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

Kate L White1,2, Jitin Singla3,4, Valentina Loconte5

  • 1Department of Biological Sciences, Bridge Institute, USC Michelson Center for Convergent Bioscience, University of Southern California, Los Angeles, CA 90089, USA. stevens@usc.edu calarabell@lbl.gov sali@salilab.org katewhit@usc.edu.

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Soft x-ray tomography reveals dynamic changes in pancreatic beta cells during insulin secretion. Glucose stimulation alters insulin vesicle distribution, mitochondrial volume, and vesicle-mitochondria proximity, offering new insights into cellular function.

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Area of Science:

  • Cell Biology
  • Endocrinology
  • Biophysics

Background:

  • Understanding cell structure-function relationships requires mesoscale mapping of intact cells.
  • Current imaging techniques have limitations in visualizing subcellular rearrangements post-stimulation.

Purpose of the Study:

  • To apply soft x-ray tomography for 3D reconstruction of pancreatic beta cells.
  • To investigate structural changes during glucose-stimulated insulin secretion.

Main Methods:

  • Soft x-ray tomography was used to generate 3D reconstructions of pancreatic beta cells.
  • Cells were analyzed at various time points after glucose stimulation.

Main Results:

  • Glucose stimulation altered insulin vesicle distribution and pool sizes within the secretory pathway.
  • Observed rapid changes in insulin packing, increased mitochondrial volume, and closer vesicle-mitochondria proximity.
  • Exendin-4 costimulation prolonged these effects and enhanced insulin packaging and vesicle maturation.

Conclusions:

  • Soft x-ray tomography provides novel insights into pancreatic beta cell structural dynamics.
  • Coordinated organelle reorganization dictates cellular responses to stimulation.
  • Findings illuminate the mechanisms of insulin secretion and potential therapeutic targets.