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Astrocyte-specific regulation of hMeCP2 expression in Drosophila.

Hess-Homeier DL, Fan CY, Gupta T, Chiang AS, Certel SJ - Biol Open (2014)

Bottom Line: Depending on the cell-type, hMeCP2 expression reduced sleep levels, altered daytime/nighttime sleep patterns, and generated sleep maintenance deficits.Levels of the full-length hMeCP2e2 and mutant RTT R106W protein decreased in astrocytes in a temporally and spatially regulated manner.In contrast, expression of the deletion Δ166 hMeCP2 protein was not altered in the entire astrocyte population. qPCR experiments revealed a reduction in full-length hMeCP2e2 transcript levels suggesting transgenic hMeCP2 expression is regulated at the transcriptional level.

View Article: PubMed Central - PubMed

Affiliation: Division of Biological Sciences, The University of Montana, Missoula, MT 59812, USA.

No MeSH data available.


Related in: MedlinePlus

hMeCP2 expression is developmentally regulated and does not alter glial differentiation.(A) At 1 day post-eclosion, hMeCP2 expression is detected by immunofluorescence in the majority of alrm-Gal4 expressing astrocytes. (B) At 5 days post-eclosion, hMeCP2 protein is reduced or not detectable in astrocytes located outside of the SOG region. (C) The transcription factor Repo is co-expressed with β-gal in optic lobe astrocytes (arrow) in alrm-Gal4/UAS-nucLacZ;alrm-Gal4 progeny. (D) hMeCP2-expressing astrocytes maintain glial differentiation as assayed by Repo expression in the optic lobe (arrow) and the SOG (arrowhead) in the alrm-Gal4/+;alrm-Gal4/UAS-hMeCP2FL adult brain. Scale bars represent 50 µm. (E,E′) Confocal image of the large number of astrocytes labeled by a cell-membrane GFP reporter in the brain of a control alrm-Gal4/+;alrm-Gal4/UAS-CD8:GFP adult. (E′) Optical sections of a hs-flp;alrm-Gal4/UAS->stop>CD8:GFP;alrm-Gal4/+ control brain at a high magnification showing a single GFP-expressing astrocyte. (F,F′) hMeCP2FL-expressing astrocytes labeled with the same cell-membrane GFP reporter in an adult alrm-Gal4/UAS-CD8:GFP;alrm-Gal4/UAS-hMeCP2FL brain five days post-eclosion. GFP-expressing astrocytes with or without detectable hMeCP2 (red nuclear expression) cover the entire brain as in control brains. (F′) Optical sections from a hs-flp;alrm-Gal4/UAS->stop>CD8:GFP;alrm-Gal4/UAS-hMeCP2FLbrain at high magnification showing a single GFP and hMeCP2FL -expressing astrocyte. The dense, fine processes observed in control astrocytes are present in astrocytes expressing hMeCP2FL. Scale bar represents 50 µm.
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f03: hMeCP2 expression is developmentally regulated and does not alter glial differentiation.(A) At 1 day post-eclosion, hMeCP2 expression is detected by immunofluorescence in the majority of alrm-Gal4 expressing astrocytes. (B) At 5 days post-eclosion, hMeCP2 protein is reduced or not detectable in astrocytes located outside of the SOG region. (C) The transcription factor Repo is co-expressed with β-gal in optic lobe astrocytes (arrow) in alrm-Gal4/UAS-nucLacZ;alrm-Gal4 progeny. (D) hMeCP2-expressing astrocytes maintain glial differentiation as assayed by Repo expression in the optic lobe (arrow) and the SOG (arrowhead) in the alrm-Gal4/+;alrm-Gal4/UAS-hMeCP2FL adult brain. Scale bars represent 50 µm. (E,E′) Confocal image of the large number of astrocytes labeled by a cell-membrane GFP reporter in the brain of a control alrm-Gal4/+;alrm-Gal4/UAS-CD8:GFP adult. (E′) Optical sections of a hs-flp;alrm-Gal4/UAS->stop>CD8:GFP;alrm-Gal4/+ control brain at a high magnification showing a single GFP-expressing astrocyte. (F,F′) hMeCP2FL-expressing astrocytes labeled with the same cell-membrane GFP reporter in an adult alrm-Gal4/UAS-CD8:GFP;alrm-Gal4/UAS-hMeCP2FL brain five days post-eclosion. GFP-expressing astrocytes with or without detectable hMeCP2 (red nuclear expression) cover the entire brain as in control brains. (F′) Optical sections from a hs-flp;alrm-Gal4/UAS->stop>CD8:GFP;alrm-Gal4/UAS-hMeCP2FLbrain at high magnification showing a single GFP and hMeCP2FL -expressing astrocyte. The dense, fine processes observed in control astrocytes are present in astrocytes expressing hMeCP2FL. Scale bar represents 50 µm.

Mentions: A developmental transition in hMeCP2 levels is also observed after larval stages. hMeCP2 expression is visible in all alrm-Gal4 astrocytes throughout the brain of newly-eclosed adults (Fig. 3A). The striking reduction in detectable hMeCP2 expression as described in Figs 1 and 2 is visualized by 5 days post-eclosion with expression of hMeCP2 maintained in SOG astrocytes of aged (25 day post-eclosion, Fig. 3B) adults. Results from these experiments suggest the molecular composition of astrocytes regionally, i.e. within the optic lobe and temporally, i.e. at mature stages (adult and larval) include a gene product that regulates hMeCP2 at the transcriptional or translational level.


