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. 2011 Mar-Apr;11(3-4):285-98.
doi: 10.1016/j.gep.2011.02.001. Epub 2011 Feb 15.

Ontological aspects of pluripotency and stemness gene expression pattern in the rhesus monkey

Namdori R Mtango  1 , Catherine A VandeVoort, Keith E Latham
Affiliations

Ontological aspects of pluripotency and stemness gene expression pattern in the rhesus monkey

Namdori R Mtango et al. Gene Expr Patterns. 2011 Mar-Apr.

Abstract

Two essential aspects of mammalian development are the progressive specialization of cells toward different lineages, and the maintenance of progenitor cells that will give rise to the differentiated components of each tissue and also contribute new cells as older cells die or become injured. The transition from totipotentiality to pluripotentiality, to multipotentiality, to monopotentiality, and then to differentiation is a continuous process during development. The ontological relationship between these different stages is not well understood. We report for the first time an ontological survey of expression of 45 putative "stemness" and "pluripotency" genes in rhesus monkey oocytes and preimplantation stage embryos, and comparison to the expression in the inner cell mass, trophoblast stem cells, and a rhesus monkey (ORMES6) embryonic stem cell line. Our results reveal that some of these genes are not highly expressed in all totipotent or pluripotent cell types. Some are predominantly maternal mRNAs present in oocytes and embryos before transcriptional activation, and diminishing before the blastocyst stage. Others are well expressed in morulae or early blastocysts, but are poorly expressed in later blastocysts or ICMs. Also, some of the genes employed to induce pluripotent stem cells from somatic cells (iPS genes) appear unlikely to play major roles as stemness or pluripotency genes in normal embryos.

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Figures

Figure 1
Figure 1
Schematic summary of different groups of genes analyzed.
Figure 2
Figure 2
Summary of ontogenic comparison during transition from embryos to stem cell. The figure shows comparison in transition between A. Inner cell mass (ICMs) and early outgrowth (EO); B. Early outgrowth and undifferentiated ORMES6; C. undifferentiated ORMES6 and early differentiated ORMES6; D. ICMS and trophoblast stem cells (TSC); E. undifferentiated ORMES6 and TSC.
Figure 3
Figure 3
Temporal expression patterns of genes involved in inducing pluripotency in rhesus monkey oocytes, embryos and different stem cell lines. A. Monkey graph show the relative levels of expression for GV and MII stage oocytes and pronucleate through hatched blastocyst-stage embryos produced by in vitro fertilization of oocytes from human chorionic gonadotropin (hCG)-stimulated females and then cultured in vitro in HECM9. GV, germinal vesicle stage oocyte; MII, metaphase II stage oocyte; PN, pronucleate 1-cell stage embryo; 2C, 2-cell stage embryo; 8C, 8-cell stage embryo; 8–16C aAm, 8- to 16-cell stage cultured in α-amanitin; EB, early blastocyst; XB, expanded blastocyst; HB, hatched blastocyst. Statistically significant differences in gene expression corresponding to some of the major increases or decreases in expression are denoted by the brackets for comparisons between stages at the ends of the brackets. Letters a–d indicate P <0.05, 0.01, 0.001 and 0.0001, respectively. N/D indicates not detected. Expression data for the mRNAs encoding the indicated proteins are expressed as the mean cpm bound and the SE is indicated.. B. Expression patterns of stem cell transition. ICM, Inner Cell Mass; EO, early outgrowth; ND, non differentiated ORMES 6 (Oregon Rhesus Macaque Embryonic Stem); ED, Early differentiation; Feeder cells from mouse fibroblast. Data are presented as the mean cpm bound and the SE is indicated. Statistically significant differences in gene expression corresponding to some of the major increases or decreases in expression are denoted by the brackets for comparisons between stages at the ends of the brackets. Letters a–d indicate P <0.05, 0.01, 0.001 and 0.0001, respectively. N/D indicates not detected. The lines drawn between graphs shows how the mRNA levels were reduced or increased between embryos and the transition to stem cells.
Figure 4
Figure 4
Temporal expression patterns of mRNAs encoding genes involved in maintaining pluripotency, self-renewal, growth regulation in rhesus monkey oocytes, embryos and a transition to stem cells. Data are presented as in Fig. 3.
Figure 5
Figure 5
Temporal expression patterns of mRNAs encoding ecto-, meso- and endoderm markers in rhesus monkey oocytes, embryos and a transition to stem cells. Data are presented as in Fig. 3.
Figure 6
Figure 6
Temporal expression patterns of mRNAs involved in early differentiation markers in rhesus monkey oocytes, embryos and a transition to stem cells. Data are presented as in Fig. 3.
Figure 7
Figure 7
Temporal expression patterns of trophoblast markers mRNAs in rhesus monkey oocytes, embryos and a transition to stem cells. Data are presented as in Fig. 3.
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