Disruption of the mouse mTOR gene leads to early postimplantation lethality and prohibits embryonic stem cell development.
The mammalian target of rapamycin (mTOR) is a key component of a signaling pathway which integrates inputs from nutrients and growth factors to regulate cell growth. Recent studies demonstrated that mice harboring an ethylnitrosourea-induced mutation in the gene encoding mTOR die at embryonic day 12.5 (E12.5). However, others have shown that the treatment of E4.5 blastocysts with rapamycin blocks trophoblast outgrowth, suggesting that the absence of mTOR should lead to embryonic lethality at an earlier stage. To resolve this discrepancy, we set out to disrupt the mTOR gene and analyze the outcome in both heterozygous and homozygous settings. Heterozygous mTOR (mTOR(+/-)) mice do not display any overt phenotype, although mouse embryonic fibroblasts derived from these mice show a 50% reduction in mTOR protein levels and phosphorylation of S6 kinase 1 T389, a site whose phosphorylation is directly mediated by mTOR. However, S6 phosphorylation, raptor levels, cell size, and cell cycle transit times are not diminished in these cells. In contrast to the situation in mTOR(+/-) mice, embryonic development of homozygous mTOR(-/-) mice appears to be arrested at E5.5; such embryos are severely runted and display an aberrant developmental phenotype. The ability of these embryos to implant corresponds to a limited level of trophoblast outgrowth in vitro, reflecting a maternal mRNA contribution, which has been shown to persist during preimplantation development. Moreover, mTOR(-/-) embryos display a lesion in inner cell mass proliferation, consistent with the inability to establish embryonic stem cells from mTOR(-/-) embryos.
Genome Research Institute, University of Cincinnati, 2180 E. Galbraith Rd., Cincinnati, OH 45237, USA.
Yann-Gaël
YG
Gangloff
Matthias
M
Mueller
Stephen G
SG
Dann
Petr
P
Svoboda
Melanie
M
Sticker
Jean-Francois
JF
Spetz
Sung Hee
SH
Um
Eric J
EJ
Brown
Silvia
S
Cereghini
George
G
Thomas
Sara C
SC
Kozma
D000483:Alleles; D000818:Animals; D002454:Cell Differentiation; D002455:Cell Division; D048429:Cell Size; D002478:Cells, Cultured; D020964:Embryo Loss / Q000235:genetics* / Q000473:pathology*; D047108:Embryonic Development / Q000502:physiology*; D005260:Female; D005347:Fibroblasts; D017353:Gene Deletion; D006579:Heterozygote; D051379:Mice; D018345:Mice, Knockout; D011247:Pregnancy; D011494:Protein Kinases / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism*; D013234:Stem Cells / Q000378:metabolism* / Q000473:pathology*; D058570:TOR Serine-Threonine Kinases
D011494:Protein Kinases; C546843:mTOR protein, mouse; D058570:TOR Serine-Threonine Kinases