insulin secretion by pancreatic INS-1E cells , 2020, Scientific Reports.

[4]  M. Aronova,et al.  Determination of secretory granule maturation times in pancreatic islet beta-cells by serial block face scanning electron microscopy , 2019, bioRxiv.

[5]  S. Stanbury.  Visualizing , 2019, A New Companion to Chaucer.

[6]  T. Rapoport,et al.  Faculty Opinions recommendation of Visualizing intracellular organelle and cytoskeletal interactions at nanoscale resolution on millisecond timescales. , 2019, Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature.

[7]  M. Ailion,et al.  EIPR1 controls dense-core vesicle cargo retention and EARP complex localization in insulin-secreting cells , 2018, bioRxiv.

[8]  J. Lippincott-Schwartz,et al.  Visualizing Intracellular Organelle and Cytoskeletal Interactions at Nanoscale Resolution on Millisecond Timescales , 2018, Cell.

[9]  Gregory R. Johnson,et al.  Label-free prediction of three-dimensional fluorescence images from transmitted light microscopy , 2018, Nature Methods.

[10]  Frank Alber,et al.  Opportunities and Challenges in Building a Spatiotemporal Multi-scale Model of the Human Pancreatic β Cell , 2018, Cell.

[11]  G. McDermott,et al.  PSF correction in soft x-ray tomography , 2018, bioRxiv.

[12]  P. Drain,et al.  Glucose but not KCl diminishes submembrane granule turnover in mouse beta-cells. , 2017, Journal of molecular endocrinology.

[13]  Devin P. Sullivan,et al.  A subcellular map of the human proteome , 2017, Science.

[14]  D. Fu,et al.  Coupling of Insulin Secretion and Display of a Granule-resident Zinc Transporter ZnT8 on the Surface of Pancreatic Beta Cells* , 2017, The Journal of Biological Chemistry.

[15]  Gerry McDermott,et al.  Mesoscale imaging with cryo‐light and X‐rays: Larger than molecular machines, smaller than a cell , 2017, Biology of the cell.

[16]  Manolis Kellis,et al.  Soft X-Ray Tomography Reveals Gradual Chromatin Compaction and Reorganization during Neurogenesis In Vivo. , 2016, Cell reports.

[17]  C. Verchere,et al.  Glucagon-Like Peptide-1 Receptor Agonists: Beta-Cell Protection or Exhaustion? , 2016, Trends in Endocrinology & Metabolism.

[18]  J. Cox,et al.  Global, quantitative and dynamic mapping of protein subcellular localization , 2016, eLife.

[19]  A. Hyman,et al.  Visualizing the molecular sociology at the HeLa cell nuclear periphery , 2016, Science.

[20]  Y. Kalaidzidis,et al.  A Spatial Model of Insulin‐Granule Dynamics in Pancreatic β‐Cells , 2015, Traffic.

[21]  Y. Cho,et al.  Glucagon-Like Peptide-1 Increases Mitochondrial Biogenesis and Function in INS-1 Rat Insulinoma Cells , 2015, Endocrinology and metabolism.

[22]  A. Burnside,et al.  Activins A and B Regulate Fate-Determining Gene Expression in Islet Cell Lines and Islet Cells From Male Mice. , 2015, Endocrinology.

[23]  D. Goodsell,et al.  cellPACK: A Virtual Mesoscope to Model and Visualize Structural Systems Biology , 2014, Nature Methods.

[24]  Gerry McDermott,et al.  Biological soft X-ray tomography on beamline 2.1 at the Advanced Light Source. , 2014, Journal of synchrotron radiation.

[25]  Michael Krauss,et al.  Composition of isolated synaptic boutons reveals the amounts of vesicle trafficking proteins , 2014, Science.

[26]  M. Grabenbauer,et al.  Golgi apparatus analyzed by cryo-electron microscopy , 2013, Histochemistry and Cell Biology.

[27]  C. Burant,et al.  Metabolome Response to Glucose in the β-Cell Line INS-1 832/13* , 2013, The Journal of Biological Chemistry.

[28]  D. Turnbull,et al.  Mitochondrial morphology, topology, and membrane interactions in skeletal muscle: a quantitative three-dimensional electron microscopy study. , 2013, Journal of applied physiology.

[29]  Carolyn A. Larabell,et al.  Nuclear Aggregation of Olfactory Receptor Genes Governs Their Monogenic Expression , 2012, Cell.

[30]  Chao Yang,et al.  Automatic alignment and reconstruction of images for soft X-ray tomography. , 2012, Journal of structural biology.

