Expression and activity of protein kinase C isoenzymes during normal and abnormal murine palate development.
Protein kinase C (PKC) plays a critical role in signal transduction, mediating various cellular events critical for normal development, including that of the palate. In vivo and in vitro studies suggest the relevance of the inhibition of PKC by the mycotoxin, secalonic acid D (SAD), to its induction of cleft palate (CP) in mice. In the present study, temporal and spatial expression and the activity of various PKC isoenzymes were studied in the control and SAD-exposed murine embryonic palate during gestational days (GD) 12-14.5 by western blotting, immunohistochemistry, and phosphotransfer assay. The Ca2+-dependent isoenzymes, PKC alpha and PKC betaII, showed significant expression on GD 12.0, which gradually decreased through GD 14.5, whereas PKC betaI and PKC gamma were negligible throughout. All Ca2+-independent isoenzymes (epsilon, delta, and zeta) were expressed more abundantly and, in contrast to the Ca2+-dependent ones, progressively increased with age. SAD failed to alter this pattern of expression but enhanced the phosphorylation of PKC epsilon throughout development. Immunohistochemical analysis revealed an isoenzyme-specific distribution of PKC between the epithelium and mesenchyme. As expected, SAD significantly inhibited the total Ca2+-dependent PKC activity in palatal extracts. Although total Ca2+-independent PKC activity in palatal extracts was unaffected by SAD, individual pure isoenzymes were either selectively inhibited (PKC zeta), stimulated (PKC delta), or unaffected (PKC epsilon) by SAD. These results show that PKC isoenzymes exhibit dynamic temporal and spatial patterns of expression and activity in the developing palate and that the induction of CP by SAD is associated with an alteration in their activation and/or activity.
Balasubramanian
G
G
Department of Veterinary Biomedical Sciences, University of Missouri-Columbia, 65211, USA.
Amann
J F
JF
Reddy
C S
CS
D000818:Animals; D015153:Blotting, Western; D002972:Cleft Palate / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D004305:Dose-Response Relationship, Drug; D005260:Female; D007150:Immunohistochemistry; D007527:Isoenzymes / Q000378:metabolism; D008297:Male; D051379:Mice; D010159:Palate / Q000002:abnormalities / Q000196:embryology / Q000201:enzymology; D010766:Phosphorylation; D010770:Phosphotransferases / Q000378:metabolism; D011493:Protein Kinase C / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism*; D064546:Protein Kinase C beta; D051571:Protein Kinase C-alpha; D051745:Protein Kinase C-delta; D051744:Protein Kinase C-epsilon; D015398:Signal Transduction; D013997:Time Factors; D014018:Tissue Distribution; D014966:Xanthenes / Q000378:metabolism / Q000494:pharmacology; D044004:Xanthones
D007527:Isoenzymes; D014966:Xanthenes; D044004:Xanthones; C009218:secalonic acid; D010770:Phosphotransferases; C497259:Prkcd protein, mouse; C497256:Prkce protein, mouse; C496630:Prkca protein, mouse; D011493:Protein Kinase C; D064546:Protein Kinase C beta; D051571:Protein Kinase C-alpha; D051745:Protein Kinase C-delta; D051744:Protein Kinase C-epsilon