Journal: PLoS Genetics

Article Title: C-terminal Src Kinase Gates Homeostatic Synaptic Plasticity and Regulates Fasciclin II Expression at the Drosophila Neuromuscular Junction

doi: 10.1371/journal.pgen.1005886

Figure Lengend Snippet: (A-E) Representative images of FasII immunostaining at NMJs that are (A) wild type, (B) Csk c04256 /Csk j1D8 , (C) expressing Csk-RNAi in muscle and neurons, (D) expressing Csk-RNAi in only muscle, and (E) expressing Csk-RNAi in only neurons. (F) Average values for synaptic FasII fluorescence intensity normalized to synapse area and normalized to wild type. (G) Western blot of FasII (DSHB 1D4 antibody) in protein extracts from whole third instar larvae. (H, I) Relative quantification of (H) total FasII intensity and (I) the intensity of the lowest molecular weight FasII band (the ‘third band’). Quantification in H and I is shown as a fold change relative to wild type. Values on/above bars indicate the number of biological replicates for each genotype. * p < 0.05, ** p < 0.01, *** p < 0.001, ns—not significant ( p > 0.2) by Student’s T-test compared to wild type.

Article Snippet: Staining was performed using the following primary antibodies: mouse anti-Synapsin (3C11) 1:50 [ ] (Developmental Studies Hybridoma Bank, University of Iowa–DSHB – deposited by Buchner, E.); rabbit anti-Dlg 1:30,000 [ ]; mouse anti-Brp (nc82) 1:250 [ ] (deposited to DSHB by Buchner, E.); mouse anti-FasII (1D4) 1:900 [ ] (deposited to DSHB by Goodman, C.); mouse anti-GluRIIA (8B4D2) 1:500 [ ] (deposited to DSHB by Goodman, C.).

Techniques: Immunostaining, Expressing, Fluorescence, Western Blot, Quantitative Proteomics, Molecular Weight

Journal: PLoS Genetics

Article Title: C-terminal Src Kinase Gates Homeostatic Synaptic Plasticity and Regulates Fasciclin II Expression at the Drosophila Neuromuscular Junction

doi: 10.1371/journal.pgen.1005886

Figure Lengend Snippet: A) Values for mEPSP amplitude (gray) and quantal content (QC; white) normalized to genetic controls (dashed line) that lack a homeostatic challenge. FasII ; GluRIIA ; Csk triple mutants have intact homeostatic plasticity. (B-C) Representative electrophysiological traces for data shown in A. Scale bar for EPSP (mEPSP) traces: y = 5 mV (0.5 mV), x = 50 ms (1 s). (D-E’) Representative images of NMJs stained for FasII (green, gray channel) and Dlg (red) for (D, D’) Csk mutant and (E, E’) FasII , Csk double mutant NMJs. Scale bar = 10 μm. (F) Quantification of FasII staining intensities per synapse area for the genotypes shown in D-E’. (G) Western blot of FasII (DSHB 1D4 antibody) in protein extracts from whole third instar larvae. (H, I) Relative quantification of (H) total FasII intensity and (I) the intensity of the lowest molecular weight FasII band (the ‘third band’). Values are fold changes relative to wild type. * p < 0.05, ** p < 0.01, *** p < 0.001, ns—not significant ( p > 0.4) by Student’s T-test and for the bars in A, which are p -values from ANOVA (Tukey’s post-hoc) comparison between the three genotypes shown.

Article Snippet: Staining was performed using the following primary antibodies: mouse anti-Synapsin (3C11) 1:50 [ ] (Developmental Studies Hybridoma Bank, University of Iowa–DSHB – deposited by Buchner, E.); rabbit anti-Dlg 1:30,000 [ ]; mouse anti-Brp (nc82) 1:250 [ ] (deposited to DSHB by Buchner, E.); mouse anti-FasII (1D4) 1:900 [ ] (deposited to DSHB by Goodman, C.); mouse anti-GluRIIA (8B4D2) 1:500 [ ] (deposited to DSHB by Goodman, C.).

Techniques: Staining, Mutagenesis, Western Blot, Quantitative Proteomics, Molecular Weight, Comparison

Journal: Disease Models & Mechanisms

Article Title: Individual components of the SWI/SNF chromatin remodelling complex have distinct roles in memory neurons of the Drosophila mushroom body

doi: 10.1242/dmm.037325

Figure Lengend Snippet: Some SWI/SNF components are required for MBγ neuron remodelling. (A) Schematic diagram of MBγ neuron remodelling. APF, after pupae formation. The dashed line indicates the part of the MBγ lobe that is pruned. (B-D) Confocal projections showing MB neurons labelled with R14H06-Gal4 and UAS-mCD8::GFP . Controls expressing an RNAi against mCherry were compared to SWI/SNF knockdown RNAi lines for Bap60 ( UAS-Bap60 32503 ), Snr1 ( UAS-Snr1 32372 ) and E(y)3 [ UAS-e(y)3 32346 ]. Images were obtained for adults (B), third-instar larvae (C) and early pupae (D). FasII was labelled by immunohistochemistry. Scale bars: 50 µm. Arrows indicate the location of unpruned MBγ axons. For each genotype and developmental stage, we imaged a minimum of ten brains. Larval (C) and pupal (D) phenotypes were 100% penetrant. The penetrance of adult phenotypes is quantified in Fig. 3 .

Article Snippet: For immunohistochemistry, fixed brains were incubated overnight with the primary antibodies anti-FasII (1:25; DSHB, 1D1), anti-Brp (1:50; DSHB, nc82) and anti-EcR-B1 (1:25; DSHB, AD4.4), and the secondary antibody goat anti-mouse DyLight 594 (1:300).

Techniques: Expressing, Knockdown, Immunohistochemistry