bioimagesUnknownSandra DieniThe Journal of Cell Biologyhttps://www.ebi.ac.uk/biostudies/studies/S-JCBD-201201038JCB

Electron micrograph of an ultrathin section of a wild-type mouse stratum lucidum pre-labeled with anti-BDNF immunogold particles.

Pro-BDNF antibodies (green) do not produce an immunoreactive signal in the CA3 region of a cbdnf-/- mouse hippocampus.

Anti-pro-BDNF-Mab5H8-IR (green) labeling is markedly increased in the hilus of the dentate gyrus in a Bsn mouse mutant.

Electron micrograph of a cbdnf-/- mouse dendritic profile in the stratum lucidum labeled with anti-pro-BDNF-immunogold antibodies.

Pro-BDNF antibodies (green) do not produce an immunoreactive signal in the dentate gyrus region of a cbdnf-/- mouse hippocampus.

An electron micrograph of anti-BDNF-immunogold labeling of sections from cbdnf-/- mice yielded non-specific background labeling.

Anti-BDNF-IR (red) labeling is markedly increased in the CA3 area of a Bsn mouse mutant.

Low power confocal z-stack of the dentate gyrus of a wild-type mouse hippocampus. The same subset of granule cells co-expresses BDNF-IR (red) and pro-BDNF-IR (green).

Electron micrograph showing the Golgi complex of a wild-type mouse CA3 neuron labeled with anti-pro-BDNF immunogold. Clusters of gold grains aggregate around the cisterns of the Golgi apparatus.

Anti-BDNF-IR (red) labeling is markedly increased in the hilus of the dentate gyrus of a Bsn mouse mutant.

High resolution confocal z-slice of the stratum radiatum in the CA1 region of a BDNF-Myc mouse. Overlay of anti-Myc (red) and anti-synaptopodin (green) labeling shows presynaptic localization of BDNF, with close apposition of Myc-IR with synpo-positive spines.

Confocal z-slice of a wild-type mouse stratum lucidum labeled with antibodies to Myc (red) and VGLUT-1 (green).

Myc-IR (red) and CCK-IR (green) in presynaptic terminals within a wild-type mouse stratum lucidum. The merged image indicates complete segregation of the two peptides in the mossy fiber projection.

A small proportion of mossy fiber boutons in the wild-type mouse stratum lucidum co-expresses Myc-IR (red) and Met-enk-IR (green). However, in the merged image (shown), Myc- and Met-enk-IR puncta do not overlap.

Electron micrograph of mossy fiber boutons from a Bsn mouse mutant section pre-labeled with BDNF immunogold.

In sections processed for anti-BDNF immunogold, gold-labeled vesicles were observed in electron micrographs of wild-type mouse unmyelinated mossy fiber axons.

Electron micrograph of a mossy fiber bouton from a wild-type mouse section pre-labeled with anti-pro-BDNF immunogold.

Electron micrograph of a wild-type mouse dendrite labeled with pro-BDNF-immunogold antibodies.

Electron micrograph of a wild-type mouse dendritic profile in the stratum lucidum labeled with anti-pro-BDNF-immunogold antibodies.

Immunoprecipitation with anti-BDNF antisera and Western blot analysis with anti-BDNF N-20 antibody demonstrating the recovery of wild-type recombinant pro-BDNF (upper bands, lanes 2,5) and mature BDNF (lower bands, lanes 2,5) from mouse hippocampal lysates in the absence (lane 2) or presence (lane 5) of protease inhibitors. Lanes 1, 3 and 4 represent control samples (1 = +wild-type HIP only; 3 = +protease inhibitors, +recombinant BDNF; 4 = +wild-type HIP, +protease inhibitors).

Low power images of a wild-type mouse dentate gyrus labeled with anti-BDNF (red) and anti-pro-BDNF-(green) antibodies.

High magnification view of the dentate gyrus region of a cbdnf-/- mouse hippocampus stained with anti-BDNF antibodies (red). Note the absence of immunoreactive signals in all cellular and neuropil layers.

No BDNF-positive secretory vesicles were found by immunogold labeling in a section from a cbdnf-/- mice visualized in this electron micrograph.

In the stratum lucidum of a Bsn mouse mutant, an increased density of labeled profiles was observed, with BDNF-IR (green) labeling showing virtually no overlap with labeling for the post-synaptic marker MAP-2 (red).

Low power view of a wild-type mouse hippocampal section stained with anti-BDNF antibodies (red).

A high resolution confocal z-slice shows co-localization of BDNF-IR (red) and pro-BDNF-IR (green) labeling at the border of stratum radiatum and stratum lacunosum-moleculare in a section from a Bsn mutant mouse.

No pro-BDNF-positive secretory vesicles were found by immunogold labeling in this electron micrograph of a section from a cbdnf-/- mice.

Anti-BDNF-positive granule cells in a Bsn mouse mutant shows labeling confined to the cell soma and no detectable dendritic staining in the inner molecular layer.

Electron micrograph of a Bsn mouse mutant dendrite labeled with anti-BDNF immunogold.

Low power image of a wild-type mouse CA3 region labeled with anti-pro-BDNF-Mab5H8 (green) antibodies.

No specific staining is observed in the dentate gyrus region of a wild-type mouse hippocampus section treated with anti-Myc antibodies (red).

