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Rapid clearance of epigenetic protein reporters from wound edge cells in Drosophila larvae does not depend on the JNK or PDGFR/VEGFR signaling pathways.

Anderson AE, Galko MJ - Regeneration (Oxf) (2014)

Bottom Line: Three downregulated proteins, Osa, Kismet, and Spt6, are generally associated with active chromatin, while four others, Sin3A, Sap130, Mi-2, and Mip120, are associated with repressed chromatin.In all cases reporter down regulation was independent of the Jun N-terminal Kinase and Pvr pathways, suggesting that novel signals control reporter clearance.Taken together, our results suggest that clearance of chromatin modifying factors may enable wound edge cells to rapidly and comprehensively change their transcriptional state following tissue damage.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, The University of Texas MD Anderson Cancer Center, Unit 1000, 1515 Holcombe Boulevard, Houston, TX 77030, USA.

ABSTRACT
The drastic cellular changes required for epidermal cells to dedifferentiate and become motile during wound closure are accompanied by changes in gene transcription, suggesting corresponding alterations in chromatin. However, the epigenetic changes that underlie wound-induced transcriptional programs remain poorly understood partly because a comprehensive study of epigenetic factor expression during wound healing has not been practical. To determine which chromatin modifying factors might contribute to wound healing, we screened publicly available fluorescently-tagged reporter lines in Drosophila for altered expression at the wound periphery during healing. Thirteen reporters tagging seven different proteins showed strongly diminished expression at the wound edge. Three downregulated proteins, Osa, Kismet, and Spt6, are generally associated with active chromatin, while four others, Sin3A, Sap130, Mi-2, and Mip120, are associated with repressed chromatin. In all cases reporter down regulation was independent of the Jun N-terminal Kinase and Pvr pathways, suggesting that novel signals control reporter clearance. Taken together, our results suggest that clearance of chromatin modifying factors may enable wound edge cells to rapidly and comprehensively change their transcriptional state following tissue damage.

No MeSH data available.


Related in: MedlinePlus

Two distinct temporal patterns of reporter regulation during wound healing. (A)−(H) Dissected epidermal whole mounts of larvae heterozygous for e22c‐Gal4, UAS‐DsRed2‐Nuc and the indicated reporter transgene were immunostained for Fasciclin III at the indicated times post‐wounding. Two examples are shown. In pattern 1 (A−D′, Sin3AR2 reporter), expression is lost from wound edge cells within minutes of wounding. In pattern 2 (E−H′, OsaR1 reporter), expression is lost between wounding and 4 h later. Scale bar 200 μm. (I) Graphical representation of reporter downregulation following wounding. Arrows indicate epidermal nuclei that have absent or reduced reporter expression. Yellow, epidermal nucleus expressing both DsRed and reporter; red, epidermal nucleus.
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reg212-fig-0003: Two distinct temporal patterns of reporter regulation during wound healing. (A)−(H) Dissected epidermal whole mounts of larvae heterozygous for e22c‐Gal4, UAS‐DsRed2‐Nuc and the indicated reporter transgene were immunostained for Fasciclin III at the indicated times post‐wounding. Two examples are shown. In pattern 1 (A−D′, Sin3AR2 reporter), expression is lost from wound edge cells within minutes of wounding. In pattern 2 (E−H′, OsaR1 reporter), expression is lost between wounding and 4 h later. Scale bar 200 μm. (I) Graphical representation of reporter downregulation following wounding. Arrows indicate epidermal nuclei that have absent or reduced reporter expression. Yellow, epidermal nucleus expressing both DsRed and reporter; red, epidermal nucleus.

Mentions: Next, we asked when the expression of each downregulated reporter first begins to diminish, and whether their expression is reestablished concomitant with wound closure. We therefore examined epidermal expression of each of the 13 reporters at 0, 8, and 12 h after wounding. We observed two types of expression pattern (Fig. 3I). Strikingly, the Sap130 and Sin3A reporters (type I) were cleared from wound edge cells within minutes of wounding (Figs. 2A−A′ and S3A−A′). Some nuclei that lack reporter expression can still be observed near the presumptive center of the closed wound 12 h later (arrows in Figs. 3D−D′ and S4D−D′). By contrast, reporters for Mip120, Kismet, Osa, Spt6, and Mi‐2 (type II) showed delayed clearance from wound edge cells, with the reporters present immediately post‐wounding (Figs. 3E−E′ and S4E−E′,I−I′,M−M′,Q−Q′) but absent at 4 h post‐wounding (Fig. 1A−D′′′,G−G′′′). However, as with type I reporters, weak or absent reporter staining was also observed at 12 h after wounding in most of these lines (Figs. 3H−H′ and S4H−H′,L−L′,P−P′,T−T′).


