The morphology of erythroid cells separated by density gradient centrifugation through ficoll |
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Authors: | Dr. K. Brasch G. H. M. Adams J. M. Neelin |
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Affiliation: | (1) Departments of Biology, Queen's University, Kingston and Carleton University, Ottawa, Ontario, Canada;(2) Department of Biology, Queen's University, K7L 3N6 Kingston, Ontario, Canada |
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Abstract: | Summary The regenerating blood of geese injected with phenylhydrazine was subjected to large scale, zonal centrifugation through density gradients of Ficoll. In this way, erythroid cells were fractionated according to their respective stages of development. Highly enriched fractions were obtained, containing cells that were well preserved as assessed by both light and electron microscopy. The separated cells exhibited ribosome density and nucleic acid and protein staining patterns typically associated with erythrocyte differentiation. Morphometric analysis of nuclei indicated that despite an apparent net increase in the amount of compact chromatin during development, comparatively little difference existed between the volumes of condensed chromatin present in immature and mature cells. Instead, there was a three fold decrease in nuclear volume between young erythroblasts and reticulocytes, coupled with a concomitant decrease in the volume occupied by dispersed chromatin, RNP and nucleoli. These observations are discussed in relation to molecular changes associated with nuclear differentiation in erythroid cells.Supported by grants from the National Research Council of CanadaWe thank Dr. G. Setterfield for assistance with the EM data and we are grateful to the N.R.C. for use of centrifuges and the zonal rotor |
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Keywords: | Erythrocytes Zonal centrifugation Microscopy, chromatin content Morphometry |
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