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Reverse transcription-polymerase chain reaction (RT-PCR) analysis of the selected genes after microarray analysis of the Lin28 knocked down mouse embryonic stem cells (mESCs). The expression of target genes selected by statistical method and filtering procedure after microarray analysis was examined by RT-PCR using specific primers with cDNA from control and Lin28 knocked down mESCs. Microphotographs representative of the results of (A) up-regulated and (B) down-regulated genes after RT-PCR are shown. Gapdh was used as a loading control. siCon, control siRNA transfected; siLin28, Lin28 siRNA transfected.
Source publication
Lin28 has been known to control the proliferation and pluripotency of embryonic stem cells. The purpose of this study was to determine the downstream effectors of Lin28 in mouse embryonic stem cells (mESCs) by RNA interference and microarray analysis.
The control siRNA and Lin28 siRNA (Dharmacon) were transfected into mESCs. Total RNA was prepared...
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Background:
In Ewing sarcoma (EwS), long-term treatment effects and poor survival rates for relapsed or metastatic cases require individualization of therapy and the discovery of new treatment methods. Tumor glucose metabolic activity varies significantly between patients, and FDG-PET signals have been proposed as prognostic factors. However, the...
Citations
... Obox1 was found exclusively expressed in mouse oocytes as early as one-layer follicles and throughout folliculogenesis (Rajkovic et al., 2002). In mouse stem cells, Obox genes were negatively regulated by Lin28 (Park et al., 2012). CPEB, a sequence-specific RNA binding protein, binds to Obox1 mRNA and may regulate its polyadenylation-induced translation (Racki and Richter, 2006). ...
... Obox1 was found exclusively expressed in mouse oocytes as early as one-layer follicles and throughout folliculogenesis (Rajkovic et al., 2002). In mouse stem cells, Obox genes were negatively regulated by Lin28 (Park et al., 2012). CPEB, a sequence-specific RNA binding protein, binds to Obox1 mRNA and may regulate its polyadenylation-induced translation (Racki and Richter, 2006). ...
Mammalian oocytes possess fascinating unknown factors, which can reprogram terminally differentiated germ cells or somatic cells into totipotent embryos. Here, we demonstrate that oocyte-specific homeobox 1 (Obox1), an oocyte-specific factor, can markedly enhance the generation of induced pluripotent stem cells (iPSCs) from mouse fibroblasts in a proliferation-independent manner and can replace Sox2 to achieve pluripotency. Overexpression of Obox1 can greatly promote mesenchymal-to-epithelial transition (MET) at early stage of OSKM-induced reprogramming, and meanwhile, the hyperproliferation of THY1-positive cells can be significantly mitigated. Subsequently, the proportion of THY1-negative cells and Oct4-GFP-positive cells increased dramatically. Further analysis of gene expression and targets of Obox1 during reprogramming indicates that the expression of Obox1 can promote epithelial gene expression and modulate cell-cycle-related gene expression. Taken together, we conclude that the oocyte-specific factor Obox1 serves as a strong activator for somatic cell reprogramming through promoting the MET and mitigating cell hyperproliferation.
... Analysis of putative target genes of LIN28 against the list established as common targets for Pou5f1 and Nanog in human embryonic stem cells (Hosseinpour et al., 2013;Park et al., 2012) revealed that in the testicular RNA-seq data some of them were specifically altered in Fxra À/À males compared with WT males ( Figure 7F) with 50% upregulated (Pou5f1; Nanog, Utf1) and 50% downregulated (Tiam1, Cdh2). ...
... Analysis of putative target genes of LIN28 against the list established as common targets for Pou5f1 and Nanog in human embryonic stem cells (Hosseinpour et al., 2013;Park et al., 2012) revealed that in the testicular RNA-seq data some of them were specifically altered in Fxra À/À males compared with WT males ( Figure 7F) with 50% upregulated (Pou5f1; Nanog, Utf1) and 50% downregulated (Tiam1, Cdh2). ...
