FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W2111034192> ?p ?o ?g. }

• W2111034192 endingPage "250" @default.
• W2111034192 startingPage "242" @default.
• W2111034192 abstract "•Engrailed regulates DNA damage response and chromatin remodeling•Dopaminergic neurons are protected by Engrailed from oxidative stress•Engrailed acts both at genetic and epigenetic levels to confer neuroprotection•Engrailed homeoprotein transduction has therapeutic potential Engrailed homeoproteins are expressed in adult dopaminergic neurons of the substantia nigra. In Engrailed1 heterozygous mice, these neurons start dying at 6 weeks, are more sensitive to oxidative stress, and progressively develop traits similar to those observed following an acute and strong oxidative stress inflected to wild-type neurons. These changes include DNA strand breaks and the modification (intensity and distribution) of several nuclear and nucleolar heterochromatin marks. Engrailed1 and Engrailed2 are biochemically equivalent transducing proteins previously used to antagonize dopaminergic neuron death in Engrailed1 heterozygous mice and in mouse models of Parkinson disease. Accordingly, we show that, following an acute oxidative stress, a single Engrailed2 injection restores all nuclear and nucleolar heterochromatin marks, decreases the number of DNA strand breaks, and protects dopaminergic neurons against apoptosis. Engrailed homeoproteins are expressed in adult dopaminergic neurons of the substantia nigra. In Engrailed1 heterozygous mice, these neurons start dying at 6 weeks, are more sensitive to oxidative stress, and progressively develop traits similar to those observed following an acute and strong oxidative stress inflected to wild-type neurons. These changes include DNA strand breaks and the modification (intensity and distribution) of several nuclear and nucleolar heterochromatin marks. Engrailed1 and Engrailed2 are biochemically equivalent transducing proteins previously used to antagonize dopaminergic neuron death in Engrailed1 heterozygous mice and in mouse models of Parkinson disease. Accordingly, we show that, following an acute oxidative stress, a single Engrailed2 injection restores all nuclear and nucleolar heterochromatin marks, decreases the number of DNA strand breaks, and protects dopaminergic neurons against apoptosis. Engrailed1 (En1) and Engrailed2 (En2), collectively Engrailed or En1/2 homeoproteins (HPs), play equivalent roles in the survival of adult mesencephalic dopaminergic (mDA) neurons in the substantia nigra pars compacta (SNpc) and the ventral tegmental area (VTA) (Albéri et al., 2004Albéri L. Sgadò P. Simon H.H. Engrailed genes are cell-autonomously required to prevent apoptosis in mesencephalic dopaminergic neurons.Development. 2004; 131: 3229-3236Crossref PubMed Scopus (151) Google Scholar, Sgadò et al., 2006Sgadò P. Albéri L. Gherbassi D. Galasso S.L. Ramakers G.M. Alavian K.N. Smidt M.P. Dyck R.H. Simon H.H. Slow progressive degeneration of nigral dopaminergic neurons in postnatal Engrailed mutant mice.Proc. Natl. Acad. Sci. USA. 2006; 103: 15242-15247Crossref PubMed Scopus (109) Google Scholar). In the En1−/+;En2+/+ (En1+/−) mouse, mDA neurons degenerate progressively starting at 6 weeks (Sonnier et al., 2007Sonnier L. Le Pen G. Hartmann A. Bizot J.C. Trovero F. Krebs M.O. Prochiantz A. Progressive loss of dopaminergic neurons in the ventral midbrain of adult mice heterozygote for Engrailed1.J. Neurosci. 2007; 27: 1063-1071Crossref PubMed Scopus (116) Google Scholar). As in Parkinson disease (PD), death is higher in the SNpc than in the VTA and En1+/− mice display motor and non-motor behaviors (Sonnier et al., 2007Sonnier L. Le Pen G. Hartmann A. Bizot J.C. Trovero F. Krebs M.O. Prochiantz A. Progressive loss of dopaminergic neurons in the ventral midbrain of adult mice heterozygote for Engrailed1.J. Neurosci. 2007; 27: 1063-1071Crossref PubMed Scopus (116) Google Scholar). This and a possible association between EN polymorphisms and the risk to develop PD (Fuchs et al., 2009Fuchs J. Mueller J.C. Lichtner P. Schulte C. Munz M. Berg D. Wüllner U. Illig T. Sharma M. Gasser T. The transcription factor PITX3 is associated with sporadic Parkinson’s disease.Neurobiol. Aging. 