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Calmodulin and wound healing in the coenocytic green alga Ernodesmis verticillata (Kützing) Børgesen

Immunofluorescence and effects of antagonists

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Abstract

The involvement of calmodulin (CaM) in wound-induced cytoplasmic contractions in E. verticillata was investigated. Indirect immunofluorescence of CaM in intact cells showed a faint, reticulate pattern of fluorescence in the cortical cytoplasm. Diffuse fluorescence was evident deeper within the cytoplasm. In contracted cells, CaM co-localizes with actin in the cortical cytoplasm in extensive, longitudinal bundles of microfilaments (MFs), and in an actin-containing reticulum. No association of CaM with tubulin was ever observed in the cortical cytoplasm at any stage of wound-healing. When contraction rates in wounded cells are measured, a lag period of 2 min is followed by a rapid, steady rate of movement over the subsequent 10 min. The delay in the initiation of longitudinal contraction corresponds to the time necessary for the assembly of the longitudinal MF bundles. Cytoplasmic motility was inhibited in a dose-dependent manner by CaM antagonists. In these inhibited cells, MF bundles did not assemble, or were poorly formed. In the latter case, CaM was always found associated with MFs. These results indicate a direct spatial and temporal correlation between CaM and actin, and a potential role for CaM in regulating the formation of functional MF bundles during wound-induced cytoplasmic contraction in Ernodesmis.

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Abbreviations

CaM:

calmodulin

DMSO:

dimethyl sulfoxide

EGTA:

ethylene glycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid

MF(s):

microfilament(s)

MT(s):

microtubule(s)

TFP:

trifluoperazine

w-5:

N-(6-aminohexyl)-1-naphthalenesulfonamide

W-7:

N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide

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Author notes

  1. R. H. Goddard

    Present address: Department of Plant Biology, 220 Biological Sciences Center, University of Minnesota, 1445 Gortner Ave., 55108, Paul, MN, USA

Authors and Affiliations

  1. Department of Botany, University of Texas, 78713, Austin, TX, USA

    R. H. Goddard & J. W. La Claire II

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Additional information

We are especially grateful to: Dr. J.A. West (University of California, Berkeley) for the original algal isolates; Dr. L. Van Eldik (Vanderbilt University School of Medicine) and Dr. J.L. Lessard (University of Cincinnati College of Medicine) for graciously providing CaM and actin antibodies, respectively; Dr. S.J. Roux (University of Texas, Austin) for the gift of purified oat CaM; Dr.H. Green (Smith, Kline and French Laboratories, Philadelphia, Penn., USA) for providing the trifluoperazine; and M.E.T. Scioli for assistance with the statistical analyses. Portions of this work were supported by National Science Foundation grant DCB 8402345 and U.S. Department of Agriculture grant 87-CRCR-1-2545 to J.W.L.

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Goddard, R.H., La Claire, J.W. Calmodulin and wound healing in the coenocytic green alga Ernodesmis verticillata (Kützing) Børgesen. Planta 183, 281–293 (1991). https://doi.org/10.1007/BF00197800

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