Characterizing relationships between cell structures and functions requires quantitative mesoscale mapping of intact cells showing subcellular rearrangements following stimulation. Thus, we investigated the application of soft X-ray tomography (SXT) as an approach for quantitative structural cell biology. Using SXT we generated three-dimensional reconstructions of whole pancreatic β-cells at different timepoints following glucose-stimulated insulin secretion. Reconstructions following stimulation show distinct insulin vesicle distribution patterns reflective of altered vesicle pool sizes as they mature and travel through the secretory pathway. Our results show that glucose stimulation causes rapid changes in biochemical composition and/or density of insulin packing, increased mitochondrial volume, and closer proximity of insulin vesicles to mitochondria. We are interested in understanding how different protein signaling networks that impact insulin secretion also impact cellular organization. Thus, we are screening the impact of multiple chemical probes on β-cell reoragnization and molecular maturation of insulin vesicles. This approach provides unique perspectives on the coordinated structural reorganization and interactions of organelles that dictate cell responses. Quantatitive structural cell biology appraoches will add new characterization of the interplay between cell structure and function during complex biological process and the progression of disease.

Quantitative structural mapping of the insulin secretory response

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