Topography of signaling molecules as detected by electron microscopy on plasma membrane sheets isolated from non-adherent mast cells
Jazyk angličtina Země Nizozemsko Médium print-electronic
Typ dokumentu srovnávací studie, časopisecké články, práce podpořená grantem
PubMed
17900607
DOI
10.1016/j.jim.2007.08.015
PII: S0022-1759(07)00284-0
Knihovny.cz E-zdroje
- MeSH
- buněčná membrána ultrastruktura MeSH
- imunoblotting MeSH
- klinické laboratorní techniky * MeSH
- krysa rodu Rattus MeSH
- mastocyty imunologie ultrastruktura MeSH
- membránové proteiny imunologie ultrastruktura MeSH
- myši MeSH
- průtoková cytometrie MeSH
- receptory IgE imunologie ultrastruktura MeSH
- transmisní elektronová mikroskopie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- Názvy látek
- membránové proteiny MeSH
- receptory IgE MeSH
Immunolabeling of isolated plasma membrane (PM) sheets combined with high-resolution electron microscopy is a powerful technique for understanding the topography of PM-bound signaling molecules. However, this technique has been mostly confined to analysis of membrane sheets from adherent cells. Here we present a rapid, simple and versatile method for isolation of PM sheets from non-adherent cells, and show its use for examination of the topography of Fcepsilon receptor I (FcepsilonRI) and transmembrane adaptors, LAT (linker for activation of T cells) and NTAL (non-T cell activation linker), in murine bone marrow-derived mast cells (BMMC). The data were compared with those obtained from widely used but tumor-derived rat basophilic leukemia (RBL) cells. In non-activated cells, FcepsilonRI was distributed either individually or in small clusters of comparable size in both cell types. In multivalent antigen-activated BMMC as well as RBL cells, FcepsilonRI was internalized to a similar extent, but, strikingly, internalization in BMMC was not preceded by formation of large (~200 nm) aggregates of FcepsilonRI, described previously in activated RBL cells. On the other hand, downstream adaptor proteins, LAT and NTAL, were localized in independent domains in both BMMC and RBL cells before and after FcepsilonRI triggering. The combined data demonstrate unexpected properties of FcepsilonRI signaling assemblies in BMMC and emphasize the importance of studies of PM sheets isolated from non-tumor cells.
Citace poskytuje Crossref.org
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