3D ultrastructural analysis of α‐granule, dense granule, mitochondria, and canalicular system arrangement in resting human platelets

Abstract Background State‐of‐the‐art 3‐dimensional (3D) electron microscopy approaches provide a new standard for the visualization of human platelet ultrastructure. Application of these approaches to platelets rapidly fixed prior to purification to minimize activation should provide new insights in...

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Autores principales:	Irina D. Pokrovskaya, Shilpi Yadav, Amith Rao, Emma McBride, Jeffrey A. Kamykowski, Guofeng Zhang, Maria A. Aronova, Richard D. Leapman, Brian Storrie
Formato:	article
Lenguaje:	EN
Publicado:	Wiley 2020
Materias:	electron microscopy hemostasis platelet granules platelet organelles platelet ultrastructure super‐resolution light microscopy Diseases of the blood and blood-forming organs RC633-647.5
Acceso en línea:	https://doaj.org/article/9e2820b32b544c35b0587580e299144e
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spelling	oai:doaj.org-article:9e2820b32b544c35b0587580e299144e2021-11-15T06:10:44Z3D ultrastructural analysis of α‐granule, dense granule, mitochondria, and canalicular system arrangement in resting human platelets2475-037910.1002/rth2.12260https://doaj.org/article/9e2820b32b544c35b0587580e299144e2020-01-01T00:00:00Zhttps://doi.org/10.1002/rth2.12260https://doaj.org/toc/2475-0379Abstract Background State‐of‐the‐art 3‐dimensional (3D) electron microscopy approaches provide a new standard for the visualization of human platelet ultrastructure. Application of these approaches to platelets rapidly fixed prior to purification to minimize activation should provide new insights into resting platelet ultrastructure. Objectives Our goal was to determine the 3D organization of α‐granules, dense granules, mitochondria, and canalicular system in resting human platelets and map their spatial relationships. Methods We used serial block face–scanning electron microscopy images to render the 3D ultrastructure of α‐granules, dense granules, mitochondria, canalicular system, and plasma membrane for 30 human platelets, 10 each from 3 donors. α‐Granule compositional data were assessed by sequential, serial section cryo‐immunogold electron microscopy and by immunofluorescence (structured illumination microscopy). Results and Conclusions α‐Granule number correlated linearly with platelet size, while dense granule and mitochondria number had little correlation with platelet size. For all subcellular compartments, individual organelle parameters varied considerably and organelle volume fraction had little correlation with platelet size. Three‐dimensional data from 30 platelets indicated only limited spatial intermixing of the different organelle classes. Interestingly, almost 70% of α‐granules came within ≤35 nm of each other, a distance associated in other cell systems with protein‐mediated contact sites. Size and shape analysis of the 1488 α‐granules analyzed revealed no more variation than that expected for a Gaussian distribution. Protein distribution data indicated that all α‐granules likely contained the same major set of proteins, albeit at varying amounts and varying distribution within the granule matrix.Irina D. PokrovskayaShilpi YadavAmith RaoEmma McBrideJeffrey A. KamykowskiGuofeng ZhangMaria A. AronovaRichard D. LeapmanBrian StorrieWileyarticleelectron microscopyhemostasisplatelet granulesplatelet organellesplatelet ultrastructuresuper‐resolution light microscopyDiseases of the blood and blood-forming organsRC633-647.5ENResearch and Practice in Thrombosis and Haemostasis, Vol 4, Iss 1, Pp 72-85 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	electron microscopy hemostasis platelet granules platelet organelles platelet ultrastructure super‐resolution light microscopy Diseases of the blood and blood-forming organs RC633-647.5
spellingShingle	electron microscopy hemostasis platelet granules platelet organelles platelet ultrastructure super‐resolution light microscopy Diseases of the blood and blood-forming organs RC633-647.5 Irina D. Pokrovskaya Shilpi Yadav Amith Rao Emma McBride Jeffrey A. Kamykowski Guofeng Zhang Maria A. Aronova Richard D. Leapman Brian Storrie 3D ultrastructural analysis of α‐granule, dense granule, mitochondria, and canalicular system arrangement in resting human platelets
description	Abstract Background State‐of‐the‐art 3‐dimensional (3D) electron microscopy approaches provide a new standard for the visualization of human platelet ultrastructure. Application of these approaches to platelets rapidly fixed prior to purification to minimize activation should provide new insights into resting platelet ultrastructure. Objectives Our goal was to determine the 3D organization of α‐granules, dense granules, mitochondria, and canalicular system in resting human platelets and map their spatial relationships. Methods We used serial block face–scanning electron microscopy images to render the 3D ultrastructure of α‐granules, dense granules, mitochondria, canalicular system, and plasma membrane for 30 human platelets, 10 each from 3 donors. α‐Granule compositional data were assessed by sequential, serial section cryo‐immunogold electron microscopy and by immunofluorescence (structured illumination microscopy). Results and Conclusions α‐Granule number correlated linearly with platelet size, while dense granule and mitochondria number had little correlation with platelet size. For all subcellular compartments, individual organelle parameters varied considerably and organelle volume fraction had little correlation with platelet size. Three‐dimensional data from 30 platelets indicated only limited spatial intermixing of the different organelle classes. Interestingly, almost 70% of α‐granules came within ≤35 nm of each other, a distance associated in other cell systems with protein‐mediated contact sites. Size and shape analysis of the 1488 α‐granules analyzed revealed no more variation than that expected for a Gaussian distribution. Protein distribution data indicated that all α‐granules likely contained the same major set of proteins, albeit at varying amounts and varying distribution within the granule matrix.
format	article
author	Irina D. Pokrovskaya Shilpi Yadav Amith Rao Emma McBride Jeffrey A. Kamykowski Guofeng Zhang Maria A. Aronova Richard D. Leapman Brian Storrie
author_facet	Irina D. Pokrovskaya Shilpi Yadav Amith Rao Emma McBride Jeffrey A. Kamykowski Guofeng Zhang Maria A. Aronova Richard D. Leapman Brian Storrie
author_sort	Irina D. Pokrovskaya
title	3D ultrastructural analysis of α‐granule, dense granule, mitochondria, and canalicular system arrangement in resting human platelets
title_short	3D ultrastructural analysis of α‐granule, dense granule, mitochondria, and canalicular system arrangement in resting human platelets
title_full	3D ultrastructural analysis of α‐granule, dense granule, mitochondria, and canalicular system arrangement in resting human platelets
title_fullStr	3D ultrastructural analysis of α‐granule, dense granule, mitochondria, and canalicular system arrangement in resting human platelets
title_full_unstemmed	3D ultrastructural analysis of α‐granule, dense granule, mitochondria, and canalicular system arrangement in resting human platelets
title_sort	3d ultrastructural analysis of α‐granule, dense granule, mitochondria, and canalicular system arrangement in resting human platelets
publisher	Wiley
publishDate	2020
url	https://doaj.org/article/9e2820b32b544c35b0587580e299144e
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3D ultrastructural analysis of α‐granule, dense granule, mitochondria, and canalicular system arrangement in resting human platelets

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