Correlative Light and Electron Microscopy Reveals the HAS3-Induced Dorsal Plasma Membrane Ruffles

Hyaluronan is a linear sugar polymer synthesized by three isoforms of hyaluronan synthases (HAS1, 2, and 3) that forms a hydrated scaffold around cells and is an essential component of the extracellular matrix. The morphological changes of cells induced by active hyaluronan synthesis are well recogn...

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Main Authors: Kirsi Rilla, Arto Koistinen
Format: Article
Language:English
Published: Hindawi Limited 2015-01-01
Series:International Journal of Cell Biology
Online Access:http://dx.doi.org/10.1155/2015/769163
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spelling doaj-582dc4e85bac4ba9a37d687091a3b9082020-11-24T22:57:23ZengHindawi LimitedInternational Journal of Cell Biology1687-88761687-88842015-01-01201510.1155/2015/769163769163Correlative Light and Electron Microscopy Reveals the HAS3-Induced Dorsal Plasma Membrane RufflesKirsi Rilla0Arto Koistinen1Institute of Biomedicine and SIB Labs, University of Eastern Finland, 70211 Kuopio, FinlandInstitute of Biomedicine and SIB Labs, University of Eastern Finland, 70211 Kuopio, FinlandHyaluronan is a linear sugar polymer synthesized by three isoforms of hyaluronan synthases (HAS1, 2, and 3) that forms a hydrated scaffold around cells and is an essential component of the extracellular matrix. The morphological changes of cells induced by active hyaluronan synthesis are well recognized but not studied in detail with high resolution before. We have previously found that overexpression of HAS3 induces growth of long plasma membrane protrusions that act as platforms for hyaluronan synthesis. The study of these thin and fragile protrusions is challenging, and they are difficult to preserve by fixation unless they are adherent to the substrate. Thus their structure and regulation are still partly unclear despite careful imaging with different microscopic methods in several cell types. In this study, correlative light and electron microscopy (CLEM) was utilized to correlate the GFP-HAS3 signal and the surface ultrastructure of cells in order to study in detail the morphological changes induced by HAS3 overexpression. Surprisingly, this method revealed that GFP-HAS3 not only localizes to ruffles but in fact induces dorsal ruffle formation. Dorsal ruffles regulate diverse cellular functions, such as motility, regulation of glucose metabolism, spreading, adhesion, and matrix degradation, the same functions driven by active hyaluronan synthesis.http://dx.doi.org/10.1155/2015/769163
collection DOAJ
language English
format Article
sources DOAJ
author Kirsi Rilla
Arto Koistinen
spellingShingle Kirsi Rilla
Arto Koistinen
Correlative Light and Electron Microscopy Reveals the HAS3-Induced Dorsal Plasma Membrane Ruffles
International Journal of Cell Biology
author_facet Kirsi Rilla
Arto Koistinen
author_sort Kirsi Rilla
title Correlative Light and Electron Microscopy Reveals the HAS3-Induced Dorsal Plasma Membrane Ruffles
title_short Correlative Light and Electron Microscopy Reveals the HAS3-Induced Dorsal Plasma Membrane Ruffles
title_full Correlative Light and Electron Microscopy Reveals the HAS3-Induced Dorsal Plasma Membrane Ruffles
title_fullStr Correlative Light and Electron Microscopy Reveals the HAS3-Induced Dorsal Plasma Membrane Ruffles
title_full_unstemmed Correlative Light and Electron Microscopy Reveals the HAS3-Induced Dorsal Plasma Membrane Ruffles
title_sort correlative light and electron microscopy reveals the has3-induced dorsal plasma membrane ruffles
publisher Hindawi Limited
series International Journal of Cell Biology
issn 1687-8876
1687-8884
publishDate 2015-01-01
description Hyaluronan is a linear sugar polymer synthesized by three isoforms of hyaluronan synthases (HAS1, 2, and 3) that forms a hydrated scaffold around cells and is an essential component of the extracellular matrix. The morphological changes of cells induced by active hyaluronan synthesis are well recognized but not studied in detail with high resolution before. We have previously found that overexpression of HAS3 induces growth of long plasma membrane protrusions that act as platforms for hyaluronan synthesis. The study of these thin and fragile protrusions is challenging, and they are difficult to preserve by fixation unless they are adherent to the substrate. Thus their structure and regulation are still partly unclear despite careful imaging with different microscopic methods in several cell types. In this study, correlative light and electron microscopy (CLEM) was utilized to correlate the GFP-HAS3 signal and the surface ultrastructure of cells in order to study in detail the morphological changes induced by HAS3 overexpression. Surprisingly, this method revealed that GFP-HAS3 not only localizes to ruffles but in fact induces dorsal ruffle formation. Dorsal ruffles regulate diverse cellular functions, such as motility, regulation of glucose metabolism, spreading, adhesion, and matrix degradation, the same functions driven by active hyaluronan synthesis.
url http://dx.doi.org/10.1155/2015/769163
work_keys_str_mv AT kirsirilla correlativelightandelectronmicroscopyrevealsthehas3induceddorsalplasmamembraneruffles
AT artokoistinen correlativelightandelectronmicroscopyrevealsthehas3induceddorsalplasmamembraneruffles
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