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Leucine-rich repeat kinase 2 modulates retinoic acid-induced neuronal differentiation of murine embryonic stem cells.

Schulz C, Paus M, Frey K, Schmid R, Kohl Z, Mennerich D, Winkler J, Gillardon F - PLoS ONE (2011)

Bottom Line: LRRK2 is expressed in neural precursor cells suggesting a role in neurodevelopment.By contrast, expression of neurotransmitter receptors and neurotransmitter release was increased in LRRK2+/- cultures indicating that LRRK2 promotes neuronal differentiation.Alterations in phosphorylation of the putative LRRK2 substrates, translation initiation factor 4E binding protein 1 and moesin, do not appear to be involved in altered differentiation, rather there is indirect evidence that a regulatory signaling network comprising retinoic acid receptors, let-7 miRNA and downstream target genes/mRNAs may be affected in LRRK2-deficient stem cells in culture.

View Article: PubMed Central - PubMed

Affiliation: Boehringer Ingelheim Pharma GmbH & Co KG, CNS Research, Biberach an der Riss, Germany.

ABSTRACT

Background: Dominant mutations in the leucine-rich repeat kinase 2 (LRRK2) gene are the most prevalent cause of Parkinson's disease, however, little is known about the biological function of LRRK2 protein. LRRK2 is expressed in neural precursor cells suggesting a role in neurodevelopment.

Methodology/principal findings: In the present study, differential gene expression profiling revealed a faster silencing of pluripotency-associated genes, like Nanog, Oct4, and Lin28, during retinoic acid-induced neuronal differentiation of LRRK2-deficient mouse embryonic stem cells compared to wildtype cultures. By contrast, expression of neurotransmitter receptors and neurotransmitter release was increased in LRRK2+/- cultures indicating that LRRK2 promotes neuronal differentiation. Consistently, the number of neural progenitor cells was higher in the hippocampal dentate gyrus of adult LRRK2-deficient mice. Alterations in phosphorylation of the putative LRRK2 substrates, translation initiation factor 4E binding protein 1 and moesin, do not appear to be involved in altered differentiation, rather there is indirect evidence that a regulatory signaling network comprising retinoic acid receptors, let-7 miRNA and downstream target genes/mRNAs may be affected in LRRK2-deficient stem cells in culture.

Conclusion/significance: Parkinson's disease-linked LRRK2 mutations that associated with enhanced kinase activity may affect retinoic acid receptor signaling during neurodevelopment and/or neuronal maintenance as has been shown in other mouse models of chronic neurodegenerative diseases.

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Related in: MedlinePlus

Affymetrix GeneChip analysis comparing mRNA expression levels in: (A) clone A LRRK2+/− ES cells versus wildtype ES cells, (C) clone A LRRK2+/− ES cell-derived neurons versus wildtype ES cell-derived neurons, (D) clone B LRRK2+/− ES cell-derived neurons versus wildtype ES cell-derived neurons, and (B) clone A LRRK2+/− ES cell-derived neurons versus clone B LRRK2+/− ES cell-derived neurons.Massive changes in gene expression become detectable in LRRK2+/− ES cell-derived neurons compared to wildtype ES cell-derived neurons 7 days following neuronal differentiation by retinoic acid administration.
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pone-0020820-g003: Affymetrix GeneChip analysis comparing mRNA expression levels in: (A) clone A LRRK2+/− ES cells versus wildtype ES cells, (C) clone A LRRK2+/− ES cell-derived neurons versus wildtype ES cell-derived neurons, (D) clone B LRRK2+/− ES cell-derived neurons versus wildtype ES cell-derived neurons, and (B) clone A LRRK2+/− ES cell-derived neurons versus clone B LRRK2+/− ES cell-derived neurons.Massive changes in gene expression become detectable in LRRK2+/− ES cell-derived neurons compared to wildtype ES cell-derived neurons 7 days following neuronal differentiation by retinoic acid administration.

Mentions: To get more insight into the cellular function of LRRK2, we performed differential gene expression profiling using RNA samples from undifferentiated, SSEA1-immunopositive ES cells and from differentiated, β-III tubulin-immunoreactive ES cell-derived neurons, respectively. As shown in Figure 3, only 9 genes were significantly (fold change ≥1.5 and adjusted p-value≤0.01) down-regulated and 14 genes were up-regulated in LRRK2+/− ES cells compared to wildtype controls. By contrast, 7 days after starting retinoic acid-induced differentiation, 2404 genes were significantly down-regulated and 575 genes were up-regulated in LRRK2+/− ES cell-derived neurons compared to wildtype cultures. Similar changes were detected when two clones (A, B) with monoallelic LRRK2 deletion were compared to wildtype ES cell-derived neurons, whereas gene expression did not differ between the LRRK2+/− clones A and B (Figure 3). The complete Genechip dataset has been loaded into ArrayExpress database (www.ebi.ac.uk/aerep/login) under experiment E-MEXP-2963. These findings indicate that LRRK2 adopts a critical biological function during retinoic acid-induced neuronal differentiation, although LRRK2 is already expressed at the ES cell stage.


