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Vol. 11, Issue 3, 1077-1092, March 2000
Isoform of Protein Kinase C Is Involved in Signaling the
Response of Desmosomes to Wounding in Cultured Epithelial Cells
§ and
*School of Biological Sciences, University of Manchester,
Manchester, M13 9PT, United Kingdom; and Initiation of reepithelialization upon wounding is still poorly
understood. To enhance this understanding, we focus here on changes in
the adhesive state of desmosomes of cultured Madin-Darby canine kidney
cells in response to wounding of confluent cell sheets. Previous
results show that desmosomal adhesion in Madin-Darby canine kidney
cells changes from a calcium-dependent state to calcium independence in
confluent cell sheets. We show that this change, which requires culture
confluence to develop, is rapidly reversed upon wounding of confluent
cell sheets. Moreover, the change to calcium dependence in wound edge
cells is propagated to cells hundreds of micrometers away from the
wound edge. Rapid transition from calcium independence to calcium
dependence also occurs when cells are treated with phorbol esters that
activate PKC. PKC inhibitors, including the conventional isoform
inhibitor Gö6976, cause rapid transition from calcium dependence
to calcium independence, even in subconfluent cells. The cellular
location of the Division of
Cell Science, School of Biological Sciences, University of Southampton,
Southampton, SO16 7PX, United Kingdom
isoform of PKC correlates with the calcium
dependence of desmosomes. Upon monolayer wounding, PKC translocates
rapidly to the cell periphery, becomes Triton X-100 insoluble, and also becomes concentrated in lamellipodia. The PKC translocation upon wounding precedes both the increase in PKC activity in the membrane fraction and the reversion of desmosomes to calcium dependence. Specific depletion of PKC with an antisense oligonucleotide
increases the number of cells with calcium-independent desmosomes.
These results show that PKC participates in a novel signaling
pathway that modulates desmosomal adhesion in response to wounding.
Corresponding author. E-mail
address: david.garrod{at}man.ac.uk.
Present addresses: Research and Development,
Unipath, Priory Business Park, Bedford, MK44 3UP, UK;
§
Leukaemia Research Fund, Centre for Adult Leukaemia,
Department of Haematology, Imperial College School of Medicine,
Hammersmith Hospital, DuCane Road, London W12 ONN, UK.
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