2009; 30: 731-738Abstract Full Text Full Text PDF PubMed Scopus (86) Google Scholar, Haubenberger et al., 2011Haubenberger D. Reinthaler E. Mueller J.C. Pirker W. Katzenschlager R. Froehlich R. Bruecke T. Daniel G. Auff E. Zimprich A. Association of transcription factor polymorphisms PITX3 and EN1 with Parkinson’s disease.Neurobiol. Aging. 2011; 32: 302-307Abstract Full Text Full Text PDF PubMed Scopus (60) Google Scholar, Rissling et al., 2009Rissling I. Strauch K. Höft C. Oertel W.H. Möller J.C. Haplotype analysis of the engrailed-2 gene in young-onset Parkinson’s disease.Neurodegener. Dis. 2009; 6: 102-105Crossref PubMed Scopus (14) Google Scholar) suggest that Engrailed might be in the PD pathway. Thanks to HP internalization properties (Joliot and Prochiantz, 2004Joliot A. Prochiantz A. Transduction peptides: from technology to physiology.Nat. Cell Biol. 2004; 6: 189-196Crossref PubMed Scopus (449) Google Scholar), it was shown that Engrailed transduction saves mDA neurons in the En1+/− mouse (Sonnier et al., 2007Sonnier L. Le Pen G. Hartmann A. Bizot J.C. Trovero F. Krebs M.O. Prochiantz A. Progressive loss of dopaminergic neurons in the ventral midbrain of adult mice heterozygote for Engrailed1.J. Neurosci. 2007; 27: 1063-1071Crossref PubMed Scopus (116) Google Scholar) and in wild-type (WT) mice exposed to striatal 6-hydroxydopamine (6-OHDA), systemic 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridin (MPTP), or the toxic A30P variant of alpha-synuclein (Alvarez-Fischer et al., 2011Alvarez-Fischer D. Fuchs J. Castagner F. Stettler O. Massiani-Beaudoin O. Moya K.L. Bouillot C. Oertel W.H. Lombès A. Faigle W. et al.Engrailed protects mouse midbrain dopaminergic neurons against mitochondrial complex I insults.Nat. Neurosci. 2011; 14: 1260-1266Crossref PubMed Scopus (102) Google Scholar). Engrailed survival activity involves its ability to stimulate the translation of mitochondrial complex I subunits Ndufs1 and Ndufs3, resulting in enhanced ATP synthesis (Alvarez-Fischer et al., 2011Alvarez-Fischer D. Fuchs J. Castagner F. Stettler O. Massiani-Beaudoin O. Moya K.L. Bouillot C. Oertel W.H. Lombès A. Faigle W. et al.Engrailed protects mouse midbrain dopaminergic neurons against mitochondrial complex I insults.Nat. Neurosci. 2011; 14: 1260-1266Crossref PubMed Scopus (102) Google Scholar, Stettler et al., 2012Stettler O. Joshi R.L. Wizenmann A. Reingruber J. Holcman D. Bouillot C. Castagner F. Prochiantz A. Moya K.L. Engrailed homeoprotein recruits the adenosine A1 receptor to potentiate ephrin A5 function in retinal growth cones.Development. 2012; 139: 215-224Crossref PubMed Scopus (53) Google Scholar). Indeed, Engrailed is a multifunctional protein regulating the translation of capped mRNAs (Brunet et al., 2005Brunet I. Weinl C. Piper M. Trembleau A. Volovitch M. Harris W. Prochiantz A. Holt C. The transcription factor Engrailed-2 guides retinal axons.Nature. 2005; 438: 94-98Crossref PubMed Scopus (203) Google Scholar). We now find that the loss of one En1 allele leads to changes in the expression of many genes in the DNA damage response (DDR) and chromatin structure pathways, often observed upon oxidative stress (Canugovi et al., 2013Canugovi C. Misiak M. Ferrarelli L.K. Croteau D.L. Bohr V.A. The role of DNA repair in brain related disease pathology.DNA Repair (Amst.). 2013; 12: 578-587Crossref PubMed Scopus (100) Google Scholar), suggesting that Engrailed protects against oxidative stress. Indeed, mDA neurons from En1+/− mice present signs of DNA damage and chromatin alterations and are more sensitive to an acute oxidative stress. Moreover, Engrailed transduction protects against oxidative stress-induced phenotypes. RNA sequencing (RNA-seq) analysis on microdissected SNpc was performed in WT and En1+/− mice. Comparable reads were obtained in WT and En1+/− mice with 989 differentially expressed genes (p < 0.05) (Figure S1A; Table S1). Analysis was performed on 6-week-old animals when all neurons are still present in the in En1+/− mice (Sonnier et al., 2007Sonnier L. Le Pen G. Hartmann A. Bizot J.C. Trovero F. Krebs M.O. Prochiantz A. Progressive loss of dopaminergic neurons in the ventral midbrain of adult mice heterozygote for Engrailed1.J. Neurosci. 