Melatonin synthesis in the bovine pineal gland is regulated by type II cyclic AMP-dependent protein kinase.
We investigated the expression of regulatory (R) and catalytic (C) subunits of cyclic AMP-dependent protein kinase (cAK; ATP:protein phosphotransferase; EC 2.7.1.37) in the bovine pineal gland. In total RNA extracts of bovine pineal glands moderate levels of RI alpha/RII beta and high levels of C alpha and C beta mRNA were found. We were able to detect a strong signal for RII and C subunit at the protein level, whereas RI was apparently absent. Probing sections of the intact bovine pineal gland with RI and RII antibodies stained only RII in pinealocytes. Pairs of cyclic AMP analogues complementing each other in activation of type II cAK, but not cAKI-directed analogue pairs, showed synergistic stimulation of melatonin synthesis. Moreover, melatonin synthesis stimulated by the physiological activator norepinephrine in pineal cell cultures was inhibited by cAK antagonists. Taken together these results show the presence of RII regulatory and both C alpha and C beta catalytic subunits and thus cAKII holoenzyme in the bovine pineal gland. The almost complete inhibition of norepinephrine-mediated melatonin synthesis by the cAK antagonists emphasizes the dominant role of cyclic AMP as the second messenger and cAK as the transducer in bovine pineal signal transduction.
Maronde
E
E
Institute for Hormone and Fertility Research, University of Hamburg, Germany.
Middendorff
R
R
Telgmann
R
R
Müller
D
D
Hemmings
B
B
Taskén
K
K
Olcese
J
J
D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002417:Cattle; D002478:Cells, Cultured / Q000187:drug effects / Q000201:enzymology; D000242:Cyclic AMP / Q000819:agonists / Q000037:antagonists & inhibitors; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D007150:Immunohistochemistry; D007527:Isoenzymes / Q000378:metabolism*; D008550:Melatonin / Q000096:biosynthesis* / Q000235:genetics; D010870:Pineal Gland / Q000166:cytology / Q000201:enzymology*
D007527:Isoenzymes; D000242:Cyclic AMP; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D008550:Melatonin