Spermatogenesis is the process by which spermatozoa are generated from spermatogonia. This cell population is heterogeneous, with self-renewing spermatogonial stem cells (SSCs) and progenitor spermatogonia that will continue on a path of differentiation. Only SSCs have the ability to regenerate and sustain spermatogenesis. This makes the testis a good model to investigate stem cell biology. The Farnesoid X Receptor alpha (FXRα) was recently shown to be expressed in the testis. However, its global impact on germ cell homeostasis has not yet been studied. Here, using a phenotyping approach in Fxrα−/− mice, we describe unexpected roles of FXRα on germ cell physiology independent of its effects on somatic cells. FXRα helps establish and maintain an undifferentiated germ cell pool and in turn influences male fertility. FXRα regulates the expression of several pluripotency factors. Among these, in vitro approaches show that FXRα controls the expression of the pluripotency marker Lin28 in the germ cells.
... There are six Obox (oocyte-specific homeobox) family transcripts and Obox-1, 2, 3, and 5 mRNA they have been detected in oocytes from growing primary follicles [67], playing then an important role in early embryogenesis [68,69] by orchestrating gene transcription, either ubiquitously or in a tissue-specific manner. Mice lacking the Obox6 gene grow without morphological abnormalities and with normal fertility, indicating a functional redundancy among the Obox family members [70]. ...
Gene expression is required in all steps of embryonic development and therefore heat stress is known to reduce developmental competence after direct exposure of oocytes and embryos to different conditions of heat shock, by decreasing protein synthesis. Moreover, as in somatic cells, the heat stress befuddles the integration of RNA and posttranscriptional modification of RNA, the assumption was that during meiotic maturation heat shock may mutate RNA within oocytes, with the possibility of altering the surrounding cumulus cells, causing, thus, reductions in development. Heat shock proteins (HSP) are among the first proteins produced during embryonic development and are crucial to cell function. The HSP70 (HSPA14 gene) is an important part of the cell’s machinery for folding, unfolding, transport, localization of proteins and differentiation, regulation of the embryonic cell cycle and helping to protect cells from stress. Therefore, HSPA14 is an apoptotic gene induced by heat shock is associated with embryonic loss, playing an important role of control mechanism of processes involved in growth, cellular differentiation, and embryonic development. In addition the connexin proteins (e.g. Cx43), related to gap junctions, are expressed in numerous tissues including gonads, act as a mediator of heat stress effect on cells. In the present review, the effect of heat stress on bovine embryonic development in a physiologic and genetic point of view is fully discussed.
Protein phosphorylation plays an important role in animal reproduction. However, its role in the onset of puberty in goats remains largely unexplored. Accordingly, in the present study, the molecular changes controlling the onset of puberty in goats were investigated by identifying the differentially phosphorylated proteins (DPPs) and phosphorylation sites (DPSs) in the hypothalami of prepubertal and pubertal female goats using LC–MS/MS and tandem mass tag labelling. A total of 3265 phosphopeptides corresponding to 1628 phosphoproteins were identified, including 234 upregulated and 342 downregulated phosphopeptides. The DPSs HTT, MAP1B, CAMKK1, MAP2, DNAJC5, and GAP43 were identified. These DPPs are enriched in the endocytosis, cAMP signaling, Rap1 signaling, melanogenesis, and insulin secretion pathways. These pathways are related to gonadotropin-releasing hormone and puberty. In particular, glucose-6-phosphate isomerase, fructose-bisphosphate aldolase C, and fructose-bisphosphate aldolase A occupy important locations in the protein-protein interaction network. These data provide evidence for a complex interaction network in goat hypothalamus proteins that affects puberty. Furthermore, they may help identify new puberty-regulating candidates and/or serve as an important resource for exploring the physiological mechanism of puberty onset in mammals.
Significance
This study provides evidence for a complex interaction network in goat hypothalamus proteins that affects puberty. Furthermore, our data may help identify new puberty-regulating candidates and/or serve as an important resource for exploring the physiological mechanism of puberty onset in mammals.
