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

Abstract

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.

Authors:

White, Kate L.  ^[1];

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• Search DOE PAGES for ORCID "0000-0001-8894-9621"

Singla, Jitin  ^[2];

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• Search DOE PAGES for ORCID "0000-0001-6225-0783"

Loconte, Valentina  ^[3];

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• Search DOE PAGES for ORCID "0000-0001-5715-9993"

Chen, Jian-Hua  ^[4];

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• Search DOE PAGES for ORCID "0000-0002-7998-0878"

Ekman, Axel ^[5]; Sun, Liping ^[3]; Zhang, Xianjun  ^[6];

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• Search DOE PAGES for ORCID "0000-0003-3554-780X"

Francis, John Paul ^[7]; Li, Angdi  ^[3];

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• Search DOE PAGES for ORCID "0000-0001-6143-6295"

Lin, Wen ^[8]; Tseng, Kaylee  ^[6];

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• Search DOE PAGES for ORCID "0000-0001-7450-2247"

McDermott, Gerry ^[4]; Alber, Frank  ^[2];

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Sali, Andrej  ^[9];

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• Search DOE PAGES for ORCID "0000-0003-0435-6197"

Larabell, Carolyn  ^[4];

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• Search DOE PAGES for ORCID "0000-0002-6262-4789"

Stevens, Raymond C.  ^[10]

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• Search DOE PAGES for ORCID "0000-0002-4522-8725"

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+ Show Author Affiliations

1. Department of Biological Sciences, Bridge Institute, USC Michelson Center for Convergent Bioscience, University of Southern California, Los Angeles, CA 90089, USA., Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.
2. Department of Biological Sciences, Bridge Institute, USC Michelson Center for Convergent Bioscience, University of Southern California, Los Angeles, CA 90089, USA., Institute for Quantitative and Computational Biosciences, Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90095, USA.
3. iHuman Institute, School of Life Science and Technology, ShanghaiTech University, Shanghai 201210, China.
4. Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA., Department of Anatomy, University of California, San Francisco, San Francisco, CA 94143, USA.
5. Department of Anatomy, University of California, San Francisco, San Francisco, CA 94143, USA.
6. Department of Biological Sciences, Bridge Institute, USC Michelson Center for Convergent Bioscience, University of Southern California, Los Angeles, CA 90089, USA.
7. Department of Computer Science, Bridge Institute, USC Michelson Center for Convergent Bioscience, University of Southern California, Los Angeles, CA 90089, USA.
8. Department of Chemistry, Bridge Institute, USC Michelson Center for Convergent Bioscience, University of Southern California, Los Angeles, CA 90089, USA.
9. California Institute for Quantitative Biosciences, Department of Bioengineering and Therapeutic Sciences, Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94158, USA.
10. Department of Biological Sciences, Bridge Institute, USC Michelson Center for Convergent Bioscience, University of Southern California, Los Angeles, CA 90089, USA., iHuman Institute, School of Life Science and Technology, ShanghaiTech University, Shanghai 201210, China., Department of Chemistry, Bridge Institute, USC Michelson Center for Convergent Bioscience, University of Southern California, Los Angeles, CA 90089, USA.

Publication Date:

Wed Dec 09 00:00:00 EST 2020

Research Org.:

Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Org.:

USDOE Office of Science (SC), Biological and Environmental Research (BER); National Institutes of Health (NIH); National Natural Science Foundation of China (NSFC)

OSTI Identifier:

1734835

Alternate Identifier(s):

OSTI ID: 1763702

Grant/Contract Number:  

AC02-05CH11231; P41GM103445; R01GM083960; P41GM109824; 31950410543

Resource Type:

Published Article

Journal Name:

Science Advances

Additional Journal Information:

Journal Name: Science Advances Journal Volume: 6 Journal Issue: 50; Journal ID: ISSN 2375-2548

Publisher:

AAAS

Country of Publication:

United States

Language:

English

Subject:

59 BASIC BIOLOGICAL SCIENCES