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Spatial distribution of Na+-K+-ATPase in dendritic spines dissected by nanoscale superresolution STED microscopy
KTH, Skolan för teknikvetenskap (SCI), Tillämpad fysik, Experimentell biomolekylär fysik.ORCID-id: 0000-0002-5584-9170
KTH, Skolan för teknikvetenskap (SCI), Tillämpad fysik, Experimentell biomolekylär fysik.
Vise andre og tillknytning
2011 (engelsk)Inngår i: BMC neuroscience (Online), ISSN 1471-2202, E-ISSN 1471-2202, Vol. 12, s. 16-Artikkel i tidsskrift (Fagfellevurdert) Published
Abstract [en]

Background: The Na+,K+-ATPase plays an important role for ion homeostasis in virtually all mammalian cells, including neurons. Despite this, there is as yet little known about the isoform specific distribution in neurons. Results: With help of superresolving stimulated emission depletion microscopy the spatial distribution of Na+,K+-ATPase in dendritic spines of cultured striatum neurons have been dissected. The found compartmentalized distribution provides a strong evidence for the confinement of neuronal Na+,K+-ATPase (alpha 3 isoform) in the postsynaptic region of the spine. Conclusions: A compartmentalized distribution may have implications for the generation of local sodium gradients within the spine and for the structural and functional interaction between the sodium pump and other synaptic proteins. Superresolution microscopy has thus opened up a new perspective to elucidate the nature of the physiological function, regulation and signaling role of Na+,K+-ATPase from its topological distribution in dendritic spines.

sted, utgiver, år, opplag, sider
2011. Vol. 12, s. 16-
HSV kategori
Identifikatorer
URN: urn:nbn:se:kth:diva-31019DOI: 10.1186/1471-2202-12-16ISI: 000287447600001PubMedID: 21272290Scopus ID: 2-s2.0-79251589617OAI: oai:DiVA.org:kth-31019DiVA, id: diva2:402231
Forskningsfinansiär
Swedish Research Council, VR-2006-3197, VR-2007-4582, VR-2007-2519EU, FP7, Seventh Framework Programme, Fluodiamon 201 837Science for Life Laboratory - a national resource center for high-throughput molecular bioscience
Merknad

QC 20110307

Tilgjengelig fra: 2011-03-07 Laget: 2011-03-07 Sist oppdatert: 2018-01-12bibliografisk kontrollert
Inngår i avhandling
1. Super resolution optical imaging – image analysis, multicolor development and biological applications
Åpne denne publikasjonen i ny fane eller vindu >>Super resolution optical imaging – image analysis, multicolor development and biological applications
2014 (engelsk)Doktoravhandling, med artikler (Annet vitenskapelig)
Abstract [en]

This thesis focuses on super resolution STED optical imaging. STED provides a wealth of new informational content to the acquired images by using stimulated emission to surpass the diffraction limit in optical fluorescence microscopy. To further increase the informational content, a new method to perform multicolor STED imaging by exploiting differences in the photostability and excitation spectra of dyes is presented. In order to extract information from the images, computational algorithms which handle the new type of high resolution informational content are developed.

We propose that multicolor super resolution imaging in combination with image analysis can reduce the amount of clinical samples required to perform accurate cancer diagnosis. To date, such diagnosis is based mainly on significant amounts of tissue samples extracted from the suspected tumor site. The sample extraction often requires anesthetics and can lead to complications such as hematoma, infections and even cancer cell ceding along the needle track. We show that by applying multicolor STED and image analysis, the information gained from single cells is greatly increased. We therefore propose that accurate diagnosis can be based on significantly less extracted tissue material, allowing for a more patient friendly sampling. This approach can also be applied when studying blood platelets, where we show how the high informational content can be used to identify platelet specific activational states. Since platelets are involved in many different types of diseases, such analysis could provide means of performing truly minimally invasive diagnostics based on a simple blood test.

In addition, our data makes it possible to understand in finer detail the underlying mechanisms rendering cells metastasis competent. We combine the high resolution spatial information provided by STED with information regarding the adhesive forces of cells measured by TFM (Traction Force Microscopy) and the cell stiffness measured by AFM (Atomic Force Microscopy). Such comparisons provide a link between the specific highly resolved protein distributions and different cellular mechanics and functions.

This thesis also includes STED imaging and analysis on the spatial organization of neuronal synaptic regulating proteins, implicating the speed with which neuronal signaling can be regulated.

sted, utgiver, år, opplag, sider
Stockholm: KTH Royal Institute of Technology, 2014. s. x, 86
Serie
TRITA-FYS, ISSN 0280-316X ; 2014:04
Emneord
Stimulated emission depletion (STED) microscopy, nanoscopy, multicolor, image analysis, diagnostics, cancer, metastasis
HSV kategori
Forskningsprogram
Biologisk fysik
Identifikatorer
urn:nbn:se:kth:diva-141011 (URN)978-91-7595-001-3 (ISBN)
Disputas
2014-02-28, FB42, AlbaNova Universitetscentrum, Roslagstullsbacken 21, Stockholm, 10:51 (engelsk)
Opponent
Veileder
Merknad

QC 20140207

Tilgjengelig fra: 2014-02-07 Laget: 2014-02-05 Sist oppdatert: 2014-02-07bibliografisk kontrollert

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