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Cryoelectron Tomography: Implications for Actin Cytoskeleton Research

Croatian International Relations Review

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Title Cryoelectron Tomography: Implications for Actin Cytoskeleton Research
Krioelektronska tomografija: implikacije za istra`ivanje aktinskoga citoskeleta
 
Creator Weber, Igor
 
Subject actin; cytoskeleton; cryofixation; electron microscopy; tomography
 
Description Disclosing undistorted spatial organization of the actin cytoskeleton with molecular resolution is fundamental for understanding the cellular ultrastructure. Whereas important insights into the architecture of microfilament networks have been gained by negative staining, critical point drying, and freeze-fracturing methods, it is cryoelectron tomography that provides, for the first time, a comprehensive three-dimensional view of the intact actin cytoskeleton in situ. In particular, topological relationships such as microfilament three-dimensional proximity, angles at filament branching points, and modes of microfilament interaction with the plasma membrane can be visualized with an unprecedented accuracy using this technique. Further improvements are expected to bring the resolution into the realm of 2–3 nm, where automatic pattern- recognition methods can be applied to identify actin-binding complexes. Combining cryoelectron tomography with ultra-fine immunolabeling and high-resolution fluorescence microscopy will make it possible to correlate structural data on the nanometer scale with molecular specificity and dynamical information.
Razotkrivanje netaknute prostorne organizacije aktinskoga citoskeleta na molekularnoj razini od temeljne je važnosti za razumijevanje stanične ultrastrukture. Iako su tehnike negativnoga bojanja, sušenja na kritičnoj točki i kalanja smrznutih uzoraka omogućile značajne uvide u arhitekturu umrežavanja mikrofilamenata, tek je krioelektronska tomografija po prvi puta otvorila sveobuhvatni trodimenzionalni vidik na intaktni aktinski citoskelet in situ. Posebice, topološki odnosi poput me|usobnoga položaja mikrofilamenata u prostoru, kutova grananja filamenata, kao i načina interakcije filamenata sa staničnom membranom mogu biti predočeni pomoću ove tehnike s do sada nedosegnutom točnošću. Očekuje se daljnje poboljšanje moći razlučivanja do područja izme|u 2 i 3 nm, kada će biti moguće primijeniti automatske metode prepoznavanja uzoraka za identifikaciju kompleksa vezanih uz aktin. Kombiniranje krioelektronske tomografije s ultrafinim imunooznačavanjem i fluorescencijskom mikroskopijom velike rezolucije omogućit će korelaciju strukturnih podataka na nanometarskoj skali s molekularnom specifičnošću i dinamičkom informacijom.
 
Publisher Croatian Chemical Society
 
Date 2005-09-15
 
Type text
info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
 
Format application/pdf
 
Identifier http://hrcak.srce.hr/37
http://hrcak.srce.hr/file/37
 
Source Croatica chemica acta; Vol.78 No.3; ISSN 0011-1643 (Print); ISSN 1334-417X (Online)
 
Language en
 
Rights info:eu-repo/semantics/openAccess
Parts of the contents of Croat. Chem. Acta (e. g. figures or tables) may be reproduced without prior permission, provided reference is made to their source.