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Figure 5 | Neural Development

Figure 5

From: Dynamics of glutamatergic signaling in the mushroom body of young adult Drosophila

Figure 5

Dynamics of glutamine synthetase expression in mushroom body glial cells after adult eclosion. (A) A mouse monoclonal antibody raised against sheep glutamine synthetase (GS) recognizes a single band in a western blot of Drosophila head and brain. The apparent size of the protein (approximately 42 kDa) closely corresponds to the predicted size of Drosophila GS2 (41 kDa). (B) Localization of GS immunoreactivity in glial cells that express the Glu transporter dEAAT1 in 10-day-old adult fly MBs. Triple staining of brains from dEAAT1-GAL4; UAS-dEAAT1::GFP Drosophila with anti-GS (magenta), anti-GFP (green) and anti-DC0 (blue, for MB staining). (B1-B3) GS co-localizes with dEAAT1-positive glial cell bodies and processes in the spur (sp) region of the MB neuropil (merged magenta and green produces a white color). (C) Localization of GS in the MB with respect to glutamatergic synapses. Triple staining of wild-type adult brains with anti-DVGluT (green), anti-GS (magenta), and anti-GFP (blue) in agarose sections. (C1) GS is distributed all over the lobes (arrowheads) except the core neurons. (C2) DVGluT-positive glutamatergic synapses are present on the α lobe (arrow). (C3) GS immunoreactivity lies in close vicinity to glutamatergic synapses (arrow). In the β lobe, GS is present (arrowheads) but DVGluT is absent. (D-I) Sections of Drosophila brains stained with anti-GS (magenta) at different adult ages showing the MB pedunculus (D, F, H) and the medial lobe region (E, G, I). MB α/β KCs are positive for GFP (green) in c739-GAL4; UAS-mCD8::GFP flies (D-G) or DC0 (green) in wild-type flies (H-I). In the pedunculus, GS is found in glial cells that surrounds the α/β axons either within 1 hour after adult eclosion (D), 24 hours later (F) or at 10 days old (adult) (H). (E) Within 1 hour after eclosion, GS-immunoreactive processes enwrap the MB medial β and γ lobes. GS is also detected in many profiles inside the β and γ lobes, except in the regions corresponding to the core neurons that express neither GFP nor GS. (G) Twenty-four hours after eclosion, GS-positive glial processes distribute evenly in the whole β lobe area, including the β core, which at this time expresses GFP. (I) Ten days after eclosion, GS is expressed at a high level in glial cells that surround the β lobes. Scale bars: 20 μm.

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