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Involvement of mitogen-activated protein kinase and casein kinase 2 in regulation of cell proliferation during development of the secondary palate in quail

University of British Columbia

A study was undertaken to investigate: (1) the kinetics of cell proliferation; (2) the activity of two protein kinases, MAPK and CK2, that are implicated in the regulation of cell proliferation; (3) the effect of a growth factor, EGF, on the proliferation of cultured quail palate mesenchymal cells (QPMC); and (4) the activity of MAPK and CK2 following E G F treatment of cultured QPMC. Initially, an in vivo/in vitro comparison of developing quail was performed to validate the whole embryo culture technique. The data showed that, on the basis of C R L and developmental staging method, the in vitro (whole embryo culture) embryos developed in a manner similar to their in vivo counterparts. The whole embryo culture was then used to analyze the cell proliferation kinetics by 3H-thymidine autoradiography during quail palate morphogenesis. The results indicated that the rate of cell proliferation declined with the advancing morphogenesis of quail palate. Subsequently, embryonic palates were dissected at 24 hour intervals between days 5 and 9 of incubation and homogenized. Isolated proteins from the homogenate were fractionated by FPLC, and analyzed by phosphotransferase assays, scintillation counting, and Western blotting to detect the presence and activation of two second-messenger independent protein kinases, MAPK and CK2, implicated in the regulation of cell proliferation. The results showed that phosphotransferase activity towards MBP showed a negative, whereas that towards phosvitin showed a positive correlation with proliferation of QPMC. Western blotting indicated that both MAPK and C K 2 were present during quail palate development. Finally, growth behaviour of QPMC in primary cell culture was analyzed. The results showed that the number of Q P M C increased faster in EGF-treated than in control cultures. E G F stimulated phosphotransferase activity towards MBP and phosvitin, and Western blotting indicated that both MAPK and CK2 were activated. It was suggested that both MAPK and CK2 may be involved in growth factor regulated cell proliferation during the palate development.

Thesis/Dissertation

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2009-03-11

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http://hdl.handle.net/2429/5876

Oral Biology and Medical Sciences

University of British Columbia

UBCV

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