2007; 27: 1063-1071Crossref PubMed Scopus (116) Google Scholar). Pathway Studio Ontology (gene set enrichment analysis, Pathway Studio software) indicates that the three most represented and significant groups are DNA repair (p = 0.002), chromatin remodeling (p = 0.004), and transcription factors (p = 0.007); Cell Process Pathways analysis also revealed differential apoptosis regulation genes (p = 0.01) (Figure S1B). Genes within these ontologies and pathways were ranked by increasing p values; Figure 1A highlights those with significant differences in read numbers. Transcription factor genes represent the most abundant group (Figure S1B); Figure S1D ranks by p values those with modified expression in En1+/− mice. En2 infusion in the SNpc of WT mice also modifies transcription factor genes, but much less so for DDR and chromatin-modifying ones (Figure S1C) that were thus further investigated in the context of this study. The qRT-PCR (Figure S1E) confirmed that 6-week-old En1+/− mice display altered expression of several genes related to DNA damage, chromatin remodeling, and apoptosis (Asagoshi et al., 2010Asagoshi K. Tano K. Chastain 2nd, P.D. Adachi N. Sonoda E. Kikuchi K. Koyama H. Nagata K. Kaufman D.G. Takeda S. et al.FEN1 functions in long patch base excision repair under conditions of oxidative stress in vertebrate cells.Mol. Cancer Res. 2010; 8: 204-215Crossref PubMed Scopus (28) Google Scholar, Burikhanov et al., 2014Burikhanov R. Shrestha-Bhattarai T. Hebbar N. Qiu S. Zhao Y. Zambetti G.P. Rangnekar V.M. Paracrine apoptotic effect of p53 mediated by tumor suppressor Par-4.Cell Rep. 2014; 6: 271-277Abstract Full Text Full Text PDF PubMed Scopus (26) Google Scholar, Choi et al., 2004Choi W.S. Eom D.S. Han B.S. Kim W.K. Han B.H. Choi E.J. Oh T.H. Markelonis G.J. Cho J.W. Oh Y.J. Phosphorylation of p38 MAPK induced by oxidative stress is linked to activation of both caspase-8- and -9-mediated apoptotic pathways in dopaminergic neurons.J. Biol. Chem. 2004; 279: 20451-20460Crossref PubMed Scopus (185) Google Scholar, Hong et al., 2008Hong Z. Jiang J. Lan L. Nakajima S. Kanno S. Koseki H. Yasui A. A polycomb group protein, PHF1, is involved in the response to DNA double-strand breaks in human cell.Nucleic Acids Res. 2008; 36: 2939-2947Crossref PubMed Scopus (79) Google Scholar, Ma and D’Mello, 2011Ma C. D’Mello S.R. Neuroprotection by histone deacetylase-7 (HDAC7) occurs by inhibition of c-jun expression through a deacetylase-independent mechanism.J. Biol. Chem. 2011; 286: 4819-4828Crossref PubMed Scopus (65) Google Scholar, Melis et al., 2013Melis J.P. Kuiper R.V. Zwart E. Robinson J. Pennings J.L. van Oostrom C.T. Luijten M. van Steeg H. Slow accumulation of mutations in Xpc-/- mice upon induction of oxidative stress.DNA Repair (Amst.). 2013; 12: 1081-1086Crossref PubMed Scopus (35) Google Scholar, Yang et al., 2013Yang Y. Wang C. Zhang P. Gao K. Wang D. Yu H. Zhang T. Jiang S. Hexige S. Hong Z. et al.Polycomb group protein PHF1 regulates p53-dependent cell growth arrest and apoptosis.J. Biol. Chem. 2013; 288: 529-539Crossref PubMed Scopus (26) Google Scholar, Zhang et al., 2013Zhang X. Lv L. Chen Q. Yuan F. Zhang T. Yang Y. Zhang H. Wang Y. Jia Y. Qian L. et al.Mouse DNA polymerase kappa has a functional role in the repair of DNA strand breaks.DNA Repair (Amst.). 2013; 12: 377-388Crossref PubMed Scopus (26) Google Scholar). The mDA neurons in the SNpc of En1+/− mice (between 6 and 8 weeks of age) were examined for signs of DNA damage by following the DNA strand break marker γ-H2AX (Löbrich et al., 2010Löbrich M. Shibata A. Beucher A. Fisher A. Ensminger M. Goodarzi A.A. Barton O. Jeggo P.A. gammaH2AX foci analysis for monitoring DNA double-strand break repair: strengths, limitations and optimization.Cell Cycle. 2010; 9: 662-669Crossref PubMed Scopus (507) Google Scholar). This revealed the presence of multiple γ-H2AX foci in about 16% of tyrosine hydroxylase-positive (TH+) neurons in the SNpc (Figures 1B and 1C). Of note, DNA strand breaks do not necessarily lead to rapid cell death, as shown by the absence of significant death in the En1+/− mutant between 24 and 48 weeks (Sonnier et al., 2007Sonnier L. Le Pen G. Hartmann A. Bizot J.C. Trovero F. Krebs M.O. Prochiantz A. Progressive loss of dopaminergic neurons in the ventral midbrain of adult mice heterozygote for Engrailed1.J. Neurosci. 