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M202130200v1
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Papers In Press, published online ahead of print October 9, 2002
J. Biol. Chem, 10.1074/jbc.M202130200
Submitted on March 4, 2002
Revised on October 3, 2002
Accepted on October 9, 2002

Growth hormone-induced diacylglycerol and ceramide formation via Galpha i3 and Gbeta gamma in GH4 pituitary cells: Potentiation by dopamine-D2 receptor activation

Gele Liu, Liliane Robillard, Behzad Banihashemi, and Paul R. Albert

Department of Neuroscience, Ottawa Health Research Institute, Ottawa, Ontario K1H-8M5

Corresponding Author: palbert{at}uottawa.ca

Growth hormone (GH) secretion is regulated by indirect negative feedback mechanisms. To address whether GH has direct actions on pituitary cells, lipid signaling in GH4ZR7 somatomammotroph cells was examined. GH (EC50 = 5 nM) stimulated diacylglycerol (DAG) and ceramide formation in parallel by over 10-fold within 15 min and persisting for >3 h. GH-induced DAG/ceramide formation was blocked by pertussis toxin (PTX) implicating Gi/Go proteins and was potentiated 1.5-fold by activation of Gi/Go-coupled dopamine-D2S receptors, which had no effect alone. Following PTX pretreatment, only PTX-resistant Galpha i3, not Galpha o or Galpha i2, rescued GH-induced DAG/ceramide signaling. GH-induced DAG/ceramide formation was also blocked in cells expressing Gbeta gamma blocker GRK-ct. In GH4ZR7 cells, GH induced phosphorylation of JAK2 and STAT5, which was blocked by PTX and mimicked by ceramide analogue C2-ceramide or sphingomyelinase treatment to increase endogenous ceramide. We conclude that in GH4 pituitary cells, GH induces formation of DAG/ceramide via a novel Galpha i3/Gbeta gamma -dependent pathway. This novel pathway suggests a mechanism for autocrine feedback regulation by GH of pituitary function.


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