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Cytoplasmic actomyosin fibrils after preservation with high pressure freezing

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Summary

The fine structure of the actomyosin system of Physarum polycephalum was investigated in vitrified specimens after applying a pressure of >2.1 kbar and freezing rates of 500 to 5,000° C/s. The frozen specimens were either freeze-substituted or freeze-fractured and compared with material processed according to conventional methods of freeze-etching preparation.

Artifactual alterations, as seen in the form of destroyed areas of the cytoplasm after chemical fixation, were not observed after freeze-substitution. However, small ice crystals formed by recrystallization within most of the cytoplasmic actomyosin fibrils prevented a fine structural analysis.

Such a destruction of the fibrillar fine structure was not found after freezeetching. In replicas of deep-etched objects 10 nm-thick filaments were localized, which could be conclusively identified as F-actin. The actin filaments are located randomly in the peripheral cytoplasm forming the cell cortex. By the process of parallel aggregation, the filaments can be differentiated to fibrils. Thick myosin filaments were not observed. However, structures resembling cross bridges between single actin filaments suggest the existence of oligomeric myosin.

The present investigation shows that, in addition to biomembranes, other cytoplasmic differentiations such as components of the groundplasm can be successfully demonstrated employing the deep-etching technique when the freezing methods are improved by avoiding freeze-protection pretreatments.

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Authors and Affiliations

  1. Institute for Cytology, University of Bonn, Bonn, Federal Republic of Germany

    Klaus-Velten Wolf,  Wilhelm Stockem & Karl-Ernst Wohlfarth-Bottermann

  2. Institute for Cell Biology, Eidgenössische Technische Hochschule Zürich, Zürich, Switzerland

    Hans Moor

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  1. Klaus-Velten Wolf
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  2. Wilhelm Stockem
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  3. Karl-Ernst Wohlfarth-Bottermann
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  4. Hans Moor
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Dedicated to Prof. Dr. R. Danneel on the occasion of his 80th birthday

The authors wish to thank Dr. R.L. Snipes (Giessen) for translating the manuscript

The investigation was supported by a Grant from the Deutsche Forschungsgemeinschaft

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Wolf, KV., Stockem, W., Wohlfarth-Bottermann, KE. et al. Cytoplasmic actomyosin fibrils after preservation with high pressure freezing. Cell Tissue Res. 217, 479–495 (1981). https://doi.org/10.1007/BF00219359

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