Normal hematopoiesis and inflammatory responses despite discrete signaling defects in Galpha15 knockout mice.
Galpha15 activates phospholipase Cbeta in response to the greatest variety of agonist-stimulated heptahelical receptors among the four Gq class G-protein alpha subunits expressed in mammals. Galpha15 is primarily expressed in hematopoietic cells in fetal and adult mice. We disrupted the Galpha15 gene by homologous recombination in embryonic stem cells to identify its biological functions. Surprisingly, hematopoiesis was normal in Galpha15(-/-) mice, Galpha15(-/-) Galphaq(-/-) double-knockout mice (which express only Galpha11 in most hematopoietic cells), and Galpha11(-/-) mice, suggesting functional redundancy in Gq class signaling. Inflammatory challenges, including thioglycolate-induced peritonitis and infection with Trichinella spiralis, stimulated similar responses in Galpha15(-/-) adults and wild-type siblings. Agonist-stimulated Ca(2+) release from intracellular stores was assayed to identify signaling defects in primary cultures of thioglycolate-elicited macrophages isolated from Galpha15(-/-) mice. C5a-stimulated phosphoinositide accumulation and Ca(2+) release was significantly reduced in Galpha15(-/-) macrophages. Ca(2+) signaling was abolished only in mutant cells pretreated with pertussis toxin, suggesting that the C5a receptor couples to both Galpha15 and Galphai in vivo. Signaling evoked by other receptors coupled by Gq class alpha subunits appeared normal in Galpha15(-/-) macrophages. Despite discrete signaling defects, compensation by coexpressed Gq and/or Gi class alpha subunits may suppress abnormalities in Galpha15-deficient mice.
Davignon
I
I
Pharmacology Department, UT Southwestern, Dallas TX 75235-9041, USA.
Catalina
M D
MD
Smith
D
D
Montgomery
J
J
Swantek
J
J
Croy
J
J
Siegelman
M
M
Wilkie
T M
TM
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D005333:Fetus; D043802:GTP-Binding Protein alpha Subunits, Gq-G11; D019204:GTP-Binding Proteins / Q000172:deficiency / Q000235:genetics / Q000502:physiology; D006410:Hematopoiesis / Q000235:genetics* / Q000502:physiology; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000502:physiology*; D020962:Heterotrimeric GTP-Binding Proteins / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D007249:Inflammation / Q000235:genetics* / Q000503:physiopathology; D017737:Macrophages, Peritoneal / Q000187:drug effects / Q000502:physiology*; D051379:Mice; D008807:Mice, Inbred BALB C; D018345:Mice, Knockout; D010538:Peritonitis / Q000503:physiopathology; D010716:Phosphatidylinositols / Q000378:metabolism; D011995:Recombination, Genetic; D015183:Restriction Mapping; D015398:Signal Transduction / Q000235:genetics; D013864:Thioglycolates / Q000633:toxicity; D017160:Trichinella spiralis; D014235:Trichinellosis / Q000503:physiopathology
D010716:Phosphatidylinositols; D013864:Thioglycolates; D019204:GTP-Binding Proteins; C112631:G protein alpha 16; D043802:GTP-Binding Protein alpha Subunits, Gq-G11; D020962:Heterotrimeric GTP-Binding Proteins; D002118:Calcium