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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

The hMeCP2Δ166 allele is expressed in all astrocytes.(A,B) hMeCP2Δ166 expression is detected in alrm-Gal4 expressing astrocytes throughout the brain at 5 (A) or 15 (B) days post-eclosion (alrm-Gal4/UAS- hMeCP2Δ166;alrm-Gal4/+). (C) Schematic representation of the hMeCP2Δ166 allele containing a deletion of the N-terminus including the MBD. hMeCP2Δ166 expression is detected by immunofluorescence with the rabbit hMeCP2 antibody (green, Cell Signaling). (D,D′) Confocal images of two UAS-dsRed;EAAT1-Gal4/UAS-hMeCP2FL adult brains demonstrating hMeCP2FL protein is largely absent or reduced in the Eaat1 glial population. Co-expression of dsRed and hMeCP2FL is maintained in a number of glia in the optic lobe (arrow) or around the antennal lobe. Scale bars represent 50 µm.
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f04: The hMeCP2Δ166 allele is expressed in all astrocytes.(A,B) hMeCP2Δ166 expression is detected in alrm-Gal4 expressing astrocytes throughout the brain at 5 (A) or 15 (B) days post-eclosion (alrm-Gal4/UAS- hMeCP2Δ166;alrm-Gal4/+). (C) Schematic representation of the hMeCP2Δ166 allele containing a deletion of the N-terminus including the MBD. hMeCP2Δ166 expression is detected by immunofluorescence with the rabbit hMeCP2 antibody (green, Cell Signaling). (D,D′) Confocal images of two UAS-dsRed;EAAT1-Gal4/UAS-hMeCP2FL adult brains demonstrating hMeCP2FL protein is largely absent or reduced in the Eaat1 glial population. Co-expression of dsRed and hMeCP2FL is maintained in a number of glia in the optic lobe (arrow) or around the antennal lobe. Scale bars represent 50 µm.

Mentions: Using a third independent transgenic line, we expressed hMeCP2Δ166, which lacks the N-terminus including the MBD (Fig. 4C) (Cukier et al., 2008), via the alrm-Gal4 driver. In contrast to the region-specific reduction in hMeCP2FL or hMeCP2R106W expression, the hMeCP2Δ166 protein is detected in the entire alrm-Gal4 astrocyte population and is maintained in the aging brain (Fig. 4A,B). This result indicates that transgenic expression of hMeCP2 in all alrm-Gal4 astrocytes is possible and suggests that the N-terminus contains a site that may be targeted by a cell-specific endogenous factor to transcriptionally or translationally regulate full-length hMeCP2 expression. Although astrocytes are often classified as a population of cells, glial subsets can display diverse molecular and functional profiles (Matthias et al., 2003; Nimmerjahn, 2009; Regan et al., 2007).


Astrocyte-specific regulation of hMeCP2 expression in Drosophila.

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

The hMeCP2Δ166 allele is expressed in all astrocytes.(A,B) hMeCP2Δ166 expression is detected in alrm-Gal4 expressing astrocytes throughout the brain at 5 (A) or 15 (B) days post-eclosion (alrm-Gal4/UAS- hMeCP2Δ166;alrm-Gal4/+). (C) Schematic representation of the hMeCP2Δ166 allele containing a deletion of the N-terminus including the MBD. hMeCP2Δ166 expression is detected by immunofluorescence with the rabbit hMeCP2 antibody (green, Cell Signaling). (D,D′) Confocal images of two UAS-dsRed;EAAT1-Gal4/UAS-hMeCP2FL adult brains demonstrating hMeCP2FL protein is largely absent or reduced in the Eaat1 glial population. Co-expression of dsRed and hMeCP2FL is maintained in a number of glia in the optic lobe (arrow) or around the antennal lobe. Scale bars represent 50 µm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f04: The hMeCP2Δ166 allele is expressed in all astrocytes.(A,B) hMeCP2Δ166 expression is detected in alrm-Gal4 expressing astrocytes throughout the brain at 5 (A) or 15 (B) days post-eclosion (alrm-Gal4/UAS- hMeCP2Δ166;alrm-Gal4/+). (C) Schematic representation of the hMeCP2Δ166 allele containing a deletion of the N-terminus including the MBD. hMeCP2Δ166 expression is detected by immunofluorescence with the rabbit hMeCP2 antibody (green, Cell Signaling). (D,D′) Confocal images of two UAS-dsRed;EAAT1-Gal4/UAS-hMeCP2FL adult brains demonstrating hMeCP2FL protein is largely absent or reduced in the Eaat1 glial population. Co-expression of dsRed and hMeCP2FL is maintained in a number of glia in the optic lobe (arrow) or around the antennal lobe. Scale bars represent 50 µm.
Mentions: Using a third independent transgenic line, we expressed hMeCP2Δ166, which lacks the N-terminus including the MBD (Fig. 4C) (Cukier et al., 2008), via the alrm-Gal4 driver. In contrast to the region-specific reduction in hMeCP2FL or hMeCP2R106W expression, the hMeCP2Δ166 protein is detected in the entire alrm-Gal4 astrocyte population and is maintained in the aging brain (Fig. 4A,B). This result indicates that transgenic expression of hMeCP2 in all alrm-Gal4 astrocytes is possible and suggests that the N-terminus contains a site that may be targeted by a cell-specific endogenous factor to transcriptionally or translationally regulate full-length hMeCP2 expression. Although astrocytes are often classified as a population of cells, glial subsets can display diverse molecular and functional profiles (Matthias et al., 2003; Nimmerjahn, 2009; Regan et al., 2007).

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