Rapid clearance of epigenetic protein reporters from wound edge cells in Drosophila larvae does not depend on the JNK or PDGFR/VEGFR signaling pathways.

Anderson AE, Galko MJ - Regeneration (Oxf) (2014)

Two distinct temporal patterns of reporter regulation during wound healing. (A)−(H) Dissected epidermal whole mounts of larvae heterozygous for e22c‐Gal4, UAS‐DsRed2‐Nuc and the indicated reporter transgene were immunostained for Fasciclin III at the indicated times post‐wounding. Two examples are shown. In pattern 1 (A−D′, Sin3AR2 reporter), expression is lost from wound edge cells within minutes of wounding. In pattern 2 (E−H′, OsaR1 reporter), expression is lost between wounding and 4 h later. Scale bar 200 μm. (I) Graphical representation of reporter downregulation following wounding. Arrows indicate epidermal nuclei that have absent or reduced reporter expression. Yellow, epidermal nucleus expressing both DsRed and reporter; red, epidermal nucleus.
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reg212-fig-0003: Two distinct temporal patterns of reporter regulation during wound healing. (A)−(H) Dissected epidermal whole mounts of larvae heterozygous for e22c‐Gal4, UAS‐DsRed2‐Nuc and the indicated reporter transgene were immunostained for Fasciclin III at the indicated times post‐wounding. Two examples are shown. In pattern 1 (A−D′, Sin3AR2 reporter), expression is lost from wound edge cells within minutes of wounding. In pattern 2 (E−H′, OsaR1 reporter), expression is lost between wounding and 4 h later. Scale bar 200 μm. (I) Graphical representation of reporter downregulation following wounding. Arrows indicate epidermal nuclei that have absent or reduced reporter expression. Yellow, epidermal nucleus expressing both DsRed and reporter; red, epidermal nucleus.
Mentions: Next, we asked when the expression of each downregulated reporter first begins to diminish, and whether their expression is reestablished concomitant with wound closure. We therefore examined epidermal expression of each of the 13 reporters at 0, 8, and 12 h after wounding. We observed two types of expression pattern (Fig. 3I). Strikingly, the Sap130 and Sin3A reporters (type I) were cleared from wound edge cells within minutes of wounding (Figs. 2A−A′ and S3A−A′). Some nuclei that lack reporter expression can still be observed near the presumptive center of the closed wound 12 h later (arrows in Figs. 3D−D′ and S4D−D′). By contrast, reporters for Mip120, Kismet, Osa, Spt6, and Mi‐2 (type II) showed delayed clearance from wound edge cells, with the reporters present immediately post‐wounding (Figs. 3E−E′ and S4E−E′,I−I′,M−M′,Q−Q′) but absent at 4 h post‐wounding (Fig. 1A−D′′′,G−G′′′). However, as with type I reporters, weak or absent reporter staining was also observed at 12 h after wounding in most of these lines (Figs. 3H−H′ and S4H−H′,L−L′,P−P′,T−T′).

Bottom Line: Three downregulated proteins, Osa, Kismet, and Spt6, are generally associated with active chromatin, while four others, Sin3A, Sap130, Mi-2, and Mip120, are associated with repressed chromatin.In all cases reporter down regulation was independent of the Jun N-terminal Kinase and Pvr pathways, suggesting that novel signals control reporter clearance.Taken together, our results suggest that clearance of chromatin modifying factors may enable wound edge cells to rapidly and comprehensively change their transcriptional state following tissue damage.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, The University of Texas MD Anderson Cancer Center, Unit 1000, 1515 Holcombe Boulevard, Houston, TX 77030, USA.

ABSTRACT
The drastic cellular changes required for epidermal cells to dedifferentiate and become motile during wound closure are accompanied by changes in gene transcription, suggesting corresponding alterations in chromatin. However, the epigenetic changes that underlie wound-induced transcriptional programs remain poorly understood partly because a comprehensive study of epigenetic factor expression during wound healing has not been practical. To determine which chromatin modifying factors might contribute to wound healing, we screened publicly available fluorescently-tagged reporter lines in Drosophila for altered expression at the wound periphery during healing. Thirteen reporters tagging seven different proteins showed strongly diminished expression at the wound edge. Three downregulated proteins, Osa, Kismet, and Spt6, are generally associated with active chromatin, while four others, Sin3A, Sap130, Mi-2, and Mip120, are associated with repressed chromatin. In all cases reporter down regulation was independent of the Jun N-terminal Kinase and Pvr pathways, suggesting that novel signals control reporter clearance. Taken together, our results suggest that clearance of chromatin modifying factors may enable wound edge cells to rapidly and comprehensively change their transcriptional state following tissue damage.

No MeSH data available.


Related in: MedlinePlus