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PubMed1982

Golgi detection in mitotic and interphase cells by antibodies to secreted galactosyltransferase.

Secreted galactosyltransferase from bovine milk was used to induce antibodies cross-reacting with corresponding intracellular enzymes in a variety of cell lines and tissues. In contrast to the original antigen, the reactive intracellular galactosyltransferase appears as individual species (apparent MW approx. 42000-46000) in SDS-polyacrylamide gel electrophoresis. In indirect immunofluorescence microscopy affinity-purified IgGs locate the galactosyltransferase in a distinct perinuclear and juxtanuclear position indicative for the Golgi region. The rearrangement of labelled structures upon colcemid or monensin treatment--drugs known to influence Golgi morphology and function--is further proof for a Golgi association. The fate and distribution of Golgi elements during mitosis is described at the light microscopical level using galactosyltransferase as easily identifiable marker. In addition we evaluate the utilization of wheat germ agglutinin (WGA) binding for Golgi identification on tissue culture cells and show that WGA is not a reliable marker for certain cell types such as MDCK.

Animals↗
PubMed1989

Induced interphase cell retraction: its reversal and EGF potentiation.

Human epidermoid A431 carcinoma cells retract into globular shapes upon activation with epidermal growth factor (EGF) but not normal cells. We find that normal anchor-dependent cultured cells will retract from attachment on incubation in saline with pH upshifting. There is significant potentiation of that phenomenon by EGF prepulsing as seen in (a) dose- and time-response experiments quantitated by measuring profile area and perimeter of cells (b) 22Na+-uptake studies and (c) pHi measurements. EGF action by itself produces quantitatively significant retractions, although only to a very small degree. Blocking Na+/H+ exchanges using (a) trifluoperazine (150 microM), (b) amiloride (2 mM) and (c) Na+-omission could block the cell retractions. Retracted cells can be reversed in low [Na+]o during which rapid 22Na+-efflux is observed. A dose-response ouabain (1 mM) sensitive relationship exists between [Na+]o and retardation of the reversal phenomenon. Ouabain however does not prevent reversal in low [Na+]o, nor even in combination with high [Na+]o given the presence of Ca2+. Retraction into spherical shapes would maximize volume-holding for a given surface area, however they are quantitatively more bulgy than cells rounded by trypsinization.

Amiloride↗
PubMed1993

Interphase genome as the active space: chromatin dynamics during chick embryo chondrogenesis.

The rearrangement of the chromatin that takes place during cytodifferentiation was studied using TV image analysis in chick limb bud cartilage stained for DNA. The redistribution of the chromatin was compatible with the Rabl orientation: chromatin was extended radially from the centromeric ring to the telomere pole in young chondroblasts, and contracted back in ageing chondrocytes. The direction and gradient of this redistribution correlate with the changes in DNA content within the chromocentres formed by pericentromeric heterochromatin. In turn, intercalary heterochromatin regulates the condensation of the adjacent euchromatin depending upon the position in this radial-polar gradient.

Animals↗