Journal Article

DZNE-2020-07522

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Novel standards in the measurement of rat insulin granules combining electron microscopy, high-content image analysis and in silico modelling.

Fava, E. (First author)DZNE* ; Dehghany, J. ; Ouwendijk, J. ; Müller, A. ; Niederlein, A. ; Verkade, P. ; Meyer-Hermann, M. (Corresponding author) ; Solimena, M.

2012
Springer Heidelberg

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Please use a persistent id in citations: doi:10.1007/s00125-011-2438-4

Abstract: Knowledge of number, size and content of insulin secretory granules is pivotal for understanding the physiology of pancreatic beta cells. Here we re-evaluated key structural features of rat beta cells, including insulin granule size, number and distribution as well as cell size.Electron micrographs of rat beta cells fixed either chemically or by high-pressure freezing were compared using a high-content analysis approach. These data were used to develop three-dimensional in silico beta cell models, the slicing of which would reproduce the experimental datasets.As previously reported, chemically fixed insulin secretory granules appeared as hollow spheres with a mean diameter of ∼350 nm. Remarkably, most granules fixed by high-pressure freezing lacked the characteristic halo between the dense core and the limiting membrane and were smaller than their chemically fixed counterparts. Based on our analyses, we conclude that the mean diameter of rat insulin secretory granules is 243 nm, corresponding to a surface area of 0.19 μm(2). Rat beta cells have a mean volume of 763 μm(3) and contain 5,000-6,000 granules.A major reason for the lower mean granule number/rat beta cell relative to previous accounts is a reduced estimation of the mean beta cell volume. These findings imply that each granule contains about twofold more insulin, while its exocytosis increases membrane capacitance about twofold less than assumed previously. Our integrated approach defines new standards for quantitative image analysis of beta cells and could be applied to other cellular systems.

Keyword(s): Animals (MeSH) ; Cells, Cultured (MeSH) ; Cytoplasmic Granules: metabolism (MeSH) ; Cytoplasmic Granules: ultrastructure (MeSH) ; Female (MeSH) ; Insulin: metabolism (MeSH) ; Insulin-Secreting Cells: metabolism (MeSH) ; Insulin-Secreting Cells: ultrastructure (MeSH) ; Microscopy, Electron, Transmission (MeSH) ; Rats (MeSH) ; Rats, Wistar (MeSH) ; Secretory Vesicles: metabolism (MeSH) ; Secretory Vesicles: ultrastructure (MeSH) ; Insulin

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Contributing Institute(s):

1. Systems Phenomics (AG Fava 1)

Research Program(s):

1. 342 - Disease Mechanisms and Model Systems (POF3-342) (POF3-342)

Appears in the scientific report 2012

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Database coverage:
; ; ; BIOSIS Previews ; Clarivate Analytics Master Journal List ; Current Contents - Clinical Medicine ; Current Contents - Life Sciences ; IF >= 5 ; JCR ; NCBI Molecular Biology Database ; Nationallizenz ; SCOPUS ; Science Citation Index ; Science Citation Index Expanded ; Web of Science Core Collection

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