Arachidonic acid release by ionomycin and phorbol ester is similar in C127 epithelial cells expressing wild-type or mutated (delta F508) cystic fibrosis transmembrane conductance regulator.
The Ca2+ ionophore ionomycin induced cytosolic [Ca2+ ]i elevation as well as strong activation of Cl- efflux in mouse mammary epithelial cell lines expressing wild-type or mutated (deletion of phenylalaline 508) cystic fibrosis transmembrane conductance regulator (CFTR) or vector. Ionomycin-induced Cl- efflux was abolished by the intracellular Ca2+ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid, whereas both activators and inhibitors of phospholipase A2 had no effect, indicating the involvement of Ca2+-dependent Cl- channels. Stimulation of arachidonic acid release by ionomycin and phorbol ester was not significantly different between wild-type or mutated cell lines, whereas vector-transfected cells exhibited a significant higher release, which was shown to be due to larger amount of immunoreactive cytosolic phospholipase A2. These results indicate that phospholipase A2 activity of C127 cells was not influenced by the presence of wild-type or mutated CFTR.
Porcelli
A M
AM
Dip di Biologia ES, Università di Bologna, Italy.
Ghelli
A
A
Mastrocola
T
T
Rugolo
M
M
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D000818:Animals; D016718:Arachidonic Acid / Q000096:biosynthesis*; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D004847:Epithelial Cells / Q000378:metabolism*; D015151:Immunoblotting; D015759:Ionomycin / Q000494:pharmacology*; D007476:Ionophores / Q000494:pharmacology; D007477:Ions; D051379:Mice; D009154:Mutation; D010703:Phorbol Esters / Q000494:pharmacology*; D010741:Phospholipases A / Q000378:metabolism; D054467:Phospholipases A2; D013050:Spectrometry, Fluorescence; D013997:Time Factors; D014162:Transfection
D007476:Ionophores; D007477:Ions; D010703:Phorbol Esters; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D005576:Colforsin; D016718:Arachidonic Acid; D015759:Ionomycin; D000242:Cyclic AMP; D010741:Phospholipases A; D054467:Phospholipases A2; D002118:Calcium; D015056:1-Methyl-3-isobutylxanthine