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. 2014 Jan 1;28(1):20–33. doi: 10.1101/gad.229518.113

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© 2014 Tasdemir-Yilmaz and Freeman; Published by Cold Spring Harbor Laboratory Press

This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 3.0 Unported), as described at http://creativecommons.org/licenses/by-nc/3.0/.

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Figure 3. — Astrocytes engulf and clear synaptic material from the neuropil. (A) In A and D, confocal images are single z-confocal slices. Astrocytes were labeled with GFP (alrm-Gal4, UAS-mCD8∷GFP/+; green), glial nuclei were labeled with anti-Repo (blue), and, in A and D, active zones were labeled with nc82 (antibody for Brp; red). Time points are as follows: L3 (third instar larva) and APF (hours APF). (Top inset) High-magnification view of the boxed region. (Middle and bottom insets) Other examples of vacuoles. Bars: 20 μm; insets, 1 μm. (B) The graph shows fluorescence intensity per pixel in each channel for the inset in A of the astrocyte membrane and nc82⁺ puncta along a line drawn through a vacuole. (C) TEM image from a cross-section of the abdominal VNC of an alrm>EcR^DN animal. An astrocyte (outlined in yellow) at the edge of the neuropil. Time point is as indicated (APF). Genotype used was as follows: alrm>EcR^DN (alrm-Gal4,UAS-mCD8∷GFP/UAS-EcR^DN). Bars, 2 μm. (D) Time points are as indicated (APF). Genotypes used were as follows: control (alrm-Gal4/+) and alrm>EcR^DN (alrm-Gal4/UAS-EcR^DN). Bars, 20 μm. (E,F) Quantification of pixel intensity of nc82⁺ puncta at L3 and 18 h APF in SOG, thorax, and abdominal regions of the VNC. (E) Genotypes used were as follows: EcR^DN/+ (UAS-EcR^DN/+) and alrm>EcR^DN (alrm-Gal4/UAS-EcR^DN). Error bars represent ±SEM. N-values are as follows: alrm-Gal4/+, N ≥ 20; UAS-EcR^DN/+, N ≥ 16; and alrm-Gal4/UAS-EcR^DN, N ≥ 20 (N denotes number of measurements). (*) P < 0.05; (**) P < 0.01; (****) P < 0.0001. (F) Genotypes used were as follows: shi^ts/+ (UAS-shi^ts/+) and alrm>shi^ts (alrm-Gal4/UAS-shi^ts). Flies were raised at 18°C and then shifted to the restrictive temperature (30°C) at 0 h APF for 18 h. Error bars represent ±SEM; N ≥ 12; (****) P < 0.0001.