Journal: PLoS ONE

Article Title: Cyclical and Patch-Like GDNF Distribution along the Basal Surface of Sertoli Cells in Mouse and Hamster Testes

doi: 10.1371/journal.pone.0028367

Figure Lengend Snippet: Rotating 3D reconstruction showing the close co-localization of a GFRα1-positive spermatogonial cell with immunoreactive GDNF-positive deposits in the basal compartment of seminiferous tubule in hamster testes. PFA-fixed seminiferous tubule fragments were double-stained with anti-GDNF (green) and GFRα1 (red) antibodies (DAPI, blue) without any permeabilization steps, and then analyzed to reconstruct a three-dimensional image using an Olympus FluoView confocal laser microscope (FV10i; Olympus, Japan) in combination with Volocity software (Mitani Sangyo, Japan) (see also Fig. 7 ).

Article Snippet: Without any permeabilization steps using methanol and detergent (Tween20/Triton X-100), the seminiferous tubule fragments were incubated with rabbit anti-GDNF (1∶200 dilution) and goat anti-GFRα1 (1∶100 dilution; R&D Systems)/goat anti-mouse c-kit (1∶100 dilution in mice, 1∶20 in hamsters; R&D Systems) antibodies at 4°C for 12 h. After being washed with PBS, the samples were incubated with Alexa-488/594 conjugated secondary antibodies, including DAPI, at room temperature for 2 h. After counter-staining with DAPI, the samples were analyzed under Olympus fluorescent microscope (BX51N-34-FL2) and stereomicroscope (SZX16 plus U-LH100HG) systems and Olympus FluoView confocal laser microscope (FV10i; Olympus, Japan) in combination with Volocity software (Mitani Sangyo, Japan).

Techniques:

Journal: PLoS ONE

Article Title: Cyclical and Patch-Like GDNF Distribution along the Basal Surface of Sertoli Cells in Mouse and Hamster Testes

doi: 10.1371/journal.pone.0028367

Figure Lengend Snippet: Rotating 3D reconstruction showing the close co-localization of a GFRα1-positive spermatogonial cell with immunoreactive GDNF-positive deposits in the basal compartment of seminiferous tubule in mouse testes. PFA-fixed seminiferous tubule fragments were double-stained with anti-GDNF (green) and GFRα1 (red) antibodies (DAPI, blue) without any permeabilization steps, and then analyzed to reconstruct a three-dimensional image using an Olympus FluoView confocal laser microscope (FV10i; Olympus, Japan) in combination with Volocity software (Mitani Sangyo, Japan) (see also Fig. 7 ).

Article Snippet: Without any permeabilization steps using methanol and detergent (Tween20/Triton X-100), the seminiferous tubule fragments were incubated with rabbit anti-GDNF (1∶200 dilution) and goat anti-GFRα1 (1∶100 dilution; R&D Systems)/goat anti-mouse c-kit (1∶100 dilution in mice, 1∶20 in hamsters; R&D Systems) antibodies at 4°C for 12 h. After being washed with PBS, the samples were incubated with Alexa-488/594 conjugated secondary antibodies, including DAPI, at room temperature for 2 h. After counter-staining with DAPI, the samples were analyzed under Olympus fluorescent microscope (BX51N-34-FL2) and stereomicroscope (SZX16 plus U-LH100HG) systems and Olympus FluoView confocal laser microscope (FV10i; Olympus, Japan) in combination with Volocity software (Mitani Sangyo, Japan).

Techniques: