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Transcriptional regulation induced by cAMP elevation in mouse Schwann cells.

Schmid D, Zeis T, Schaeren-Wiemers N - ASN Neuro (2014)

Bottom Line: One of the strongest differentially regulated gene transcripts was the transcription factor Olig1 (oligodendrocyte transcription factor 1), whose mRNA expression levels were reduced in treated Schwann cells.Olig1 protein was localized in myelinating and nonmyelinating Schwann cells within the sciatic nerve as well as in primary Schwann cells, proposing it as a novel transcription factor of the Schwann cell lineage.Data analysis further revealed that a number of differentially expressed genes in forskolin-treated Schwann cells were associated with the ECM (extracellular matrix), underlining its importance during Schwann cell differentiation in vitro.

View Article: PubMed Central - PubMed

Affiliation: *Neurobiology, Department of Biomedicine, University Hospital Basel, University of Basel, Hebelstrasse 20, CH-4031 Basel, Switzerland.

ABSTRACT
In peripheral nerves, Schwann cell development is regulated by a variety of signals. Some of the aspects of Schwann cell differentiation can be reproduced in vitro in response to forskolin, an adenylyl cyclase activator elevating intracellular cAMP levels. Herein, the effect of forskolin treatment was investigated by a comprehensive genome-wide expression study on primary mouse Schwann cell cultures. Additional to myelin-related genes, many so far unconsidered genes were ascertained to be modulated by forskolin. One of the strongest differentially regulated gene transcripts was the transcription factor Olig1 (oligodendrocyte transcription factor 1), whose mRNA expression levels were reduced in treated Schwann cells. Olig1 protein was localized in myelinating and nonmyelinating Schwann cells within the sciatic nerve as well as in primary Schwann cells, proposing it as a novel transcription factor of the Schwann cell lineage. Data analysis further revealed that a number of differentially expressed genes in forskolin-treated Schwann cells were associated with the ECM (extracellular matrix), underlining its importance during Schwann cell differentiation in vitro. Comparison of samples derived from postnatal sciatic nerves and from both treated and untreated Schwann cell cultures showed considerable differences in gene expression between in vivo and in vitro, allowing us to separate Schwann cell autonomous from tissue-related changes. The whole data set of the cell culture microarray study is provided to offer an interactive search tool for genes of interest.

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Differential gene expression upon forskolin treatment                        The differential expression of the strongest increased (A) and decreased                            (B) gene transcripts was validated by qRT–PCR in treated                            compared with untreated primary mouse Schwann cells. Data were                            normalized to the expression of 60s. The columns represent the mean                            value of 12 experimental samples, and the error bars indicate the S.D.                            *: P≤0.005, **:                            P≤0.0001, ***:                            P≤0.00001. Raw data are provided as                            Supplementary Table S3 (available at http://www.asnneuro.org/an/006/an006e142add.htm).
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Figure 1: Differential gene expression upon forskolin treatment The differential expression of the strongest increased (A) and decreased (B) gene transcripts was validated by qRT–PCR in treated compared with untreated primary mouse Schwann cells. Data were normalized to the expression of 60s. The columns represent the mean value of 12 experimental samples, and the error bars indicate the S.D. *: P≤0.005, **: P≤0.0001, ***: P≤0.00001. Raw data are provided as Supplementary Table S3 (available at http://www.asnneuro.org/an/006/an006e142add.htm).

Mentions: To further investigate the effect of forskolin on transcriptional regulation in cultured mouse Schwann cells, microarray data were analyzed more stringently using an FDR-adjusted P-value of <0.05. This study revealed that forskolin treatment resulted in increased expression of 330 transcripts by at least 1.5-fold, and in decreased expression of 305 transcripts (Supplementary Table S2; available at http://www.asnneuro.org/an/006/an006e142add.htm). Among the 25 strongest induced genes, Mpz was the solely typical so far known myelin-related gene (Table 2A). Strong transcriptional induction was detected for the sclerostin domain containing 1 (Sostdc1), the pleckstrin homology domain containing family A (Plekha4) and the ECM (extracellular matrix) protein spondin 2 (Spon2). Furthermore, increased transcription was detected for the transcription factor Egr3, as already stated before (Table 1A), the fibro-blast growth factor 7 (Fgf7), the endothelin receptor type B (Ednrb) and the proteoglycan decorin (Dcn), to name a few. Strongest down-regulation upon forskolin treatment was detected for the mRNA expression levels of protocadherin 20 (Pcdh20, also known as Pcdh13), phosphodiesterase 1B (Pde1b) and leucine-rich repeats and transmembrane domains 1 (Lrtm1), each represented by two transcripts (Table 2B). Olig1 was the only transcription factor identified among the 25 strongest reduced transcripts. Strong transcriptional reduction was also identified for the chondroitin sulfate proteoglycan 4 [Cspg4, better known as neuron-glial antigen 2 (NG2)], the proteoglycan aggrecan (Acan), the secreted matrix Gla protein (Mgp) and the platelet-derived growth factor subunit B (Pdgfb). qRT–PCR of these highly differentially expressed transcripts validated our microarray for the one which were significantly increased upon forskolin treatment (Figure 1A) as well as for the ones which were down-regulated upon forskolin treatment with the sole exception of Nme7 (Figure 1B).


