Journal: Neurochemistry international

Article Title: Protein kinase C modulates synaptic vesicle acidification in a ribbon type nerve terminal in the retina

doi: 10.1016/j.neuint.2008.07.004

Figure Lengend Snippet: LysoTracker stains preferentially retinal bipolar terminals. (A) DIC image of a representative bipolar Mb cell (left). The same cell was stained with LT (100 nM) for 10 min (right). (B) Graphic showing increased LT fluorescence levels in cell terminals compared to cell bodies (*P < 0.05) after 10 min of LT exposure. n = 7 cells. Scale bar = 10 μM.

Article Snippet: Fluorescence imaging was achieved with a Zeiss water immersion objective (40×, 1.2 NA) or Zeiss oil immersion objective (63×, NA) coupled to a Zeiss LSM510 laser scanning confocal microscope (CEMEL-UFMG) or a Biorad MRC1024 confocal system (Department of Pharmacology, ICB-UFMG).

Techniques: Staining, Fluorescence

Journal: Neurochemistry international

Article Title: Protein kinase C modulates synaptic vesicle acidification in a ribbon type nerve terminal in the retina

doi: 10.1016/j.neuint.2008.07.004

Figure Lengend Snippet: PKA and CAMKII do not change LysoTracker fluorescence in retinal bipolar terminals. (A) Panel showing representative cell terminals stained with LT 100 nM (first panel) in association with NH4Cl 30 mM (second panel); SP-cAMP 100 nM (third panel), RP-cAMP 100 nM (fourth panel), and KN93 1 μM (fifth panel). Scale bar = 10 μM. Upper row, DIC images; lower row, fluorescence images. (B) Graphic showing that cells exposed to NH4Cl were less stained with LT compared to control (*P < 0.05; n = 8) whereas SP-cAMP (P > 0.05; n = 10), RP-cAMP (P > 0.05; n = 9) and KN93 (P > 0.05; n = 9) did not cause significant changes in fluorescence in retinal bipolar terminals if compared to their controls.

Article Snippet: Fluorescence imaging was achieved with a Zeiss water immersion objective (40×, 1.2 NA) or Zeiss oil immersion objective (63×, NA) coupled to a Zeiss LSM510 laser scanning confocal microscope (CEMEL-UFMG) or a Biorad MRC1024 confocal system (Department of Pharmacology, ICB-UFMG).

Techniques: Fluorescence, Staining, Control

Journal: Neurochemistry international

Article Title: Protein kinase C modulates synaptic vesicle acidification in a ribbon type nerve terminal in the retina

doi: 10.1016/j.neuint.2008.07.004

Figure Lengend Snippet: Activation of PKC increases LysoTracker fluorescence in retinal bipolar terminals. (A) Panel showing representative cell terminals of different cells stained with LT 100 nM (first panel) and exposed to PMA 100 nM (second panel); calphostin C 100 nM (third panel) and calphostin C + PMA (fourth panel). Scale bar = 10 μM. Upper row, DIC images; lower row, fluorescence images. (B) Bar graph showing that PMA increased LT fluorescence compared to control (*P < 0.05; n = 8) while cells exposed to calphostin C (P > 0.05; n = 9) and calphostin C + PMA (P > 0.05; n = 9) did not have their fluorescence changed if compared to their controls.

Article Snippet: Fluorescence imaging was achieved with a Zeiss water immersion objective (40×, 1.2 NA) or Zeiss oil immersion objective (63×, NA) coupled to a Zeiss LSM510 laser scanning confocal microscope (CEMEL-UFMG) or a Biorad MRC1024 confocal system (Department of Pharmacology, ICB-UFMG).

Techniques: Activation Assay, Fluorescence, Staining, Control

Journal: Neurochemistry international

Article Title: Protein kinase C modulates synaptic vesicle acidification in a ribbon type nerve terminal in the retina

doi: 10.1016/j.neuint.2008.07.004

Figure Lengend Snippet: LysoSensor staining of retinal bipolar terminals. (A) DIC image of a representative bipolar Mb cell (left). Same cell after staining with LS (100 nM) for 20 min (right). (B) Graphic showing increased LS fluorescence levels in cell terminals compared to cell bodies (*P < 0.05) after 20 min of dye exposure. n = 11 cells. Scale bar = 10 μM.

Article Snippet: Fluorescence imaging was achieved with a Zeiss water immersion objective (40×, 1.2 NA) or Zeiss oil immersion objective (63×, NA) coupled to a Zeiss LSM510 laser scanning confocal microscope (CEMEL-UFMG) or a Biorad MRC1024 confocal system (Department of Pharmacology, ICB-UFMG).

Techniques: Staining, Fluorescence

Journal: Neurochemistry international

Article Title: Protein kinase C modulates synaptic vesicle acidification in a ribbon type nerve terminal in the retina

doi: 10.1016/j.neuint.2008.07.004

Figure Lengend Snippet: LysoSensor staining is dependent on transmembrane electrochemical gradient (ΔμH+). (A) Representative cell terminals stained with LS 100 nM (first panel), pre-incubated with methylamine 10 nM (second panel), NH4Cl 30 mM (third panel) and bafilomycin 500 nM (fourth panel). Scale bar = 10 μM. Upper row, DIC images; lower row, fluorescence images. (B) Graphic showing that all cell terminals pre-incubated with these pharmacological agents for 10 min and maintained in the medium during LS staining showed a decrease in fluorescence compared to their controls (*P < 0.05; n = 8 for methylamine, n = 10 for NH4Cl, n = 10 for bafilomycin).

Article Snippet: Fluorescence imaging was achieved with a Zeiss water immersion objective (40×, 1.2 NA) or Zeiss oil immersion objective (63×, NA) coupled to a Zeiss LSM510 laser scanning confocal microscope (CEMEL-UFMG) or a Biorad MRC1024 confocal system (Department of Pharmacology, ICB-UFMG).

Techniques: Staining, Incubation, Fluorescence

Journal: Neurochemistry international

Article Title: Protein kinase C modulates synaptic vesicle acidification in a ribbon type nerve terminal in the retina

doi: 10.1016/j.neuint.2008.07.004

Figure Lengend Snippet: Activation of PKC increases synaptic vesicles acidification in retinal bipolar terminals. (A) Representative bipolar cells terminals stained with LS 100 nM (first panel), pre-incubated with PMA 100 nM (second panel); calphostin C 100 nM (third panel) and calphostin C + PMA (fourth panel). Upper row, DIC images; lower row, fluorescence images. Scale bar = 10 μM. (B) Bar graphic showing that cells exposed to PMA increased LS fluorescence (*P < 0.05; n = 9) while the group of cells exposed to calphostin C (P > 0.05; n = 9), calphostin C + PMA (P > 0.05; n = 9), staurosporine (P > 0.05; n = 10), staurosporine + PMA (P > 0.05; n = 9) and 4alpha-PMA (P > 0.05; n = 8) did not cause a significant change in fluorescence compared to their controls.

Article Snippet: Fluorescence imaging was achieved with a Zeiss water immersion objective (40×, 1.2 NA) or Zeiss oil immersion objective (63×, NA) coupled to a Zeiss LSM510 laser scanning confocal microscope (CEMEL-UFMG) or a Biorad MRC1024 confocal system (Department of Pharmacology, ICB-UFMG).

Techniques: Activation Assay, Staining, Incubation, Fluorescence