2007; 27: 1063-1071Crossref PubMed Scopus (116) Google Scholar), even though 16% of the cells have multiple γ-H2AX foci at 24 weeks (Figure S1F). In WT mice, about 98% of mDA neurons display a single ring of γ-H2AX around the nucleolus (Figure 1B), also present in neurons throughout the brain. In line with the lesser sensitivity of VTA mDA neurons to the loss of one En1 allele (Sonnier et al., 2007Sonnier L. Le Pen G. Hartmann A. Bizot J.C. Trovero F. Krebs M.O. Prochiantz A. Progressive loss of dopaminergic neurons in the ventral midbrain of adult mice heterozygote for Engrailed1.J. Neurosci. 2007; 27: 1063-1071Crossref PubMed Scopus (116) Google Scholar), γ-H2AX staining was similar in the VTA of WT and En1+/− mice (Figure 1C). En1+/− mice also have signs of chromatin alteration. As shown in Figures 1D–1F, the pattern of perinucleolar and perinuclear H3K27me3 (K27 trimethylated histone H3) staining was changed in a significant fraction of En1+/− TH+ neurons. The size distribution of the DAPI-dense regions of heterochromatin (Guenatri et al., 2004Guenatri M. Bailly D. Maison C. Almouzni G. Mouse centric and pericentric satellite repeats form distinct functional heterochromatin.J. Cell Biol. 2004; 166: 493-505Crossref PubMed Scopus (382) Google Scholar) also shows a reduction in the percentage of large spots in the En1+/− mouse (Figure 1G). Heterochromatin changes often are associated with changes in the expression of long interdispersed nuclear elements (LINEs) (Beisel and Paro, 2011Beisel C. Paro R. Silencing chromatin: comparing modes and mechanisms.Nat. Rev. Genet. 2011; 12: 123-135Crossref PubMed Scopus (286) Google Scholar, Guetg and Santoro, 2012Guetg C. Santoro R. Formation of nuclear heterochromatin: the nucleolar point of view.Epigenetics. 2012; 7: 811-814Crossref PubMed Scopus (63) Google Scholar); accordingly, LINE expression was increased in the SNpc of En1+/− mutants (Figure S1G). The induction of apoptosis genes (Figure 1A) was confirmed by the TH/activated caspase-3 co-staining in the SNpc of En1+/− mice, never seen in WT littermates (Figure S1H). An acute oxidative stress was applied to mDA neurons by injecting 6-OHDA (a superoxide-producing drug specifically captured by mDA neurons) directly into the SNpc. This induced, within 6 hr, the loss of 35% of the TH+ neurons and the formation of multiple abnormal γ-H2AX foci in about 26% of the remaining ones (Figures 2A and 2B ), only in the ipsilateral SNpc. Neuronal loss was paralleled by a reduction in TH protein and mRNA (En1 mRNA also) (Figures S2A and S2B) and the presence of numerous activated caspase-3-positive mDA neurons (Figure S2C). Injecting 6-OHDA into En1+/− mice led to a higher percentage of TH+ neurons with γ-H2AX foci in the 6-OHDA-injected side (51%) and to a parallel increase in the loss of TH+ neurons (60%) (Figures 2A and 2B). This demonstrates that endogenous En1 has a protective effect against oxidative stress. En1 is primarily a repressor gene (Tolkunova et al., 1998Tolkunova E.N. Fujioka M. Kobayashi M. Deka D. Jaynes J.B. Two distinct types of repression domain in engrailed: one interacts with the groucho corepressor and is preferentially active on integrated target genes.Mol. Cell. Biol. 1998; 18: 2804-2814Crossref PubMed Scopus (131) Google Scholar). Therefore, an activator form of Engrailed, composed of the En1/2 homeodomain (for target recognition) fused to four copies of the VP16 transcriptional activator domain of the herpes virus (EnHD-VP64) should have an anti-En1 activity. EnHD-VP64 infusion indeed leads to mDA cell death (Figures S3A and S3B) and to an increase in the number of γ-H2AX foci (Figure S3C). This confirms that Engrailed protects against oxidative stress, in part through transcriptional repression. Heterochromatin marks also were modified 6 hr after 6-OHDA injection in WT mice. The dense perinucleolar and perinuclear H3K27me3 staining in TH+ neurons of sham-injected mice was changed into diffuse nucleoplasmic staining in 6-OHDA-injected mice (Figure 2C). This change was quantified at the level of the nucleolus by measuring perinucleolar fluorescence intensity along one diameter (Figure 2D, left). Similarly, the ratio of H3K27me3 fluorescence intensities between the peripheral nuclear lamina and the nuclear stroma dropped from 1.4 to 1.0 (Figure 2D, right). Acute 6-OHDA also disrupted H3K9me3 (K9 trimethylated histone H3) and MeCP2 staining (Figures S2D and S2E), with the loss of neat lamin B2 staining (Figure S2F) and a change in the size distribution of DAPI-dense