Heterogeneity of the changes in cytoplasmic pH upon serum stimulation of quiescent fibroblasts.

Journal of Cellular Physiology
G R BrightD L Taylor

Abstract

Addition of mitogens to quiescent cells results in rapid ionic changes in the cytoplasm, including pH. We studied the changes in cytoplasmic pH in single Swiss 3T3 cells upon serum stimulation using fluorescence ratio imaging microscopy. Quiescence was attained using two approaches, serum deprivation of subconfluent cells and confluence. All measurements were made in the presence of bicarbonate and the absence of other organic buffers. We also used BCECF coupled to dextran to avoid several artifacts associated with using BCECF-AM, including leakage and phototoxicity. Analysis of the changes in cytoplasmic pH demonstrated a dramatic heterogeneity in the responses of single cells. There were six basic classes of responses, 1) a fast alkalinization, reaching a maximum pH in approximately 2-5 min; 2) a slow alkalinization, reaching a maximum pH in 10-20 min; 3) a very slow alkalinization, not reaching a plateau pH within the measurement time; 4) no apparent change in pH during the measurement time; 5) an early transient acidification, followed by either a fast or slow alkalinization; and 6) an acidification, followed by alkalinization and then by a decrease to some intermediate pH. Subconfluent cells exhibited greater heterogeneity...Continue Reading

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Citations

Jan 2, 2007·Apoptosis : an International Journal on Programmed Cell Death·Carlos LizamaRicardo D Moreno
Feb 1, 1995·Current Opinion in Cell Biology·K A Giuliano, D L Taylor
Jul 1, 1992·Proceedings of the National Academy of Sciences of the United States of America·M A Schwartz, C Lechene
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