PubMed Health⌕ Search

Biomedical subjects

I K Buckley

Publications and source records attributed to I K Buckley.

16 recordsLinked to original sources
PubMed1981

Claims that intermediate filaments contain F-actin are unwarranted.

Examination of sectioned, embedded material showed that almost all intermediate filaments normally present in cultured chick embryo fibroblasts, IMR-33 cells, and BHK-21 cells were eliminated by trypsinization. At the same time atypical filaments of intermediate size were formed by close apposition of thin filaments. Previous observations (Buckley et al., 1978, J. Cell Biol. 78:644-652) on negatively stained and on critical-point-dried whole-cell mounts confused these different filament types and, on the basis of heavy meromyosin binding to the atypical intermediate-sized filaments, we suggested that many normal intermediate filaments contain f-actin. The present results show that this suggestion was unwarranted and we therefore retract it.

Actins↗
PubMed1978

Heavy meromyosin labeling of intermediate filaments in cultured connective tissue cells.

Mild treatment with trypsin causes a radical change in the heavy meromyosin (HMM) binding properties of intermediate filaments in glycerinated, myosin-extracted cultured chick embryo connective tissue cells. In non-trypsin-treated cells, HMM labeling of filaments was often indistinct and variable in its distribution. By contrast, in cells treated with trypsin (under conditions which allowed most intermediate filaments to survive), virtually all filaments, including those of intermediate size, decorated with HMM to give distinct arrowhead patterns. We suggest that most intermediate filaments in such cells contain a core of F-actin masked by trypsin-labile accessory proteins.

Actins↗
PubMed1976

Form and distribution of actin and myosin in non-muscle cells: a study using cultured chick embryo fibroblasts.

Attempting to throw light on the mechanical basis of movement of non-muscle (cf. muscle) cells, the present work aims to determine the form and distribution of actin and myosin in chick embryo fibroblasts. These cells were cultured on formvar, fixed in glutaraldehyde then osmium tetroxide vapours, dehydrated, critical-point dried and examined, in toto, in the electron microscope (EM). Stereoscopic pairs of micrographs were studied to define more exactly the form and distribution of cytoplasmic filaments topographically associated with deformations of the cell surface and with organelle movements through the cytoplasm. Permeating the cytoplasm, interconnecting long and short filaments closely surrounded all organelles, linked with microtubules and polyribosomes and joined to the plasma membrane. These filaments, which varied greatly in width (2-13 nm) were closely associated with large numbers of 'comma-shaped' globoid bodies of approximately 15 nm diameter. Attempting to establish the identity, form and distribution of cytoplasmic myosin, cultured cells were extracted with a cold (4 degrees C) glycerol/pyrophosphate solution for 24 h before being fixed and critical-point dried. EM examination of these cells revealed a residual three-dimensional network of branching and anastomosing 4-13 nm diameter smooth filaments, devoid of fine (2 nm) filaments and globoid bodies. Examination of fixed, critical-point dried, skeletal muscle heavy meromyosin showed globoid structures similar in form and size to the globoid bodies found in cultures fibroblasts. Similarly fixed and critical-point dried paracrystals of actin, polymerized in the presence of Mg2+, appeared as branching interconnecting filaments which, in form and dimensions, resembled the network filaments observed in pyrophosphate-extracted cells. It is concluded that the pyrophosphate-extractable globoid bodies found in cultured fibroblasts represent monomers of myosin, that the broader filaments to which these attach represent actin in Mg2+ paracrystalline form and that the various subcellular movements are brought about by interactions between the two, analogous to those occurring in muscle cells.

Actins↗
PubMed1975

Cell models in the study of mechanisms of toxicity.

Cell models, particularly in vitro systems, allow studies of toxicological mechanisms to be made under controlled conditions. One way that such mechanisms can be studied is by determining the reproducible changes that toxins induce, at the electron microscopic level, in the structure of cultured cells. A detailed description of such a study is given and includes preparation of the cell model, treatment of the cells, assessment of toxin-induced changes, and some general conclusions derived from experiments with this cell model. Other models which have been used for toxicological studies are also discussed with reference to advantages, limitations and potential applications.

Adult↗
PubMed1975

Three dimensional fine structure of cultured cells: possible implications for subcellular motility.

To determine the three dimensional fine structure of whole motile cells, rat embryo cells, cultured on Formvar-coated cover-glasses, were glutaraldehyde/osmium-fixed, mounted on grids, dehydrated, critical point dried and examined by transmission electron microscopy using stereoscopic techniques. Three dimensional arrays of organelles occurred in a filament-rich cytoplasmic matrix. Here, besides microtubules and elongate filaments, inter-connected filaments formed a widespread fine-mesh space network which attached to the plasma membrane and closely surrounded all organelles. Negative staining revealed a similar newtork in unfixed cells. It is concluded that this network represents part of the force-generating mechanism for various subcellular movements.

Animals↗
PubMed1975

Electron microscopy of critical point dried whole cultured cells.

To determine the overall fine structure of whole, unsectioned cells, cells from rat embryos were cultured on Formvar, glutaraldehyde/osmium-fixed, transferred to grids, dehypdrated, critical point dried, then examined by transmission electron microscopy at either 80 or 1000 kV. In contrast to air-dried material, critical point dried cells revealed each component clearly and with excellent contrast. All normal cytoplasmic structures (including coated vesicles, polyribosomes, microtubules and other fine components) were readily identifiable. Extensive structures such as microtubules and the endoplasmic reticulum (which appear fragmented in sections) were well displayed. At 1000 kV the beam readily penetrated even the thick nuclear and perinuclear cell regions and produced exceptionally crisp images. The methods described provide a simplified approach to the study of overall cell fine structure.

Cell Nucleolus↗