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Constitutively active mutant Gsα (G22T) and null-mutant Gαi-2 (G203T) induce primitive endoderm from stem cells

Gao, Ping, David C. Watkins, and Craig C. Malbon. Constitutively active mutant G s α (G225T) and null-mutant Gα i-2 (G203T) induce primitive endoderm from stem cells. Am. J. Physiol. 268 (Cell Physiol. 37) : C1460-C1466, 1995.-In F9 teratocarcinoma stem cells, retinoic acid induces a primitive endoderm-like phenotype and a sharp decline in Gα i-2 , a response mimicked by expression of RNA antisense to Gα i-2 in the absence of this morphogen (D. C. Watkins, G. L. Johnson, and C. C. Malbon. Science Wash. DC 258 : 1373-1375, 1992). The role of the G s α/Gα i-2 axis in cellular differentiation was explored. In the absence of retinoic acid, F9 stem cells stably expressing a constitutively active mutant of G s α (G225T) progressed to the primitive endoderm phenotype, as judged by morphological and differentiation markers, such as tissue plasminogen activator. Although elevated in cells expressing G225T Gas, adenosine 3',5'-cyclic monophosphate does not mimic retinoic acid action and alone fails to induce stem cells to primitive endoderm. In the absence of retinoic acid, expression of a null mutant of Gα i-2 (G203T) also induced stem cells to primitive endoderm. These observations establish G proteins in the G s α/Gα i-2 axis as a control point for regulating progression to primitive endoderm independent of adenylate cyclase, in the present study's model of early mouse development.