Background: During the progression of avian secondary palate morphogenesis, the rate of cell proliferation declines, whereas the production and accumulation of extracellular matrices increases. To investigate the regulation of these events, we examined the quail secondary palate for the activity of casein kinase 2 (CK 2), a pleiotropic serine/threonine second messenger independent enzyme implicated in cell growth and differentiation.
Methods: Quail palatal shelves were dissected between days 5 and 9 of incubation, which is the period of palate morphogenesis in quail, and prepared either for light microscopic observations or homogenized, cleared by ultracentrifugation, and then subjected to fractionation on a MonoQ column by fast protein liquid chromatography and Western immunoblotting.
Results: Histological examination showed that the palatal shelves appeared on day 5 of incubation and approximated by day 8 of incubation. Fractionation of palate extract using a Mono-Q column revealed the presence of a major peak of phosvitin phosphotransferase activity which eluted with 0.5 M NaCl. This activity peak coincided with the presence of a 42 kDa subunit of CK 2 as determined by Western blotting with a CK 2 specific antibody. The CK 2 activity towards phosvitin was elevated on days 5 and 6 and then rapidly declined by day 9. The decrease in CK 2 activity did not correlate with a decrease in CK 2 protein during palate development indicating that the differential activity of the CK 2 enzyme observed during quail palate development may be due to post-translational modifications of the enzyme. A high positive correlation was found between the CK 2 phosphotransferase activity and both the proliferation index and DNA synthesis during palate development.
Conclusion:
On the basis of literature analysis and the results of the present study, it was suggested that the activity of CK 2 may be regulated along with protein kinase A to coordinate cell proliferation and the synthesis of extracellular matrices during palate development in quail.