spots (Figure S2G). MeCP2 binds methylated CpGs and changes in its staining might reflect guanine oxidation (Skene et al., 2010Skene P.J. Illingworth R.S. Webb S. Kerr A.R. James K.D. Turner D.J. Andrews R. Bird A.P. Neuronal MeCP2 is expressed at near histone-octamer levels and globally alters the chromatin state.Mol. Cell. 2010; 37: 457-468Abstract Full Text Full Text PDF PubMed Scopus (502) Google Scholar). Nucleolar stress, suggested by perinucleolar γ-H2AX loss upon 6-OHDA (Figure 2A), was verified by the drop from 70% (sham) to 30% (6-OHDA) of mDA cells with dense nucleolin staining (Figures 2E and 2F). This change was accompanied by a strong upregulation of ribosomal pre-45S RNA (Figure 2G), signaling nucleolar damage (Guetg and Santoro, 2012Guetg C. Santoro R. Formation of nuclear heterochromatin: the nucleolar point of view.Epigenetics. 2012; 7: 811-814Crossref PubMed Scopus (63) Google Scholar, Larson et al., 2012Larson K. Yan S.J. Tsurumi A. Liu J. Zhou J. Gaur K. Guo D. Eickbush T.H. Li W.X. Heterochromatin formation promotes longevity and represses ribosomal RNA synthesis.PLoS Genet. 2012; 8: e1002473Crossref PubMed Scopus (199) Google Scholar). In comparison, nucleolin staining, normal in En1+/− TH+ neurons at 6 weeks, showed signs of disruption at 1 year (Figure 2H). However, the expression of genes involved in nucleolus organization (RNA-seq) and qRT-PCR analysis of pre-45S rRNA suggested a change in nucleolar physiology in 6- to 8-week-old En1+/− mice (Figures S3D and S3E). Nucleolar disruption links to the p53 pathways for senescence and apoptosis (Teng et al., 2013Teng T. Thomas G. Mercer C.A. Growth control and ribosomopathies.Curr. Opin. Genet. Dev. 2013; 23: 63-71Crossref PubMed Scopus (85) Google Scholar), including in mDA neurons (Rieker et al., 2011Rieker C. Engblom D. Kreiner G. Domanskyi A. Schober A. Stotz S. Neumann M. Yuan X. Grummt I. Schütz G. Parlato R. Nucleolar disruption in dopaminergic neurons leads to oxidative damage and parkinsonism through repression of mammalian target of rapamycin signaling.J. Neurosci. 2011; 31: 453-460Crossref PubMed Scopus (117) Google Scholar). The qRT-PCR on RNA from SNpc of sham- and 6-OHDA-injected mice showed that the level of p53 transcripts was increased by 50% upon 6-OHDA injection in parallel with that of p21, a p53 target (Sperka et al., 2012Sperka T. Wang J. Rudolph K.L. DNA damage checkpoints in stem cells, ageing and cancer.Nat. Rev. Mol. Cell Biol. 2012; 13: 579-590Crossref PubMed Scopus (313) Google Scholar; Figure S2H). Finally, in line with the regulation of retrotransposition through MeCP2 (Muotri et al., 2010Muotri A.R. Marchetto M.C. Coufal N.G. Oefner R. Yeo G. Nakashima K. Gage F.H. L1 retrotransposition in neurons is modulated by MeCP2.Nature. 2010; 468: 443-446Crossref PubMed Scopus (480) Google Scholar), LINE-1 mRNA levels were increased upon 6-OHDA injection (Figure S2I). To verify if Engrailed protects against oxidative stress, WT mice were unilaterally injected with 6-OHDA, and re-injected 30 min later with vehicle (sham) or En2. Analyses were done at 6 hr, 24 hr, and 7 days post-injections. The dramatic loss at 24 hr of TH+ cells in the SNpc of 6-OHDA/sham-injected mice was highly reduced in 6-OHDA/En2-injected mice (Figure 3A). Protection was still visible at 7 days with 40% and 20% of surviving neurons in En2-injected and sham mice, respectively (Figure 3B). En2 injection 24 hr (instead of 30 min) after 6-OHDA and analyzed 6 hr later showed no recovery, demonstrating that TH staining corresponds to true survival and not to TH re-expression (Figure S3F). As in other models (Casafont et al., 2011Casafont I. Palanca A. Lafarga V. Berciano M.T. Lafarga M. Effect of ionizing radiation in sensory ganglion neurons: organization and dynamics of nuclear compartments of DNA damage/repair and their relationship with transcription and cell cycle.Acta Neuropathol. 2011; 122: 481-493Crossref PubMed Scopus (32) Google Scholar, Li et al., 2013Li J. Hart R.P. Mallimo E.M. Swerdel M.R. Kusnecov A.W. Herrup K. EZH2-mediated H3K27 trimethylation mediates neurodegeneration in ataxia-telangiectasia.Nat. Neurosci. 