Activation of cyclic AMP-dependent kinase is required but may not be sufficient to mimic cyclic AMP-dependent DNA synthesis and thyroglobulin expression in dog thyroid cells.
Thyrotropin (TSH), via a cyclic AMP (cAMP)-dependent pathway, induces cytoplasmic retractions, proliferation, and differentiation expression in dog thyroid cells. The role of cAMP-dependent protein kinase (PKA) in the induction of these events was assessed by microinjection into living cells. Microinjection of the heat-stable inhibitor of PKA (PKI) inhibited the effects of TSH, demonstrating that activation of PKA was required in this process. Overexpression of the catalytic (C) subunit of PKA brought about by microinjection of the expression plasmid pC alpha ev or of purified C subunit itself was sufficient to mimic the cAMP-dependent cytoplasmic changes and thyroperoxidase mRNA expression but not to induce DNA synthesis and thyroglobulin (Tg) expression. The cAMP-dependent morphological effect was not observed when C subunit was coinjected with the regulatory subunit (RI or RII subunit) of PKA. To mimic the cAMP-induced PKA dissociation into free C and R subunits, the C subunit was coinjected with the regulation-deficient truncated RI subunit (RIdelta1-95) or with wild-type RI or native RII subunits, followed by incubation with TSH at a concentration too low to stimulate the cAMP-dependent events by itself. Although the cAMP-dependent morphology changes were still observed, neither DNA synthesis nor Tg expression was stimulated in these cells. Taken together, these data suggest that in addition to PKA activation, another cAMP-dependent mechanism could exist and play an important role in the transduction of the cAMP signal in thyroid cells.
Dremier
S
S
Institute of Interdisciplinary Research (I.R.I.B.H.N.), Université Libre de Bruxelles, Brussels, Belgium. Sdremier@ulb.ac.be
Pohl
V
V
Poteet-Smith
C
C
Roger
P P
PP
Corbin
J
J
Doskeland
S O
SO
Dumont
J E
JE
Maenhaut
C
C
D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000494:pharmacology*; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D004247:DNA / Q000096:biosynthesis; D004285:Dogs; D004789:Enzyme Activation; D005786:Gene Expression Regulation; D007453:Iodide Peroxidase / Q000096:biosynthesis; D008845:Microinjections; D010766:Phosphorylation; D011989:Receptors, Thyrotropin / Q000096:biosynthesis; D015398:Signal Transduction; D013954:Thyroglobulin / Q000096:biosynthesis*; D013961:Thyroid Gland / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D013972:Thyrotropin / Q000494:pharmacology*
D017362:Cyclic AMP Response Element-Binding Protein; D011989:Receptors, Thyrotropin; D013972:Thyrotropin; D004247:DNA; D013954:Thyroglobulin; D000242:Cyclic AMP; D007453:Iodide Peroxidase; D017868:Cyclic AMP-Dependent Protein Kinases