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Nanoparticle-Based Immunocytochemistry Reveals Microarchitecture of the Cell Nucleus
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SYSNO ASEP 0389005 Document Type C - Proceedings Paper (int. conf.) R&D Document Type The record was not marked in the RIV Title Nanoparticle-Based Immunocytochemistry Reveals Microarchitecture of the Cell Nucleus Author(s) Hozák, Pavel (UMG-J) RID, ORCID Source Title Beyond the Limit of Histochemistry - 14th INTERNATIONAL CONGRESS OF HISTOCHEMISTRY AND CYTOCHEMISTRY. - Kyoto : International Federation of Societies for Histochemistry and Cytochemistry, Japan Society of Histochemistry, 2012 Number of pages 1 s. Action 14th International Congress of Histochemistry and Cytochemistry Event date 26.08.2012-29.08.2012 VEvent location Kyoto Country JP - Japan Event type WRD Language eng - English Country JP - Japan Keywords PIP2 ; NMI ; cell nucleus Subject RIV EB - Genetics ; Molecular Biology R&D Projects GAP305/11/2232 GA ČR - Czech Science Foundation (CSF) LC545 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) LC06063 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) FRTI3588 GA MPO - Ministry of Industry and Trade (MPO) Institutional support UMG-J - RVO:68378050 CEZ AV0Z50520514 - UMG-J (2005-2011) Annotation I will summarize the current possibilities of TEM visualization of molecules in cells. In order to overcome the current limitations of immunodetection, we prepared a set of novel nanoparticles (NPs) which fulfill several criteria: size in the frame of 5-12 nm, small size distribution, good contrast and stability in the electron microscope, stability of colloidal solution during conjugation, and surface properties allowing for conjugation with antibodies With the use of novel NPs, various combinations with commercial gold NPs can be made to obtain a set for simultaneous labeling. For the first time in ultrastructural histochemistry, up to five molecular targets can be identified simultaneously. These methods allowed us to progress with understanding some novel molecular interactions in the cell nucleus. I will discuss interactions of phosphatidylinositol-4,5-bisphosphate (PIP2) and nuclear myosin I (NMI) which are involved in regulation of gene expression. PIP2 resides in the nucleus in a different form than the classical bilayer membrane, apparently forming specific nuclear protein complexes. Our data suggest that nucleolar PIP2 might serve as a transcription factor for ribosomal genes. We therefore investigated PIP distribution in cell nuclei with a special attention to nucleoli by ultrastructural tomography, and mapped PIP colocalization with various factors involved in RNA pol I transcription. We also showed in living cells that NM1 binds to PIP2 in the cell nucleus, and this was further confirmed by electron microscopy and molecular approaches. The results will be discussed in the frame of the current model of the nucleolus and lipid functions in the cell nucleus. Workplace Institute of Molecular Genetics Contact Nikol Škňouřilová, nikol.sknourilova@img.cas.cz, Tel.: 241 063 217 Year of Publishing 2014
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