2013; 16: 1745-1753Crossref PubMed Scopus (113) Google Scholar), 6-OHDA-induced apoptosis was paralleled by the expression of cell cycle markers (Figures S4A–S4D). Accordingly, cyclin A expression detected in En2-treated TH+ cells 6 hr post-injections had almost disappeared at 24 hr (Figure 3C). Neuronal rescue by En2 was associated with the reappearance of normal staining patterns for H3K27me3, nucleolin, and γ-H2AX (Figure 3D). The percentage of TH+ cells with WT perinucleolar H3K27me3 staining increased from 20% to 37% at 6 hr and reached 60% and 80% at 24 hr and 7 days, respectively. WT nucleolin pattern took longer to reappear as recovery was observed only at 24 hr with little changes between 24 hr and 1 week. Finally, the decrease in the number of γ-H2AX foci was slower with full recovery only at 7 days. Rescue correlated with a significant increase in TH mRNA expression and a decrease in that of pre-45S rRNA already 6 hr after En2 injection (Figure 3E). The expression of selected genes related to cell cycle and apoptosis (Lim and Kaldis, 2013Lim S. Kaldis P. Cdks, cyclins and CKIs: roles beyond cell cycle regulation.Development. 2013; 140: 3079-3093Crossref PubMed Scopus (1028) Google Scholar, Smith et al., 2003Smith P.D. Crocker S.J. Jackson-Lewis V. Jordan-Sciutto K.L. Hayley S. Mount M.P. O’Hare M.J. Callaghan S. Slack R.S. Przedborski S. et al.Cyclin-dependent kinase 5 is a mediator of dopaminergic neuron loss in a mouse model of Parkinson’s disease.Proc. Natl. Acad. Sci. USA. 2003; 100: 13650-13655Crossref PubMed Scopus (268) Google Scholar, Wang et al., 2009Wang W. Bu B. Xie M. Zhang M. Yu Z. Tao D. Neural cell cycle dysregulation and central nervous system diseases.Prog. Neurobiol. 2009; 89: 1-17Crossref PubMed Scopus (125) Google Scholar), upregulated in the En1+/− mouse or upon 6-OHDA injection, was repressed in the SNpc of 6-OHDA/En2 mice (Figure 3E). Analyses were done at 6 hr when sham- and En2-injected animals still have similar numbers of mDA neurons (Figure 3B). VTA neurons are less sensitive to the loss of one En1 allele. This might reflect the expression of Otx2, another transducing HP (Prochiantz and Di Nardo, 2015Prochiantz A. Di Nardo A.A. Homeoprotein signaling in the developing and adult nervous system.Neuron. 2015; 85: 911-925Abstract Full Text Full Text PDF PubMed Scopus (44) Google Scholar), specifically in the VTA (Di Salvio et al., 2010bDi Salvio M. Di Giovannantonio L.G. Omodei D. Acampora D. Simeone A. Otx2 expression is restricted to dopaminergic neurons of the ventral tegmental area in the adult brain.Int. J. Dev. Biol. 2010; 54: 939-945Crossref PubMed Scopus (66) Google Scholar) and its enhanced expression in the En1 mutant (Figure S1D). A protective effect of Otx2 was confirmed in the 6-OHDA model (Figure 3F). Conversely, Engrailed could act as a general midbrain survival factor. To address this point, embryonic day (E)14.5 midbrain neurons expressing Engrailed, but of which only 1%–2% were dopaminergic, were exposed to H202 with or without En2. Figure S4E illustrates that En2 decreased the number of DNA strand breaks induced by H2O2 in parallel with a decrease in the formation of comet tails that signal DNA damage. Protection by En2 requires its injection within 24 hr after 6-OHDA (Figure S3F). To identify genes in this early survival pathway, SNpc RNA from 6-OHDA/sham or 6-OHDA/En2 mice (6 hr) was sequenced and genes in the chromatin remodeling, DNA damage, apoptosis, and cell cycle pathways were analyzed according to Pathways Studio. Figures 4A and 4B rank by order of significance the genes with highest differences in read numbers, and Figure 4C provides qRT-PCR confirmation of enhanced expression for top representative genes in the four analyzed pathways. Genes in the apoptosis pathway were more represented than in the En1+/− mouse (Figure 1). This was not due to En2 injection per se (Figure S4F), suggesting that the rapid upregulation of anti-apoptotic pathways by En2 takes place specifically following the acute oxidative stress. Paralleling Gadd45b decreased expression in 8-week-old En1+/− mice, the addition of cycloheximide in the rescue experiments demonstrated that Gadd45b, as opposed to Pml for control, did not require the translation of an intervening protein and is, thus, a direct target of Engrailed (Figure 4D). The high induction of Gadd45b/g and NF-κB suggested a role for c-Jun N-terminal kinase (JNK) signaling, a pathway implied in several neurodegenerative diseases including PD (Coffey, 2014Coffey E.T. Nuclear and cytosolic JNK signalling in neurons.Nat. Rev. Neurosci. 