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Modulation of a volume-regulated chloride current by F-actin

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Abstract

We have examined whether F-actin integrity is involved in activation of a volume-regulated Cl− current (VRChlC) in B-lymphocytes. VRChlC activation was initiated in response to establishing a whole cell recording in the presence of a hyposmotic gradient. Parallel confocal microscopy experiments using Rhodamine Phalloidin (R-P) as a specific marker of F-actin showed that the submembrane actin ring is reversibly disrupted in response to an hyposmotic gradient. Disruptions of cortical F-actin integrity by 50 μm cytochalasin B (CB) does not trigger activation of VRChlC under isosmotic conditions or potentiate the rate of activation when the osmolarity of the extracellular solution was decreased by 75%. However, incubation with CB increased the rate of VRChlC activation in response to a 90% hyposmotic gradient. Phalloidin, a stabilizer of F-actin, decreases the rate of VRChlC activation in response to a 90% gradient, but has no effect in response to a 75% gradient. These observations suggest that disassembly of cortical F-actin is not critical for VRChlC activation in B-lymphocytes. The integrity of cortical F-actin, however, can exert a modulatory effect on the rate of VRChlC activation in the presence of a hyposmotic gradient.

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Authors and Affiliations

  1. Department of Physiology, Medical College of Pennsylvania and Hahnemann University, 19129, Philadelphia, PA

    I. Levitan, C. Almonte & S. S. Garber

  2. Department of Anatomy/Neurobiology, Medical College of Pennsylvania and Hahnemann University, 19129, Philadelphia, PA

    P. Mollard

Authors
  1. I. Levitan
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  2. C. Almonte
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  3. P. Mollard
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  4. S. S. Garber
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Additional information

We thank Mr. Gregory Kowalsky for excellent technical assistance and Drs. P. Hillman, B. Knight, M.P. Nusbaum and M. White for excellent suggestions and critical reading of the manuscript. Dr. Garber is an Established Investigator of the American Heart Association. This work was supported by NIDDK46672, AHA94002340, and NSF BIR-9413528.

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Levitan, I., Almonte, C., Mollard, P. et al. Modulation of a volume-regulated chloride current by F-actin. J. Membarin Biol. 147, 283–294 (1995). https://doi.org/10.1007/BF00234526

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