Structure of cytomatrix and nuclear matrix revealed by embedment-free electron microscopy

• Autorzy: Gajkowska B , Cholewinski M. , Gniadecki R.
• Języki publikacji: EN

Abstrakty

EN

Embedment-free electron microscopy (EFEM) is a new method which allows the visualisation of cytoskeleton in whole-mounted cells. In this study we employed EFEM to investigate the structure of cellular scaffolds in glioma C6 cell line. The cells were extracted with Triton X-100 that dissolves phospholipids in the membranes and removes most of cytoplasmic soluble proteins. The DNA and nuclear histones were removed with DNase I and high-salt buffer, respectively. The remaining cellular frameworks were temporary embedded in diethylene glycol distearate (DGD), sectioned and observed in transmission and scanning electron microscope after the removal of DGD. The predominant structure was the extensive meshwork of 10-20 nm filaments in the cytoplasm (cytomatrix) and 15-30 nm filaments in the nucleus (nuclear matrix). The 5 nm filaments, presumably corresponding to the actin filaments, were present in the cytomatrix, but not in the nuclear matrix. Moreover, the ultrathin (3 nm) filaments, connecting other cytoskeletal components were detected. Those are possibly identical with the previously described plectin filaments. For the first time we report the occurrence of ultrathin filaments in the nuclear matrix. Thus, in a addition to the well known cytoskeletal components (microtubules, intermediate filaments, actin microfilaments) EFEM showed a new type of filaments (the ultrathin filaments) in the cytomatrix and nuclear matrix. Further immunocytochemical studies are needed to determine the biochemical identity of the filaments observed in EFEM.

Słowa kluczowe

EN

embedment-free electron microscopy; cytoskeleton; cell extraction; cytomatrix; glioma; cell culture; phospholipid; fixation; immunocytochemical technique; membrane; filament; microtubule; visualization; nuclear matrix; DNA; microfilament

• Wydawca: -
• Czasopismo: Acta Neurobiologiae Experimentalis
• Rocznik: 2000
• Tom: 60
• Numer: 2
• Opis fizyczny: p.147-158,fig.

Twórcy

autor

Gajkowska B

• Polish Academy of Sciences, 5 Pawinski St., 02-106 Warsaw, Poland

autor

Cholewinski M.

autor

Gniadecki R.

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