2014; 15: 285-299Crossref PubMed Scopus (233) Google Scholar). Figures 4E and 4F confirm that the strong increase in p-JNK staining 6 hr after 6-OHDA was antagonized by En2 (Figure 4F). High levels of reactive oxygen species (ROS) are toxic, in particular at the DNA level (Vijg and Suh, 2013Vijg J. Suh Y. Genome instability and aging.Annu. Rev. Physiol. 2013; 75: 645-668Crossref PubMed Scopus (252) Google Scholar) where they directly induce DDR (O’Sullivan and Karlseder, 2012O’Sullivan R.J. Karlseder J. The great unravelling: chromatin as a modulator of the aging process.Trends Biochem. Sci. 2012; 37: 466-476Abstract Full Text Full Text PDF PubMed Scopus (82) Google Scholar). In consequence, neurons with high metabolic activity, such as SNpc mDA neurons, producing high amounts of ATP and ROS, are at risk for degeneration (Canugovi et al., 2013Canugovi C. Misiak M. Ferrarelli L.K. Croteau D.L. Bohr V.A. The role of DNA repair in brain related disease pathology.DNA Repair (Amst.). 2013; 12: 578-587Crossref PubMed Scopus (100) Google Scholar). Chromatin remodeling and DDR pathways are interconnected as DNA damage induces chromatin changes, themselves necessary to give access to the DNA repair machinery (Madabhushi et al., 2014Madabhushi R. Pan L. Tsai L.H. DNA damage and its links to neurodegeneration.Neuron. 2014; 83: 266-282Abstract Full Text Full Text PDF PubMed Scopus (384) Google Scholar, Soria et al., 2012Soria G. Polo S.E. Almouzni G. Prime, repair, restore: the active role of chromatin in the DNA damage response.Mol. Cell. 2012; 46: 722-734Abstract Full Text Full Text PDF PubMed Scopus (265) Google Scholar). This study places Engrailed genes as key regulators of DNA damage and chromatin changes that accompany chronic and acute forms of oxidative stress in mDA neurons. The En+/− chronic model of oxidative stress in which mDA neurons show progressive but faster death rate than in WT mice demonstrates that, similar to Otx2 dosage in the adult retina (Bernard et al., 2014Bernard C. Kim H.T. Torero Ibad R. Lee E.J. Simonutti M. Picaud S. Acampora D. Simeone A. Di Nardo A.A. Prochiantz A. et al.Graded Otx2 activities demonstrate dose-sensitive eye and retina phenotypes.Hum. Mol. Genet. 2014; 23: 1742-1753Crossref PubMed Scopus (27) Google Scholar), Engrailed dosage is important in the adult SNpc. Indeed, given that En1 and En2 are biochemically equivalent (Hanks et al., 1995Hanks M. Wurst W. Anson-Cartwright L. Auerbach A.B. Joyner A.L. Rescue of the En-1 mutant phenotype by replacement of En-1 with En-2.Science. 1995; 269: 679-682Crossref PubMed Scopus (359) Google Scholar), the loss of only one allele out of four is enough to accelerate cell death. In the context of aging and neurological diseases, this suggests that mDA neurons in the En1+/− mouse age faster and are more sensitive to 6-OHDA, a toxin used in animal models of PD. PD, even in its familial forms, declares itself rather late in life, underscoring a risk associated with age. Even if Engrailed is not a PD gene, its anti-aging properties might explain the association of EN1 polymorphisms and the risk to develop PD (Fuchs et al., 2009Fuchs J. Mueller J.C. Lichtner P. Schulte C. Munz M. Berg D. Wüllner U. Illig T. Sharma M. Gasser T. The transcription factor PITX3 is associated with sporadic Parkinson’s disease.Neurobiol. Aging. 2009; 30: 731-738Abstract Full Text Full Text PDF PubMed Scopus (86) Google Scholar, Haubenberger et al., 2011Haubenberger D. Reinthaler E. Mueller J.C. Pirker W. Katzenschlager R. Froehlich R. Bruecke T. Daniel G. Auff E. Zimprich A. Association of transcription factor polymorphisms PITX3 and EN1 with Parkinson’s disease.Neurobiol. Aging. 2011; 32: 302-307Abstract Full Text Full Text PDF PubMed Scopus (60) Google Scholar, Rissling et al., 2009Rissling I. Strauch K. Höft C. Oertel W.H. Möller J.C. Haplotype analysis of the engrailed-2 gene in young-onset Parkinson’s disease.Neurodegener. Dis. 2009; 6: 102-105Crossref P" @default.
• W2111034192 created "2016-06-24" @default.
• W2111034192 creator A5004015422 @default.
• W2111034192 creator A5005224351 @default.
• W2111034192 creator A5062243754 @default.
• W2111034192 creator A5065562137 @default.
• W2111034192 creator A5083790396 @default.
• W2111034192 date "2015-10-01" @default.
