pan α n type ca 2 α 1b polyclonal antibody  (Santa Cruz Biotechnology)

Journal: PLoS ONE

Article Title: TRP, TRPL and Cacophony Channels Mediate Ca 2+ Influx and Exocytosis in Photoreceptors Axons in Drosophila

doi: 10.1371/journal.pone.0044182

Figure Lengend Snippet: ( A–C ) Confocal images of the visual system from GMR-Gal4/UAS-G-CaMP transgenic flies showing the expression pattern of the Ca 2+ indicator protein, G-CaMP, in Drosophila photoreceptors. ( D ) Pseudocolor fluorescence images illustrating Ca 2+ increments upon depolarization induced by high-K + (90 mM); effect of the cacophony blocker PLTX-II on G-CaMP/Ca 2+ fluorescence changes evoked by high-K + . ( E ) Quantification of the G-CaMP/Ca 2+ fluorescence changes illustrated in (D).

Article Snippet: The α-TRPL polyclonal antibody AB5912 was obtained from Chemicon and the pan α-N-type Ca 2+ α-1B polyclonal antibody (L-17), from Santa Cruz .

Techniques: Transgenic Assay, Expressing, Fluorescence

Journal: PLoS ONE

Article Title: TRP, TRPL and Cacophony Channels Mediate Ca 2+ Influx and Exocytosis in Photoreceptors Axons in Drosophila

doi: 10.1371/journal.pone.0044182

Figure Lengend Snippet: ( A ) Z-projections of 10 confocal optical sections (Δz = 0.3 mm) showing the fluorescence of FM4-64-loaded boutons in the lamina from: ( a ) wt not exposed to high-K + (control); ( b ) wt after depolarization by high-K + ; ( c ) wt depolarized by high-K + in the presence of PLTX-II (100 nM); ( d ) wt depolarized by high-K + in 0-Ca 2+ external solution; ( e ) trp depolarized by high-K + ; ( f ) trpl depolarized by high-K + ; ( g ) trpl;trp depolarized by high-K + . ( B ) Bouton labeling induced by the Ca 2+ ionophore, A-23187 (250 nM) in wt (left) and trpl;trp (right) in regular Ringer (5 mM K + ). ( C ) Quantification of the number of labeled synaptic boutons for the different conditions shown in A. ( D ) Bouton labeling induced by high-K + in slices from cac TS mutants pre-incubated in chilled Ringer (left, Control) or at 37°C (center) for 10 minutes. Right: bouton labeling at 37°C in slices from wt flies. (E) Quantification of the number of synaptic boutons for the conditions shown in D. Bars: mean ± SEM, calculated from z-projections of 10 images. Size: x/y/z = 36/36/0.3 µm 3 . * p<0.05: respect to wt high-K + labeling.

Article Snippet: The α-TRPL polyclonal antibody AB5912 was obtained from Chemicon and the pan α-N-type Ca 2+ α-1B polyclonal antibody (L-17), from Santa Cruz .

Techniques: Fluorescence, Control, Labeling, Incubation

Journal: PLoS ONE

Article Title: TRP, TRPL and Cacophony Channels Mediate Ca 2+ Influx and Exocytosis in Photoreceptors Axons in Drosophila

doi: 10.1371/journal.pone.0044182

Figure Lengend Snippet: Slices were bathed with 0-Ca 2+ solution supplemented with 10 µM Thg (Thg(0Ca 2+ )) during 8 min to produce internal Ca 2+ stores (ER) depletion. Afterwards the slices were returned to regular Ringer. ( A, B ) Z-projections of 4 confocal images were obtained from the lamina in slices preloaded with Rhod-2 and treated with the depletion protocol. Rhod-2 fluorescence in wt (A) or trpl;trp (B) slices in regular Ringer (left), during application of Thg(0Ca 2+ ) solution (center) and after regular Ringer was restored (right). ( C–D ) Z-projections of 10 confocal optical sections (Δz = 0.3 mm) showing representative bouton loading of FM4-64 in wt (C) and trpl;trp (D) slices, upon returning to regular Ringer after ER depletion. ( E ) Quantification of Rhod-2 fluorescence change after restoring Ringer in wt (A) and trpl;trp (B). ( F ) Quantification of FM4-64 bouton labeling induced by the depletion protocol (C and D, denoted by “0Ca 2+ +Thg”) or upon returning to Ringer after treatment with 0-Ca 2+ solution without Thg (“0Ca 2+ ”). Bars: mean ± SEM calculated in z-projections of 10 images. Size: x/y/z = 36/36/0.3 µm 3 . * p<0.05 with respect to wt. Pseudocolor scale in arbitrary units.

Article Snippet: The α-TRPL polyclonal antibody AB5912 was obtained from Chemicon and the pan α-N-type Ca 2+ α-1B polyclonal antibody (L-17), from Santa Cruz .

Techniques: Fluorescence, Labeling

Journal: PLoS ONE

Article Title: TRP, TRPL and Cacophony Channels Mediate Ca 2+ Influx and Exocytosis in Photoreceptors Axons in Drosophila

doi: 10.1371/journal.pone.0044182

Figure Lengend Snippet: A graded depolarization from the soma reaches the axon terminals (1) opening cacophony, which allows the Ca 2+ influx (2) that triggers exocytosis. Additional mechanisms complement or amplify the Ca 2+ signal: Ca 2+ release from the endoplasmic reticulum (ER) (3), PLC activation (4) and Ca 2+ influx through TRP/TRPL channels (5). Internal Ca 2+ release could be due to ryanodine receptor (RyR) activation, a mechanism termed Ca 2+ -induced Ca 2+ release, or mediated by IP 3 receptor (IP 3 R) opening as a result of PLC activity. TRP/TRPL working as store-operated channels (SOCs) contribute Ca 2+ to exocytosis and could also be modulated by PLC-dependent lipid changes. The massive raise of Ca 2+ from these multiple pathways allows extremely fast exocytosis at the synaptic terminal. In green are displayed the components and pathways shown by us to be involved in vesicle exocytosis. The broken lines denote hypothetical pathways.

Article Snippet: The α-TRPL polyclonal antibody AB5912 was obtained from Chemicon and the pan α-N-type Ca 2+ α-1B polyclonal antibody (L-17), from Santa Cruz .

Techniques: Activation Assay, Activity Assay