[31]  Ben M. Webb,et al.  Putting the Pieces Together: Integrative Modeling Platform Software for Structure Determination of Macromolecular Assemblies , 2012, PLoS biology.

[32]  A. Müller,et al.  Novel standards in the measurement of rat insulin granules combining electron microscopy, high-content image analysis and in silico modelling , 2012, Diabetologia.

[33]  M. Rupnik,et al.  Pancreatic beta cell lines and their applications in diabetes mellitus research. , 2010, ALTEX.

[34]  Gerry McDermott,et al.  Soft X-ray tomography and cryogenic light microscopy: the cool combination in cellular imaging. , 2009, Trends in cell biology.

[35]  M. Granvik,et al.  Insulin crystallization depends on zinc transporter ZnT8 expression, but is not required for normal glucose homeostasis in mice , 2009, Proceedings of the National Academy of Sciences.

[36]  J. Pessin,et al.  Insulin granule biogenesis, trafficking and exocytosis. , 2009, Vitamins and hormones.

[37]  Brad J Marsh,et al.  Expedited approaches to whole cell electron tomography and organelle mark-up in situ in high-pressure frozen pancreatic islets. , 2008, Journal of structural biology.

[38]  P. Gilon,et al.  Glucose Acutely Decreases pH of Secretory Granules in Mouse Pancreatic Islets , 2006, Journal of Biological Chemistry.

[39]  Gerry McDermott,et al.  X-ray tomography of whole cells. , 2005, Current opinion in structural biology.

[40]  Jeffrey A. Fessler,et al.  Compensation for nonuniform resolution using penalized-likelihood reconstruction in space-variant imaging systems , 2004, IEEE Transactions on Medical Imaging.

[41]  S. Theander,et al.  Glucose sensitivity and metabolism-secretion coupling studied during two-year continuous culture in INS-1E insulinoma cells. , 2004, Endocrinology.

[42]  P. Rorsman,et al.  Insulin granule dynamics in pancreatic beta cells , 2003, Diabetologia.

[43]  G. Rutter,et al.  Involvement of conventional kinesin in glucose-stimulated secretory granule movements and exocytosis in clonal pancreatic β-cells , 2002, Journal of Cell Science.

[44]  C. Wollheim,et al.  Mitochondrial function in normal and diabetic β-cells , 2001, Nature.

[45]  B. Niemann,et al.  Tomographic imaging of biological specimens with the cryo transmission X-ray microscope , 2001 .

[46]  N. Porksen,et al.  Overnight inhibition of insulin secretion restores pulsatility and proinsulin/insulin ratio in type 2 diabetes. , 2000, American journal of physiology. Endocrinology and metabolism.

[47]  Erik Renström,et al.  The Cell Physiology of Biphasic Insulin Secretion. , 2000, News in physiological sciences : an international journal of physiology produced jointly by the International Union of Physiological Sciences and the American Physiological Society.

[48]  C. Wollheim,et al.  Beta-cell mitochondria in the regulation of insulin secretion: a new culprit in Type II diabetes , 2000, Diabetologia.

[49]  M. Noda,et al.  Identification of the docked granule pool responsible for the first phase of glucose-stimulated insulin secretion. , 1999, Diabetes.

[50]  T. Miki,et al.  The structure and function of the ATP-sensitive K+ channel in insulin-secreting pancreatic beta-cells. , 1999, Journal of molecular endocrinology.

[51]  L. Eliasson,et al.  Rapid ATP‐Dependent Priming of Secretory Granules Precedes Ca2+ ‐Induced Exocytosis in Mouse Pancreatic B‐Cells , 1997, The Journal of physiology.

[52]  I. Llona.  Synaptic like microvesicles : Do they participate in regulated exocytosis? , 1995, Neurochemistry International.

[53]  P. De Camilli,et al.  GABA and pancreatic beta‐cells: colocalization of glutamic acid decarboxylase (GAD) and GABA with synaptic‐like microvesicles suggests their role in GABA storage and secretion. , 1991, The EMBO journal.

[54]  P. Lacy,et al.  ISOLATION AND PROPERTIES OF SECRETORY GRANULES FROM RAT ISLETS OF LANGERHANS , 1969, The Journal of cell biology.

[55]  A. Bhattacharyya.  On a measure of divergence between two statistical populations defined by their probability distributions , 1943 .

[56]  Beer.  Bestimmung der Absorption des rothen Lichts in farbigen Flüssigkeiten , 1852 .