• W2111034192 modified "2023-10-18" @default.
• W2111034192 title "Engrailed Homeoprotein Protects Mesencephalic Dopaminergic Neurons from Oxidative Stress" @default.
• W2111034192 cites W1968170156 @default.
• W2111034192 cites W1974186613 @default.
• W2111034192 cites W1974987314 @default.
• W2111034192 cites W1979420039 @default.
• W2111034192 cites W1981149424 @default.
• W2111034192 cites W1984769882 @default.
• W2111034192 cites W1985139403 @default.
• W2111034192 cites W1990418954 @default.
• W2111034192 cites W1991327249 @default.
• W2111034192 cites W1993634104 @default.
• W2111034192 cites W1996683722 @default.
• W2111034192 cites W1999940145 @default.
• W2111034192 cites W2001936578 @default.
• W2111034192 cites W2014508690 @default.
• W2111034192 cites W2015734897 @default.
• W2111034192 cites W2018834244 @default.
• W2111034192 cites W2028597345 @default.
• W2111034192 cites W2031517458 @default.
• W2111034192 cites W2033700583 @default.
• W2111034192 cites W2034104365 @default.
• W2111034192 cites W2035488182 @default.
• W2111034192 cites W2042673772 @default.
• W2111034192 cites W2043422368 @default.
• W2111034192 cites W2047878192 @default.
• W2111034192 cites W2047919620 @default.
• W2111034192 cites W2055981668 @default.
• W2111034192 cites W2056066578 @default.
• W2111034192 cites W2067603001 @default.
• W2111034192 cites W2071061394 @default.
• W2111034192 cites W2073751853 @default.
• W2111034192 cites W2075758387 @default.
• W2111034192 cites W2080053685 @default.
• W2111034192 cites W2091442625 @default.
• W2111034192 cites W2105298120 @default.
• W2111034192 cites W2105733199 @default.
• W2111034192 cites W2114191977 @default.
• W2111034192 cites W2116326813 @default.
• W2111034192 cites W2117724384 @default.
• W2111034192 cites W2117743221 @default.
• W2111034192 cites W2120933894 @default.
• W2111034192 cites W2127631049 @default.
• W2111034192 cites W2128244475 @default.
• W2111034192 cites W2134020800 @default.
• W2111034192 cites W2141023520 @default.
• W2111034192 cites W2157101683 @default.
• W2111034192 cites W2160352834 @default.
• W2111034192 cites W2160740947 @default.
• W2111034192 cites W2170561922 @default.
• W2111034192 doi "https://doi.org/10.1016/j.celrep.2015.08.076" @default.
• W2111034192 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/5066840" @default.
• W2111034192 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/26411690" @default.
• W2111034192 hasPublicationYear "2015" @default.
• W2111034192 type Work @default.
• W2111034192 sameAs 2111034192 @default.
• W2111034192 citedByCount "43" @default.
• W2111034192 countsByYear W21110341922016 @default.
• W2111034192 countsByYear W21110341922017 @default.
• W2111034192 countsByYear W21110341922018 @default.
• W2111034192 countsByYear W21110341922019 @default.
• W2111034192 countsByYear W21110341922020 @default.
• W2111034192 countsByYear W21110341922021 @default.
• W2111034192 countsByYear W21110341922022 @default.
• W2111034192 countsByYear W21110341922023 @default.
• W2111034192 crossrefType "journal-article" @default.
• W2111034192 hasAuthorship W2111034192A5004015422 @default.
• W2111034192 hasAuthorship W2111034192A5005224351 @default.
• W2111034192 hasAuthorship W2111034192A5062243754 @default.
• W2111034192 hasAuthorship W2111034192A5065562137 @default.
• W2111034192 hasAuthorship W2111034192A5083790396 @default.
• W2111034192 hasBestOaLocation W21110341921 @default.
• W2111034192 hasConcept C137183658 @default.
• W2111034192 hasConcept C169760540 @default.
• W2111034192 hasConcept C185592680 @default.
• W2111034192 hasConcept C2776151105 @default.
• W2111034192 hasConcept C513476851 @default.
• W2111034192 hasConcept C529278444 @default.
• W2111034192 hasConcept C552161191 @default.
• W2111034192 hasConcept C55493867 @default.
• W2111034192 hasConcept C57600042 @default.
• W2111034192 hasConcept C86803240 @default.
• W2111034192 hasConcept C95444343 @default.
• W2111034192 hasConceptScore W2111034192C137183658 @default.
• W2111034192 hasConceptScore W2111034192C169760540 @default.
• W2111034192 hasConceptScore W2111034192C185592680 @default.
• W2111034192 hasConceptScore W2111034192C2776151105 @default.
• W2111034192 hasConceptScore W2111034192C513476851 @default.
• W2111034192 hasConceptScore W2111034192C529278444 @default.
• W2111034192 hasConceptScore W2111034192C552161191 @default.

• next