Transcriptional regulation induced by cAMP elevation in mouse Schwann cells.

Schmid D, Zeis T, Schaeren-Wiemers N - ASN Neuro (2014)

Differential gene expression upon forskolin treatment                        The differential expression of the strongest increased (A) and decreased                            (B) gene transcripts was validated by qRT–PCR in treated                            compared with untreated primary mouse Schwann cells. Data were                            normalized to the expression of 60s. The columns represent the mean                            value of 12 experimental samples, and the error bars indicate the S.D.                            *: P≤0.005, **:                            P≤0.0001, ***:                            P≤0.00001. Raw data are provided as                            Supplementary Table S3 (available at http://www.asnneuro.org/an/006/an006e142add.htm).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4834722&req=5

Figure 1: Differential gene expression upon forskolin treatment The differential expression of the strongest increased (A) and decreased (B) gene transcripts was validated by qRT–PCR in treated compared with untreated primary mouse Schwann cells. Data were normalized to the expression of 60s. The columns represent the mean value of 12 experimental samples, and the error bars indicate the S.D. *: P≤0.005, **: P≤0.0001, ***: P≤0.00001. Raw data are provided as Supplementary Table S3 (available at http://www.asnneuro.org/an/006/an006e142add.htm).
Mentions: To further investigate the effect of forskolin on transcriptional regulation in cultured mouse Schwann cells, microarray data were analyzed more stringently using an FDR-adjusted P-value of <0.05. This study revealed that forskolin treatment resulted in increased expression of 330 transcripts by at least 1.5-fold, and in decreased expression of 305 transcripts (Supplementary Table S2; available at http://www.asnneuro.org/an/006/an006e142add.htm). Among the 25 strongest induced genes, Mpz was the solely typical so far known myelin-related gene (Table 2A). Strong transcriptional induction was detected for the sclerostin domain containing 1 (Sostdc1), the pleckstrin homology domain containing family A (Plekha4) and the ECM (extracellular matrix) protein spondin 2 (Spon2). Furthermore, increased transcription was detected for the transcription factor Egr3, as already stated before (Table 1A), the fibro-blast growth factor 7 (Fgf7), the endothelin receptor type B (Ednrb) and the proteoglycan decorin (Dcn), to name a few. Strongest down-regulation upon forskolin treatment was detected for the mRNA expression levels of protocadherin 20 (Pcdh20, also known as Pcdh13), phosphodiesterase 1B (Pde1b) and leucine-rich repeats and transmembrane domains 1 (Lrtm1), each represented by two transcripts (Table 2B). Olig1 was the only transcription factor identified among the 25 strongest reduced transcripts. Strong transcriptional reduction was also identified for the chondroitin sulfate proteoglycan 4 [Cspg4, better known as neuron-glial antigen 2 (NG2)], the proteoglycan aggrecan (Acan), the secreted matrix Gla protein (Mgp) and the platelet-derived growth factor subunit B (Pdgfb). qRT–PCR of these highly differentially expressed transcripts validated our microarray for the one which were significantly increased upon forskolin treatment (Figure 1A) as well as for the ones which were down-regulated upon forskolin treatment with the sole exception of Nme7 (Figure 1B).

Bottom Line: One of the strongest differentially regulated gene transcripts was the transcription factor Olig1 (oligodendrocyte transcription factor 1), whose mRNA expression levels were reduced in treated Schwann cells.Olig1 protein was localized in myelinating and nonmyelinating Schwann cells within the sciatic nerve as well as in primary Schwann cells, proposing it as a novel transcription factor of the Schwann cell lineage.Data analysis further revealed that a number of differentially expressed genes in forskolin-treated Schwann cells were associated with the ECM (extracellular matrix), underlining its importance during Schwann cell differentiation in vitro.

View Article: PubMed Central - PubMed

Affiliation: *Neurobiology, Department of Biomedicine, University Hospital Basel, University of Basel, Hebelstrasse 20, CH-4031 Basel, Switzerland.

ABSTRACT
In peripheral nerves, Schwann cell development is regulated by a variety of signals. Some of the aspects of Schwann cell differentiation can be reproduced in vitro in response to forskolin, an adenylyl cyclase activator elevating intracellular cAMP levels. Herein, the effect of forskolin treatment was investigated by a comprehensive genome-wide expression study on primary mouse Schwann cell cultures. Additional to myelin-related genes, many so far unconsidered genes were ascertained to be modulated by forskolin. One of the strongest differentially regulated gene transcripts was the transcription factor Olig1 (oligodendrocyte transcription factor 1), whose mRNA expression levels were reduced in treated Schwann cells. Olig1 protein was localized in myelinating and nonmyelinating Schwann cells within the sciatic nerve as well as in primary Schwann cells, proposing it as a novel transcription factor of the Schwann cell lineage. Data analysis further revealed that a number of differentially expressed genes in forskolin-treated Schwann cells were associated with the ECM (extracellular matrix), underlining its importance during Schwann cell differentiation in vitro. Comparison of samples derived from postnatal sciatic nerves and from both treated and untreated Schwann cell cultures showed considerable differences in gene expression between in vivo and in vitro, allowing us to separate Schwann cell autonomous from tissue-related changes. The whole data set of the cell culture microarray study is provided to offer an interactive search tool for genes of interest.

Show MeSH