Ectopic Expression of Single Transcription Factors Directs Differentiation of a Medaka Spermatogonial Cell Line

Thoma, Eva C.; Wagner, Toni U.; Weber, Isabell P.; Herpin, Amaury; Fischer, Andreas; Schartl, Manfred

doi:10.1089/scd.2010.0290

Cited by 19 publications

(17 citation statements)

References 64 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This idea is supported by a similar observation reported in another study describing cell conversion of medakafish spermatogonia into various somatic cell types by ectopic expression of lineage-specific TFs [38]. In that study, the activation of genes that in vivo are located upstream of the inducing factors was also detected for three different processes of cell fate conversion.…”

Section: Discussionsupporting

confidence: 87%

Ectopic Expression of Neurogenin 2 Alone is Sufficient to Induce Differentiation of Embryonic Stem Cells into Mature Neurons

et al. 2012

Self Cite

View full text Add to dashboard Cite

Recent studies show that combinations of defined key developmental transcription factors (TFs) can reprogram somatic cells to pluripotency or induce cell conversion of one somatic cell type to another. However, it is not clear if single genes can define a cell̀s identity and if the cell fate defining potential of TFs is also operative in pluripotent stem cells in vitro. Here, we show that ectopic expression of the neural TF Neurogenin2 (Ngn2) is sufficient to induce rapid and efficient differentiation of embryonic stem cells (ESCs) into mature glutamatergic neurons. Ngn2-induced neuronal differentiation did not require any additional external or internal factors and occurred even under pluripotency-promoting conditions. Differentiated cells displayed neuron-specific morphology, protein expression, and functional features, most importantly the generation of action potentials and contacts with hippocampal neurons. Gene expression analyses revealed that Ngn2-induced in vitro differentiation partially resembled neurogenesis in vivo, as it included specific activation of Ngn2 target genes and interaction partners. These findings demonstrate that a single gene is sufficient to determine cell fate decisions of uncommitted stem cells thus giving insights into the role of key developmental genes during lineage commitment. Furthermore, we present a promising tool to improve directed differentiation strategies for applications in both stem cell research and regenerative medicine.

show abstract

Section: Discussionsupporting

confidence: 87%

Ectopic Expression of Neurogenin 2 Alone is Sufficient to Induce Differentiation of Embryonic Stem Cells into Mature Neurons

et al. 2012

Self Cite

View full text Add to dashboard Cite

show abstract

“…The cell line mes1 was chosen to represent Medaka embryonic stem cells [17], [19] and sg3 to represent adult spermatogonial stem cells [18] with near-pluripotent differentiation potential [20]. We wanted to identify activating transcription factors that were co-regulated in the pluripotent cell lines mes1 and sg3.…”

Section: Resultsmentioning

confidence: 99%

“…Subsequently, the results were used in directed differentiation assays: factors were cloned into expression vectors, transfected into both cell lines and assayed for differentiation. For example, the second hit, MITF, led to successful targeted differentiation of both stem cell lines after ectopic expression of this transcription factor [20].…”

Section: Resultsmentioning

confidence: 99%

CrossQuery: A Web Tool for Easy Associative Querying of Transcriptome Data

et al. 2011

Self Cite

View full text Add to dashboard Cite

Enormous amounts of data are being generated by modern methods such as transcriptome or exome sequencing and microarray profiling. Primary analyses such as quality control, normalization, statistics and mapping are highly complex and need to be performed by specialists. Thereafter, results are handed back to biomedical researchers, who are then confronted with complicated data lists. For rather simple tasks like data filtering, sorting and cross-association there is a need for new tools which can be used by non-specialists. Here, we describe CrossQuery, a web tool that enables straight forward, simple syntax queries to be executed on transcriptome sequencing and microarray datasets. We provide deep-sequencing data sets of stem cell lines derived from the model fish Medaka and microarray data of human endothelial cells. In the example datasets provided, mRNA expression levels, gene, transcript and sample identification numbers, GO-terms and gene descriptions can be freely correlated, filtered and sorted. Queries can be saved for later reuse and results can be exported to standard formats that allow copy-and-paste to all widespread data visualization tools such as Microsoft Excel. CrossQuery enables researchers to quickly and freely work with transcriptome and microarray data sets requiring only minimal computer skills. Furthermore, CrossQuery allows growing association of multiple datasets as long as at least one common point of correlated information, such as transcript identification numbers or GO-terms, is shared between samples. For advanced users, the object-oriented plug-in and event-driven code design of both server-side and client-side scripts allow easy addition of new features, data sources and data types.

show abstract

“…We used pXmitf as effector, a construct expressing the Xiphophorus mitfm, because Xmitf has been well-documented [61] and proven to be efficient to induce melanocyte in MES1 [49] and SG3 cells [62]. Upon cotransfection with pXmitf, the activity increased by 3.6, 3.8 and 4.4 fold in SG3 cells (Figure 2A), by 5.9, 6 and 5.5 in MES1 cells ( Figure 2B) fold for DAZ, DND and VAS, respectively.…”

Section: Ra Suppresses Mitf-activated Germ Gene Transcription In Stemmentioning

confidence: 99%

Retinoic Acid Regulates Germ Gene Transcription In Vitro and Spermatogenesis in Testicular Organ Culture

Hóng¹,

Li²

2012

Biol Syst Open Access

View full text Add to dashboard Cite

The adult testis is the male reproductive organ that produces sex hormones for sexual behaviours and sperm in a well-controlled process called spermatogenesis. Spermatogenesis involves self-renewal and differentiation of the male germ stem cells spermatogonia, culminating in the production of sperm for germline transmission. Retinoic acid (RA) is essential for fate decision and meiosis entry of mammalian germ cells. The microphthalmia-associated transcription factor (Mitf) is a newly identified transcriptional activator of germ genes in the fish medaka (Oryzias latipes). Here we report an essential role of RA on Mitf-activated germ gene expression and spermatogenesis. Medaka germ gene promoters DAZ, DND and VAS were found to contain RA-responsive elements and to exhibit little Mitf-activated transcriptional activity in response to RA treatment in both medaka ES cells and male germ stem cells. A procedure for testicular organ culture was established, which allows for test-tube spermatogenesis over a 21-day period. By the ordered testicular architecture, cellular morphology and VAS-driven GFP expression, different spermatogenic stages were identifiable on testicular cross-sections. In organ culture, RA treatment generated testes completely free of spermatogonia, and reduced VAS-driven GFP expression in spermatogonia but enhanced its expression in meiotic and post-meiotic germ cells. Therefore, RA represses Mitf-activated germ gene expression in stem cell cultures and differentially regulates germ gene expression depending upon the stages of spermatogenesis, and represses spermatogonial stem cell maintenance in cultured testes of medaka as a lower vertebrate model. These results demonstrate that RA plays a highly conserved role in germ cell development from fish to mammals.

show abstract

Ectopic Expression of Single Transcription Factors Directs Differentiation of a Medaka Spermatogonial Cell Line

Cited by 19 publications

References 64 publications

Ectopic Expression of Neurogenin 2 Alone is Sufficient to Induce Differentiation of Embryonic Stem Cells into Mature Neurons

Ectopic Expression of Neurogenin 2 Alone is Sufficient to Induce Differentiation of Embryonic Stem Cells into Mature Neurons

CrossQuery: A Web Tool for Easy Associative Querying of Transcriptome Data

Retinoic Acid Regulates Germ Gene Transcription In Vitro and Spermatogenesis in Testicular Organ Culture

Contact Info

Product

Resources

About