Embryoid bodies from mouse stem cells express oxytocin receptor, Oct-4 and DAZL.
Oxytocin is a nine amino acid peptide involved in a wide spectrum of physiological functions; predominantly those concerning reproduction and differentiation are of interest. Oxytocin receptors are expressed at early developmental stages of mammals, suggesting that oxytocin might be involved in the determination of the germ stem cell line, at the very early stages of mammalian development. In this respect, the proximate aim of the present study was to confirm and further analyze the existence of oxytocin receptors at a very early level of cell commitment, that is, the determination of germ cells derived from embryoid bodies. To achieve our purpose we have cultured mouse embryonic stem cells under conditions inducing formation of embryoid bodies. In this work, ES cells were allowed to aggregate in a novel medium, "Stefanidis medium" from day 0 to day 14 until formed EBs. RNA was isolated from EBs and using RT-PCR we showed that EBs expressed Oct-4, OTR, OT, and DAZL. To demonstrate simultaneous expression immunocytochemistry was preformed, in which EBs showed strong immunoreactivity for both, OTR and DAZL molecular markers. We found that 35% of the cells displayed OTR, using flow cytometry. In addition, this novel medium showed to increase OTR mRNA. We propose, that at least in murine induced embryoid bodies there is simultaneous expression of oxytocin receptors and germ cell markers (DAZL) in many cells (expressing Oct-4). We thus conclude that, the oxytocin might indeed be a molecule playing a leading role in germ cell determination.
Stefanidis
Konstantinos
K
Laboratory of Stem Cells, Division of Reproductive Medicine, Alexandra Maternity Hospital, 1st Department of Obstetrics & Gynecology, Medical School, N&K University of Athens, 15 Neapoleos str., Marousi, Greece. kstefanidis@med.uoa.gr
Loutradis
Dimitris
D
Anastasiadou
Vasiliki
V
Beretsos
Panagiotis
P
Bletsa
Ritsa
R
Dinopoulou
Vasiliki
V
Lekka
Kaliopi
K
Elenis
Evangelia
E
Kiapekou
Erasmia
E
Koussoulakos
Stauros
S
Fotinos
Anastasios
A
Antsaklis
Aris
A
D000818:Animals; D002449:Cell Aggregation; D002454:Cell Differentiation; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D005434:Flow Cytometry; D018507:Gene Expression Regulation, Developmental; D007150:Immunohistochemistry; D051379:Mice; D050814:Octamer Transcription Factor-3 / Q000235:genetics* / Q000378:metabolism; D010121:Oxytocin / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D018045:Receptors, Oxytocin / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors
C508515:DAZL protein, mouse; D050814:Octamer Transcription Factor-3; D016601:RNA-Binding Proteins; D018045:Receptors, Oxytocin; D010121:Oxytocin