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Comparison of larval and adult Drosophila astrocytes reveals stage-specific gene expression profiles.

Huang Y, Ng FS, Jackson FR - G3 (Bethesda) (2015)

Bottom Line: These include genes important for metabolism and energy production, translation, chromatin modification, protein glycosylation, neuropeptide signaling, immune responses, vesicle-mediated trafficking or secretion, and the regulation of behavior.Among these functional classes, the expression of genes important for chromatin modification and vesicle-mediated trafficking or secretion is overrepresented in adult astrocytes based on Gene Ontology analysis.Certain genes with selective adult enrichment may mediate functions specific to this stage or may be important for the differentiation or maintenance of adult astrocytes, with the latter perhaps contributing to population heterogeneity.

View Article: PubMed Central - PubMed

Affiliation: Department of Neuroscience, Sackler School of Graduate Biomedical Sciences, Tufts University School of Medicine, Boston, Massachusetts 02111.

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Expression of EGFP-L10a in larval nervous system astrocytes. (A) The alrm-Gal4 driver was used to limit expression to astrocytes. (B) Low magnification view of the entire larval brain (Br) and ventral nerve cord (VNC) showing alrm-driven expression of EGFP-L10a in both regions. (C1 and C2) View of EGFP-L10a expression in brain astrocytes illustrating cytoplasmic localization of the protein. (D1 and D2) EGFP-L10a expression in the cytoplasm of VNC astrocytes. Green, EGFP-L10a; red, Repo (a glial nuclear marker); size bars = 50 μm in B, C1, and D1, and 10 μm in C2 and D2. The image in (B) represents a Z stack of optical sections representing the whole nervous system. Other images of this figure are 1.5-μm optical sections.
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fig1: Expression of EGFP-L10a in larval nervous system astrocytes. (A) The alrm-Gal4 driver was used to limit expression to astrocytes. (B) Low magnification view of the entire larval brain (Br) and ventral nerve cord (VNC) showing alrm-driven expression of EGFP-L10a in both regions. (C1 and C2) View of EGFP-L10a expression in brain astrocytes illustrating cytoplasmic localization of the protein. (D1 and D2) EGFP-L10a expression in the cytoplasm of VNC astrocytes. Green, EGFP-L10a; red, Repo (a glial nuclear marker); size bars = 50 μm in B, C1, and D1, and 10 μm in C2 and D2. The image in (B) represents a Z stack of optical sections representing the whole nervous system. Other images of this figure are 1.5-μm optical sections.

Mentions: We performed larval TRAP profiling using a previously described astrocyte Gal4 driver strain (alrm-Gal4) (Doherty et al. 2009). The alrm-Gal4 driver was combined with a Drosophila transgene encoding a tagged large ribosomal subunit (UAS-EGFP-L10a) (Huang et al. 2013) to generate flies specifically expressing EGFP-L10a in astrocytes of the larval nervous system (brain and ventral nerve cord; Figure 1A). As shown in Figure 1, EGFP-L10a could be detected in glial cells of the larval brain lobes (Figure 1, B and C) and ventral nerve cord (Figure 1, B and D). In both locations, EGFP-L10a was observed to be cytoplasmic, as determined by straining with the Repo nuclear marker (Figure 1, C and D). Expression of alrm-Gal4-driven EGFP-L10a throughout development did not grossly affect astrocyte cell morphology (Figure 1 and data not shown), nor did it affect viability or activity level or cause circadian arrhythmicity for adult animals (Supporting Information, Figure S1).


Comparison of larval and adult Drosophila astrocytes reveals stage-specific gene expression profiles.

Huang Y, Ng FS, Jackson FR - G3 (Bethesda) (2015)

Expression of EGFP-L10a in larval nervous system astrocytes. (A) The alrm-Gal4 driver was used to limit expression to astrocytes. (B) Low magnification view of the entire larval brain (Br) and ventral nerve cord (VNC) showing alrm-driven expression of EGFP-L10a in both regions. (C1 and C2) View of EGFP-L10a expression in brain astrocytes illustrating cytoplasmic localization of the protein. (D1 and D2) EGFP-L10a expression in the cytoplasm of VNC astrocytes. Green, EGFP-L10a; red, Repo (a glial nuclear marker); size bars = 50 μm in B, C1, and D1, and 10 μm in C2 and D2. The image in (B) represents a Z stack of optical sections representing the whole nervous system. Other images of this figure are 1.5-μm optical sections.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig1: Expression of EGFP-L10a in larval nervous system astrocytes. (A) The alrm-Gal4 driver was used to limit expression to astrocytes. (B) Low magnification view of the entire larval brain (Br) and ventral nerve cord (VNC) showing alrm-driven expression of EGFP-L10a in both regions. (C1 and C2) View of EGFP-L10a expression in brain astrocytes illustrating cytoplasmic localization of the protein. (D1 and D2) EGFP-L10a expression in the cytoplasm of VNC astrocytes. Green, EGFP-L10a; red, Repo (a glial nuclear marker); size bars = 50 μm in B, C1, and D1, and 10 μm in C2 and D2. The image in (B) represents a Z stack of optical sections representing the whole nervous system. Other images of this figure are 1.5-μm optical sections.
Mentions: We performed larval TRAP profiling using a previously described astrocyte Gal4 driver strain (alrm-Gal4) (Doherty et al. 2009). The alrm-Gal4 driver was combined with a Drosophila transgene encoding a tagged large ribosomal subunit (UAS-EGFP-L10a) (Huang et al. 2013) to generate flies specifically expressing EGFP-L10a in astrocytes of the larval nervous system (brain and ventral nerve cord; Figure 1A). As shown in Figure 1, EGFP-L10a could be detected in glial cells of the larval brain lobes (Figure 1, B and C) and ventral nerve cord (Figure 1, B and D). In both locations, EGFP-L10a was observed to be cytoplasmic, as determined by straining with the Repo nuclear marker (Figure 1, C and D). Expression of alrm-Gal4-driven EGFP-L10a throughout development did not grossly affect astrocyte cell morphology (Figure 1 and data not shown), nor did it affect viability or activity level or cause circadian arrhythmicity for adult animals (Supporting Information, Figure S1).

Bottom Line: These include genes important for metabolism and energy production, translation, chromatin modification, protein glycosylation, neuropeptide signaling, immune responses, vesicle-mediated trafficking or secretion, and the regulation of behavior.Among these functional classes, the expression of genes important for chromatin modification and vesicle-mediated trafficking or secretion is overrepresented in adult astrocytes based on Gene Ontology analysis.Certain genes with selective adult enrichment may mediate functions specific to this stage or may be important for the differentiation or maintenance of adult astrocytes, with the latter perhaps contributing to population heterogeneity.

View Article: PubMed Central - PubMed

Affiliation: Department of Neuroscience, Sackler School of Graduate Biomedical Sciences, Tufts University School of Medicine, Boston, Massachusetts 02111.

Show MeSH
Related in: MedlinePlus