Epigenetic deregulation through DNA demethylation seems not to interfere with the differentiation of epithelia from pre-gastrulating rat embryos in vitro.
One of the epigenetic mechanisms controlling differentiation during mammalian development is the process of DNA methylation. The differentiation of tissues in pre-gastrulating rat embryos cultivated in vitro under the influence of the demethylating agent 5-azacytidine (5azaC) was investigated. Eight-day-old Fisher rat embryos consisting of epiblast and hypoblast (primitive ectoderm and primitive endoderm) were isolated and cultivated in serum-supplemented medium by air-lifting method in vitro. A single dose of 5azaC (30 microM) was added to the culture medium on day 5 of cultivation. After 14 days, teratoma-like structures developed and were processed by routine histology. When compared to controls, the explants treated with 5azaC showed a statistically significant higher incidence of neuroblasts, myotubes, cartilage, and blood islands. On the other hand, the incidence of stratified squamous, columnar and glandular epithelium was not statistically different from controls. It seems that differentiation of epithelia was not sensitive to DNA demethylation caused by 5azaC like differentiation of other tissues, especially mesodermal derivatives.
Vlahović
Maja
M
Department of Biology School of Medicine, University of Zagreb, Salata 3 HR-10000 Zagreb, Croatia. majav@mef.hr
Bulić-Jakus
Floriana
F
Jurić-Lekić
Gordana
G
Serman
Ljiljana
L
Sincić
Nino
N
Katusić
Ana
A
D000818:Animals; D001374:Azacitidine / Q000494:pharmacology*; D019175:DNA Methylation / Q000187:drug effects*; D004622:Embryo, Mammalian; D044127:Epigenesis, Genetic / Q000187:drug effects*; D004848:Epithelium / Q000187:drug effects / Q000196:embryology*; D054262:Gastrulation; D051381:Rats; D011916:Rats, Inbred F344; D046509:Tissue Culture Techniques
D001374:Azacitidine