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Journal of Cell Science, Vol 87, Issue 3 389-398, Copyright © 1987 by Company of Biologists
JOURNAL ARTICLES |
EJ Sanders and P Chokka
Department of Physiology, University of Alberta, Edmonton, Canada.
Chick embryos in culture were treated with the secretion-inhibiting ionophore monensin at the gastrulation stage of development. After treatment for 5 h at a concentration of 10(-5) M the embryos showed a drastic reduction in the tissue space between the upper and lower epithelia, and reduced spreading of the mesoderm cells that occupied this space. The basement membrane, to which many mesoderm cells are normally attached, showed varying degrees of disruption, which permitted blebbing of the overlying epithelium. Intracellularly, the treatment caused extensive vacuolization in all tissues. These results were consistent with the effects expected by removal of hyaluronic acid from the extracellular space, and this was confirmed by demonstrating a sharp reduction in glucosamine incorporation in this region. Examination of the effects of monensin on isolated mesoderm cells in culture using interference reflection microscopy indicated that the spreading of these cells was reduced independently of the changes in the extracellular matrix of the embryo. That this was probably due to the inhibition of cell surface fibronectin secretion was shown by demonstrating severe changes in the distribution of this glycoprotein using the immunofluorescent technique. It is concluded that the effects of monensin on this intact, developing system are due primarily to the disruption of hyaluronic acid secretion, but that disrupted fibronectin synthesis contributes to the reduced spreading of mesoderm cells.