Leucine-rich repeat kinase 2 modulates retinoic acid-induced neuronal differentiation of murine embryonic stem cells.

Schulz C, Paus M, Frey K, Schmid R, Kohl Z, Mennerich D, Winkler J, Gillardon F - PLoS ONE (2011)

Affymetrix GeneChip analysis comparing mRNA expression levels in: (A) clone A LRRK2+/− ES cells versus wildtype ES cells, (C) clone A LRRK2+/− ES cell-derived neurons versus wildtype ES cell-derived neurons, (D) clone B LRRK2+/− ES cell-derived neurons versus wildtype ES cell-derived neurons, and (B) clone A LRRK2+/− ES cell-derived neurons versus clone B LRRK2+/− ES cell-derived neurons.Massive changes in gene expression become detectable in LRRK2+/− ES cell-derived neurons compared to wildtype ES cell-derived neurons 7 days following neuronal differentiation by retinoic acid administration.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3111438&req=5

pone-0020820-g003: Affymetrix GeneChip analysis comparing mRNA expression levels in: (A) clone A LRRK2+/− ES cells versus wildtype ES cells, (C) clone A LRRK2+/− ES cell-derived neurons versus wildtype ES cell-derived neurons, (D) clone B LRRK2+/− ES cell-derived neurons versus wildtype ES cell-derived neurons, and (B) clone A LRRK2+/− ES cell-derived neurons versus clone B LRRK2+/− ES cell-derived neurons.Massive changes in gene expression become detectable in LRRK2+/− ES cell-derived neurons compared to wildtype ES cell-derived neurons 7 days following neuronal differentiation by retinoic acid administration.
Mentions: To get more insight into the cellular function of LRRK2, we performed differential gene expression profiling using RNA samples from undifferentiated, SSEA1-immunopositive ES cells and from differentiated, β-III tubulin-immunoreactive ES cell-derived neurons, respectively. As shown in Figure 3, only 9 genes were significantly (fold change ≥1.5 and adjusted p-value≤0.01) down-regulated and 14 genes were up-regulated in LRRK2+/− ES cells compared to wildtype controls. By contrast, 7 days after starting retinoic acid-induced differentiation, 2404 genes were significantly down-regulated and 575 genes were up-regulated in LRRK2+/− ES cell-derived neurons compared to wildtype cultures. Similar changes were detected when two clones (A, B) with monoallelic LRRK2 deletion were compared to wildtype ES cell-derived neurons, whereas gene expression did not differ between the LRRK2+/− clones A and B (Figure 3). The complete Genechip dataset has been loaded into ArrayExpress database (www.ebi.ac.uk/aerep/login) under experiment E-MEXP-2963. These findings indicate that LRRK2 adopts a critical biological function during retinoic acid-induced neuronal differentiation, although LRRK2 is already expressed at the ES cell stage.

Bottom Line: LRRK2 is expressed in neural precursor cells suggesting a role in neurodevelopment.By contrast, expression of neurotransmitter receptors and neurotransmitter release was increased in LRRK2+/- cultures indicating that LRRK2 promotes neuronal differentiation.Alterations in phosphorylation of the putative LRRK2 substrates, translation initiation factor 4E binding protein 1 and moesin, do not appear to be involved in altered differentiation, rather there is indirect evidence that a regulatory signaling network comprising retinoic acid receptors, let-7 miRNA and downstream target genes/mRNAs may be affected in LRRK2-deficient stem cells in culture.

View Article: PubMed Central - PubMed

Affiliation: Boehringer Ingelheim Pharma GmbH & Co KG, CNS Research, Biberach an der Riss, Germany.

ABSTRACT

Background: Dominant mutations in the leucine-rich repeat kinase 2 (LRRK2) gene are the most prevalent cause of Parkinson's disease, however, little is known about the biological function of LRRK2 protein. LRRK2 is expressed in neural precursor cells suggesting a role in neurodevelopment.

Methodology/principal findings: In the present study, differential gene expression profiling revealed a faster silencing of pluripotency-associated genes, like Nanog, Oct4, and Lin28, during retinoic acid-induced neuronal differentiation of LRRK2-deficient mouse embryonic stem cells compared to wildtype cultures. By contrast, expression of neurotransmitter receptors and neurotransmitter release was increased in LRRK2+/- cultures indicating that LRRK2 promotes neuronal differentiation. Consistently, the number of neural progenitor cells was higher in the hippocampal dentate gyrus of adult LRRK2-deficient mice. Alterations in phosphorylation of the putative LRRK2 substrates, translation initiation factor 4E binding protein 1 and moesin, do not appear to be involved in altered differentiation, rather there is indirect evidence that a regulatory signaling network comprising retinoic acid receptors, let-7 miRNA and downstream target genes/mRNAs may be affected in LRRK2-deficient stem cells in culture.

Conclusion/significance: Parkinson's disease-linked LRRK2 mutations that associated with enhanced kinase activity may affect retinoic acid receptor signaling during neurodevelopment and/or neuronal maintenance as has been shown in other mouse models of chronic neurodegenerative diseases.

Show MeSH
Related in: MedlinePlus