It is known that several hormonal primary messengers are capable of regulating developmentally significant biological responses in embryonic palatal tissue by modulating intracellular levels of cAMP. The effects of cAMP on metabolic pathways in eukaryotes are mediated by cAMP- dependent protein kinases (cAMP-dPK) and we have demonstrated spatial and temporal alterations in these enzymes in tissue of the developing mammalian secondary palate. Our studies characterizing cAMP-dPK in tissue derived from the developing mammalian palate has allowed consideration of cAMP-dPK as a key regulatory enzyme, capable of transducing hormonally elevated intracellular levels of cAMP into metabolic responses during orofacial ontogenesis. Our overall objectives in this application include: 1) Determination of whether the temporal alterations in soluble cAMP-dPK that we have defined are the result of transcriptional regulation of cAMP-dPK gene expression; 2) Utilizing a variety of """"""""blocking"""""""" strategies (introduction of A) antisense RNA or b) a recombinant expression vector encoding cAMP-dPK inhibitor protein or c) a membrane permeable specific competetive inhibitor of cAMP-dPK types I and II) to precisely define a functional role for cAMP-dPK in palate medial edge epithelial cell differentiation, mesenchymal cell proliferation and/or GAG synthesis; 3) Analysis of the regulation of cAMP-dPK regulatory subunit gene expression by transfection of cAMP-dPK regulatory and catalytic subunits into palate mesenchymal cells; 4) Determination of the presence and distribution of a cyclic AMP response element-binding protein (CREBP) in developing embryonic palatal tissue. These studies will directly test hypotheses regarding the functional role that cAMP-dPK plays in various aspects of palatal tissue differentiation and define the means by which cellular regulatory signals affect gene expression during embryonic palate development.

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Project (R01)
Project #
5R01DE008199-06
Application #
3222018
Study Section
Oral Biology and Medicine Subcommittee 1 (OBM)
Project Start
1988-07-01
Project End
1995-06-30
Budget Start
1993-07-01
Budget End
1994-06-30
Support Year
6
Fiscal Year
1993
Total Cost
Indirect Cost
Name
Thomas Jefferson University
Department
Type
Schools of Medicine
DUNS #
061197161
City
Philadelphia
State
PA
Country
United States
Zip Code
19107
Nugent, P; Greene, R M (1998) Use of antisense oligonucleotides to study the role of CRABPs in retinoic acid-induced gene expression. Methods Mol Biol 89:191-202
Weston, W M; Greene, R M (1995) Developmental changes in phosphorylation of the transcription factor CREB in the embryonic murine palate. J Cell Physiol 164:277-85
Nugent, P; Potchinsky, M; Lafferty, C et al. (1995) TGF-beta modulates the expression of retinoic acid-induced RAR-beta in primary cultures of embryonic palate cells. Exp Cell Res 220:495-500
Greene, R M; Lloyd, M R; Uberti, M et al. (1995) Patterns of cyclic AMP-dependent protein kinase gene expression during ontogeny of the murine palate. J Cell Physiol 163:431-40
Nugent, P; Greene, R M (1994) Interactions between the transforming growth factor beta (TGF beta) and retinoic acid signal transduction pathways in murine embryonic palatal cells. Differentiation 58:149-55
Gehris, A L; Pisano, M M; Nugent, P et al. (1994) Regulation of TGF beta 3 gene expression in embryonic palatal tissue. In Vitro Cell Dev Biol Anim 30A:671-9
Gawel-Thompson, K J; Greene, R M (1992) Quantification and localization of ornithine decarboxylase in the embryonic palate. J Exp Zool 261:441-50
Greene, R M; Lloyd, M R; Pisano, M M (1992) Cyclic AMP-dependent protein kinase in human embryonic palate mesenchymal cells. In Vitro Cell Dev Biol 28A:755-62
Gehris, A L; Greene, R M (1992) Regulation of murine embryonic epithelial cell differentiation by transforming growth factors beta. Differentiation 49:167-73
Weston, W M; Greene, R M (1991) Effects of ethanol on cAMP production in murine embryonic palate mesenchymal cells. Life Sci 49:489-94

Showing the most recent 10 out of 21 publications