Astrocyte-specific regulation of hMeCP2 expression in Drosophila.

Hess-Homeier DL, Fan CY, Gupta T, Chiang AS, Certel SJ - Biol Open (2014)

hMeCP2 expression is developmentally regulated and does not alter glial differentiation.(A) At 1 day post-eclosion, hMeCP2 expression is detected by immunofluorescence in the majority of alrm-Gal4 expressing astrocytes. (B) At 5 days post-eclosion, hMeCP2 protein is reduced or not detectable in astrocytes located outside of the SOG region. (C) The transcription factor Repo is co-expressed with β-gal in optic lobe astrocytes (arrow) in alrm-Gal4/UAS-nucLacZ;alrm-Gal4 progeny. (D) hMeCP2-expressing astrocytes maintain glial differentiation as assayed by Repo expression in the optic lobe (arrow) and the SOG (arrowhead) in the alrm-Gal4/+;alrm-Gal4/UAS-hMeCP2FL adult brain. Scale bars represent 50 µm. (E,E′) Confocal image of the large number of astrocytes labeled by a cell-membrane GFP reporter in the brain of a control alrm-Gal4/+;alrm-Gal4/UAS-CD8:GFP adult. (E′) Optical sections of a hs-flp;alrm-Gal4/UAS->stop>CD8:GFP;alrm-Gal4/+ control brain at a high magnification showing a single GFP-expressing astrocyte. (F,F′) hMeCP2FL-expressing astrocytes labeled with the same cell-membrane GFP reporter in an adult alrm-Gal4/UAS-CD8:GFP;alrm-Gal4/UAS-hMeCP2FL brain five days post-eclosion. GFP-expressing astrocytes with or without detectable hMeCP2 (red nuclear expression) cover the entire brain as in control brains. (F′) Optical sections from a hs-flp;alrm-Gal4/UAS->stop>CD8:GFP;alrm-Gal4/UAS-hMeCP2FLbrain at high magnification showing a single GFP and hMeCP2FL -expressing astrocyte. The dense, fine processes observed in control astrocytes are present in astrocytes expressing hMeCP2FL. Scale bar represents 50 µm.
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Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4232758&req=5

f03: hMeCP2 expression is developmentally regulated and does not alter glial differentiation.(A) At 1 day post-eclosion, hMeCP2 expression is detected by immunofluorescence in the majority of alrm-Gal4 expressing astrocytes. (B) At 5 days post-eclosion, hMeCP2 protein is reduced or not detectable in astrocytes located outside of the SOG region. (C) The transcription factor Repo is co-expressed with β-gal in optic lobe astrocytes (arrow) in alrm-Gal4/UAS-nucLacZ;alrm-Gal4 progeny. (D) hMeCP2-expressing astrocytes maintain glial differentiation as assayed by Repo expression in the optic lobe (arrow) and the SOG (arrowhead) in the alrm-Gal4/+;alrm-Gal4/UAS-hMeCP2FL adult brain. Scale bars represent 50 µm. (E,E′) Confocal image of the large number of astrocytes labeled by a cell-membrane GFP reporter in the brain of a control alrm-Gal4/+;alrm-Gal4/UAS-CD8:GFP adult. (E′) Optical sections of a hs-flp;alrm-Gal4/UAS->stop>CD8:GFP;alrm-Gal4/+ control brain at a high magnification showing a single GFP-expressing astrocyte. (F,F′) hMeCP2FL-expressing astrocytes labeled with the same cell-membrane GFP reporter in an adult alrm-Gal4/UAS-CD8:GFP;alrm-Gal4/UAS-hMeCP2FL brain five days post-eclosion. GFP-expressing astrocytes with or without detectable hMeCP2 (red nuclear expression) cover the entire brain as in control brains. (F′) Optical sections from a hs-flp;alrm-Gal4/UAS->stop>CD8:GFP;alrm-Gal4/UAS-hMeCP2FLbrain at high magnification showing a single GFP and hMeCP2FL -expressing astrocyte. The dense, fine processes observed in control astrocytes are present in astrocytes expressing hMeCP2FL. Scale bar represents 50 µm.
Mentions: A developmental transition in hMeCP2 levels is also observed after larval stages. hMeCP2 expression is visible in all alrm-Gal4 astrocytes throughout the brain of newly-eclosed adults (Fig. 3A). The striking reduction in detectable hMeCP2 expression as described in Figs 1 and 2 is visualized by 5 days post-eclosion with expression of hMeCP2 maintained in SOG astrocytes of aged (25 day post-eclosion, Fig. 3B) adults. Results from these experiments suggest the molecular composition of astrocytes regionally, i.e. within the optic lobe and temporally, i.e. at mature stages (adult and larval) include a gene product that regulates hMeCP2 at the transcriptional or translational level.

Bottom Line: Depending on the cell-type, hMeCP2 expression reduced sleep levels, altered daytime/nighttime sleep patterns, and generated sleep maintenance deficits.Levels of the full-length hMeCP2e2 and mutant RTT R106W protein decreased in astrocytes in a temporally and spatially regulated manner.In contrast, expression of the deletion Δ166 hMeCP2 protein was not altered in the entire astrocyte population. qPCR experiments revealed a reduction in full-length hMeCP2e2 transcript levels suggesting transgenic hMeCP2 expression is regulated at the transcriptional level.

View Article: PubMed Central - PubMed

Affiliation: Division of Biological Sciences, The University of Montana, Missoula, MT 59812, USA.

No MeSH data available.


Related in: MedlinePlus