Nanoscopic spine localization of Norbin, an mGluR5 accessory protein
2014 (English)In: BMC neuroscience (Online), ISSN 1471-2202, Vol. 15, 45- p.Article in journal (Refereed) Published
Background: Norbin is a neuron-specific, cytosolic protein that interacts with the metabotropic glutamate receptor 5 (mGluR5) and has a profound impact on mGluR5 signaling. Yet, little is known about its synaptic distribution. Results: Here we have analyzed the spatial relationship between Norbin, postsynaptic density protein 95 (PSD-95), actin and mGluR5 in spines using super-resolution microscopy. Norbin was found to have a high degree of colocalization with actin and a lower degree of colocalization with PSD-95. Co-immunoprecipitation studies confirmed that interaction occurs between Norbin and actin, but not between Norbin and PSD-95. Norbin was also found to have a high degree of colocalization with the perisynaptically located mGluR5. Findings based on structured illumination microscopy (3D-SIM) of exogenous expressed Norbin-GFP were confirmed by stimulated emission depletion microscopy (STED) of immunolabeled endogenous Norbin. Conclusions: Norbin associates with actin rather than with PSD-95 in dendritic spines. Results regarding protein localization and colocalization performed with conventional confocal microscopy must be interpreted with great caution. The now available super-resolution microscopy techniques provide more accurate information about sub-cellular protein localization than previously was possible.
Place, publisher, year, edition, pages
BioMed Central, 2014. Vol. 15, 45- p.
Super-resolution microscopy, Colocalization, Norbin, Dendritic spines
IdentifiersURN: urn:nbn:se:kth:diva-145606DOI: 10.1186/1471-2202-15-45ISI: 000334951100001ScopusID: 2-s2.0-84897525805OAI: oai:DiVA.org:kth-145606DiVA: diva2:719205
FunderSwedish Research CouncilScience for Life Laboratory - a national resource center for high-throughput molecular bioscience
QC 201405232014-05-232014-05-232014-05-23Bibliographically approved