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000141200 037__ $$aDZNE-2020-07522
000141200 041__ $$aEnglish
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000141200 1001_ $$0P:(DE-2719)2159508$$aFava, E.$$b0$$eFirst author$$udzne
000141200 245__ $$aNovel standards in the measurement of rat insulin granules combining electron microscopy, high-content image analysis and in silico modelling.
000141200 260__ $$aHeidelberg$$bSpringer$$c2012
000141200 264_1 $$2Crossref$$3online$$bSpringer Science and Business Media LLC$$c2012-01-18
000141200 264_1 $$2Crossref$$3print$$bSpringer Science and Business Media LLC$$c2012-04-01
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000141200 520__ $$aKnowledge 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.
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000141200 650_7 $$2NLM Chemicals$$aInsulin
000141200 650_2 $$2MeSH$$aAnimals
000141200 650_2 $$2MeSH$$aCells, Cultured
000141200 650_2 $$2MeSH$$aCytoplasmic Granules: metabolism
000141200 650_2 $$2MeSH$$aCytoplasmic Granules: ultrastructure
000141200 650_2 $$2MeSH$$aFemale
000141200 650_2 $$2MeSH$$aInsulin: metabolism
000141200 650_2 $$2MeSH$$aInsulin-Secreting Cells: metabolism
000141200 650_2 $$2MeSH$$aInsulin-Secreting Cells: ultrastructure
000141200 650_2 $$2MeSH$$aMicroscopy, Electron, Transmission
000141200 650_2 $$2MeSH$$aRats
000141200 650_2 $$2MeSH$$aRats, Wistar
000141200 650_2 $$2MeSH$$aSecretory Vesicles: metabolism
000141200 650_2 $$2MeSH$$aSecretory Vesicles: ultrastructure
000141200 7001_ $$aDehghany, J.$$b1
000141200 7001_ $$aOuwendijk, J.$$b2
000141200 7001_ $$0P:(DE-HGF)0$$aMüller, A.$$b3
000141200 7001_ $$aNiederlein, A.$$b4
000141200 7001_ $$aVerkade, P.$$b5
000141200 7001_ $$0P:(DE-HGF)0$$aMeyer-Hermann, M.$$b6$$eCorresponding author
000141200 7001_ $$aSolimena, M.$$b7
000141200 77318 $$2Crossref$$3journal-article$$a10.1007/s00125-011-2438-4$$b : Springer Science and Business Media LLC, 2012-01-18$$n4$$p1013-1023$$tDiabetologia$$v55$$x0012-186X$$y2012
000141200 773__ $$0PERI:(DE-600)1458993-X$$a10.1007/s00125-011-2438-4$$gVol. 55, no. 4, p. 1013 - 1023$$n4$$p1013-1023$$q55:4<1013 - 1023$$tDiabetologia$$v55$$x0012-186X$$y2012
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