Embryoid bodies from mouse stem cells express oxytocin receptor, Oct-4 and DAZL
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. Bio Systems
(Impact Factor: 1.55).
09/2009; 98(2):122-6. DOI: 10.1016/j.biosystems.2009.08.004
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.
Available from: PubMed Central
- "OXTR is expressed at early developmental stages of mammals, such as in human amniotic fluid cells  and cultured mouse embryonic stem cells . Thus, OXT could participate in the differentiation of the germ stem cell line at the very early stages of mammalian development . "
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ABSTRACT: Oxytocin (OXT) is a hypothalamic neuropeptide composed of nine amino acids. The functions of OXT cover a variety of social and nonsocial activity/behaviors. Therapeutic effects of OXT on aberrant social behaviors are attracting more attention, such as social memory, attachment, sexual behavior, maternal behavior, aggression, pair bonding, and trust. The nonsocial behaviors/functions of brain OXT have also received renewed attention, which covers brain development, reproduction, sex, endocrine, immune regulation, learning and memory, pain perception, energy balance, and almost all the functions of peripheral organ systems. Coordinating with brain OXT, locally produced OXT also involves the central and peripheral actions of OXT. Disorders in OXT secretion and functions can cause a series of aberrant social behaviors, such as depression, autism, and schizophrenia as well as disturbance of nonsocial behaviors/functions, such as anorexia, obesity, lactation failure, osteoporosis, diabetes, and carcinogenesis. As more and more OXT functions are identified, it is essential to provide a general view of OXT functions in order to explore the therapeutic potentials of OXT. In this review, we will focus on roles of hypothalamic OXT on central and peripheral nonsocial functions.
07/2013; 2013(4429). DOI:10.1155/2013/179272
Available from: link.springer.com
- "Therefore, the hypothesis that the stem cells found in granulosa cells cannot be differentiated in germ cells is strongly supported because of the lack of DAZL, nanog, vasa and stella gene expression. In addition, Stefanidis et al. (2009) found that at least in murine induced embryoid bodies there is simultaneous expression of oxytocin receptors and germ cell markers (DAZL) in many cells "
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The purpose of the study was to determine the incidence of gene expression of Oct-4 and DAZL, which are typical markers for stem cells, in human granulosa cells during ovarian stimulation in women with normal FSH levels undergoing IVF or ICSI and to discover any clinical significance of such expression in ART.
Twenty one women underwent ovulation induction for IVF or ICSI and ET with standard GnRH analogue-recombinant FSH protocol. Infertility causes were male and tubal factor. Cumulus–mature oocyte complexes were denuded separately and granulosa cells were analyzed for each patient separately using quantitative reverse-transcription–polymerase chain reaction analysis for Oct-4 and DAZL gene expression with G6PD gene as internal standard.
G6PD and Oct-4 mRNA was detected in the granulosa cells in 47.6% (10/21). The median of Oct-4 mRNA/G6PD mRNA was 1.75 with intra-quarteral range from 0.10 to 98.21. The OCT-4 mRNA expression was statistically significantly correlated with the number of oocytes retrieved; when the Oct-4 mRNA expression was higher, then more than six oocytes were retrieved (p=0.037, Wilcoxon rank-sum). No detection of DAZL mRNA was found in granulosa cells. There was no additional statistically significant correlation between the levels of Oct-4 expression and FSH basal levels or estradiol peak levels or dosage of FSH for ovulation induction. No association was found between the presence or absence of Oct-4 mRNA expression in granulosa cells and ovarian response to gonadotropin stimulation. Also, no influence on pregnancy was observed between the presence or absence of Oct-4 mRNA expression in granulosa cells or to its expression levels accordingly.
Expression of OCT-4 mRNA, which is a typical stem cell marker and absence of expression of DAZL mRNA, which is a typical germ cell marker, suggest that a subpopulation of luteinized granulosa cells in healthy ovarian follicles (47.6%) consists of stem cells, which are not originated from primordial germ cells. Absence of Oct-4 gene expression in more than half of the cases means probably the end of the productive journey of these cells, towards the oocyte.
Journal of Ovarian Research 11/2012; 5(1):36. DOI:10.1186/1757-2215-5-36 · 2.43 Impact Factor
Available from: Marek Jankowski
- "These experiments led to the observations that OT and its receptor (OTR) are synthesized in the human and rat heart (Gutkowska et al., 1997; Jankowski et al., 1998) and that OT exerts cardio-protection either directly or via stimulation of mediators such as the natriuretic peptides (NPs) (Gutkowska et al., 1997; Gutkowska et al., 2000) and nitric oxide (NO) (Danalache et al., 2007). Since the study showing that OT induces differentiation of EC P19 cells into the functional cardiac muscle (Paquin et al., 2002), several reports have confirmed OT-stimulated cardiomyogenesis in different lines of embryonic stem cells (Fathi et al., 2009; Gassanov et al., 2008a; Hatami et al., 2007; Jankowski et al., 2004; Stefanidis et al., 2009; Uchida et al., 2007). In fact, OT can be considered as an established myogenic morphogen (Breton et al., 2002). "
Embryonic Stem Cells: The Hormonal Regulation of Pluripotency and Embryogenesis, 04/2011; , ISBN: 978-953-307-196-1
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