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PKCδ activation promotes endothelial substrate adhesion.
Author(s) -
Alexander Jonathan Steven,
Carpenter April C
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.5.a1168
Subject(s) - protein kinase c , adhesion , bryostatin 1 , substrate (aquarium) , microbiology and biotechnology , chemistry , biophysics , biochemistry , signal transduction , biology , ecology , organic chemistry
We previously showed that PMA and bryostatin‐1 regulate neutrophil (PMN) transendothelial migration (TEM) through activation of PKCä. We also reported that endothelial barrier (thought to regulate TEM) and substrate adhesion are reciprocally related. We previously studied effects of these drugs on barrier; here we examined substrate adhesion. Our prior data show that PMA decreased TEER, while bryostatin‐1 did not. Treatment with 100nM PMA or Bryostatin‐1 (1 hr) increased substrate adhesion, as measured by resistance to trypsin, from 54.71% to 81.89% and 77.58%, respectively. Since bryostatin‐1 preferentially activates the ä, å isoforms, and we have shown no involvement for the å isoform in PMN TEM, we studied PKCä in bryostatin‐induced substrate adhesion. Endothelium were pretreated with PKC inhibitors Go‐6976 (á, â), and Go‐6983 (á, â, ä) at 10nM and 100nM 1 hr prior to 100nM bryostatin‐1. 100nM Go‐6976 reduced substrate adhesion by 9.74 % (p<0.05), but 10nM did not reduce substrate adhesion. However, both 10nM and 100nM Go‐6983 reduced bryostatin‐1‐induced substrate adhesion (22.21% and 23.14%, respectively, p<0.001). Using PKCä siRNA nucleofection, we saw that PKCä siRNA reduced bryostatin‐1‐induced substrate adhesion by 26.67% (p<0.001). Although several isoforms of PKC may contribute to bryostatin‐1 induced substrate adhesion, our data suggest a central role for the ä isoform. Thus, endothelial substrate adhesion may be altered independent of barrier, and is a possible factor regulating PMN TEM. This study supported by NIH grant DK43785.