Abstract
P19 mouse embryonal carcinoma cells can be stimulated to differentiate into endodermal-like, mesodermal-like, and neuronal-like phenotypes in response to specific morphogens. At low concentrations, retinoic acid stimulates P19 embryonal cells to differentiate to cells displaying an endodermal phenotype, whereas at higher concentrations it stimulates differentiation to neuroectoderm. The Galpha12 and Galpha13 subunits of heterotrimeric G-proteins are expressed in the embryonal P19 cells and stimulated in response to retinoic acid as the cells differentiate to endodermal or neuroectodermal phenotypes. Suppression of the expression of either Galpha12 or Galpha13 by antisense RNA is shown to promote cell detachment from substratum and apoptosis. Expression of the constitutively active, mutant form of Galpha12 (Q229L), in contrast, stimulates loss of the embryonal phenotype. Expression of the constitutively active form of Galpha13 (Q226L) stimulates differentiation of the cells from embryonal to endodermal, in the absence of retinoic acid. Thus, both Galpha12 and Galpha13 are essential to stimulation of cell differentiation by retinoic acid. Deficiency of either Galpha12 or Galpha13 increases programmed cell death.
Highlights
Heterotrimeric G-proteins1 play important roles in cell signaling and more complex biological responses such as oncogenesis, cell differentiation, and development [1,2,3,4]
P19 mouse embryonal carcinoma cells are sensitive to aggregation and retinoic acid (RA), differentiating to an endoderm phenotype in response to low concentrations of RA (10 nM) and to a neuroectodermal-like phenotype in response to high concentration of RA (100 nM)
Aggregation on bacterial-grade Petri dishes for 4 days followed by dissociation and 3 days further growth on tissue culture plates is required for stimulation of the differentiated states by RA [18]
Summary
Heterotrimeric G-proteins1 play important roles in cell signaling and more complex biological responses such as oncogenesis, cell differentiation, and development [1,2,3,4]. The expression of G␣12 and G␣13 mRNAs in P19 cells was investigated in cells allowed to aggregate for 4 days in either the absence or the presence of increasing concentrations of RA and dissociated and grown further for 3 days on tissue culture plates (Fig. 2).
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