Electron micrograph of a cbdnf-/- mouse dendritic profile in the stratum lucidum labeled with anti-BDNF-immunogold antibodies.

Merged confocal z-slice of a wild-type mouse stratum lucidum labeled with antibodies to Myc (red) and Synaptophysin (green).

Electron micrograph of a wild-type mouse dendrite labeled with anti-BDNF immunogold.

Anti-pro-BDNF-Mab5H8-positive granule cells (green) in a Bsn mouse mutant shows labeling confined to the cell soma and no detectable dendritic staining in the inner molecular layer.

Electron micrograph of a Bsn mouse mutant dendrite labeled with pro-BDNF immunogold.

Shown is Myc-IR (red) and MAP-2-IR (green) labeling in a confocal z-stack from a wild-type mouse CA3 region highlighting Myc-positive neuronal somata in the PCL and mossy fiber bouton profiles in the stratum lucidum. No overlap between Myc and MAP2 labeling can be seen.

Electron micrograph of a mossy fiber bouton from a Bsn mouse mutant section pre-labeled with anti- pro-BDNF immunogold.

Low power image of a wild-type mouse CA3 region labeled with anti-BDNF antibodies (red).

Anti-pro-BDNF-Mab5H8-IR (green) labeling is markedly increased in the CA3 area of a Bsn mouse mutant.

A confocal stack of a BDNF-(red) and pro-BDNF (green)-immunoreactive granule cell counterstained with DAPI (blue) from a wild-type mouse hippocampus. Labeling concentrated at the cell apex in distinct clusters, with partial overlap of the BDNF and pro-BDNF signals.

Electron micrograph of a mossy fiber bouton from a wild-type mouse section pre-labeled with anti-BDNF immunogold.

BDNF-IR (red) and pro-BDNF-IR (green) labeling in a Bsn mouse mutant shows a laminar-specific increase in staining at the border of stratum radiatum and stratum laconosum-moleculare.

Electron micrograph of a wild-type mouse dendritic profile in the stratum lucidum labeled with anti-BDNF-immunogold antibodies.

In stratum lucidum of a Bsn mouse mutant, an increased density of labeled profiles was observed, with BDNF-IR (red) labeling showing virtually no overlap with labeling for the post-synaptic marker Synaptopodin (green).

High resolution confocal z-stack of a wild-type mouse dentate gyrus labeled with antibodies against BDNF (red), pro-BDNF (green) and Arc/Arg3.1 (blue).

No specific staining in the CA3 region of a wild-type mouse hippocampus section treated with anti-Myc antibodies (red).

Both pro-BDNF and its pro-peptide were detected by Western blotting of mouse hippocampal lysates immunoprecipitated with anti-pro-BDNF antibody and detected with anti-pro-BDNF monoclonal antibody 5H8 in wildtype (lane 1) but not in cbdnf-/- mouse samples (lane 2). Cleavage-resistant pro-BDNF (lane 3), recombinant BDNF pro-peptide (lanes 4 & 5) and recombinant non-glycosylated pro-peptide (lane 6) were used as molecular weight markers.

A Bdnf-Myc-expressing mouse hippocampus section treated with anti-Myc antibodies (red) shows a similar staining pattern to that produced by anti-BDNF antibodies.

Polyclonal pro-BDNF antibodies (green) yield a similar pattern to that of anti-BDNF in a wild-type mouse hippocampus section.

Co-labeling of the Bsn mouse mutant CA1 region with antibodies against BDNF (red) and Synaptopodin (green) reveals a segregation of signals at the border of stratum radiatum and stratum lacunosum-moleculare.

Co-labeling of a Bsn mouse mutant CA1 region with antibodies against BDNF (red) and GFAP (green) shows a lack of BDNF-IR in astrocytes in a confocal z-stack.

High resolution confocal z-stack of a single CA3 neuron from a wild-type mouse hippocampus labeled with antibodies against Myc (red), pro-BDNF (green) and GM130 (light blue).

Electron micrograph of an anti-pro-BDNF immunogold-labeled wild-type mouse mossy fiber bouton containing cluster-labeled secretory vesicles.

Electron micrograph showing anti-BDNF-immunogold labeling of a mouse presynaptic terminal in the stratum radiatum of CA1.

Immunoprecipitation with anti-BDNF antisera and Western blot analysis with anti-BDNF N20 antibody of hippocampal lysates from Bsn mouse mutants revealed a relatively similar increase (~3 fold) in the levels of BDNF and pro-BDNF (lane 3) compared to wild-type lysates (lanes 1 and 2). Recombinant mouse BDNF (lanes 4-6, 8, 10) was used as a molecular weight marker.

Confocal z-slice of area CA3 of a wild-type mouse hippocampus, demonstrating BDNF-IR (red) and pro-BDNF-IR (green) in a subset of CA3 pyramidal cell somata. DAPI staining is shown in blue.

Confocal z-stack of a wild-type mouse granule cell co-expressing Myc-IR (red) and Met-enk-IR (green). Puncta are similarly distributed throughout the cell but show little overlap.

Immunoprecipitation with anti-pro-BDNF antisera and Western blot analysis with anti-pro-BDNF monoclonal antibody 5H8 of hippocampal lysates from Bsn mouse mutants revealed a relatively similar increase (~3 fold) in the levels of pro-peptide and pro-BDNF (lane 2), compared to wild-type tissues (lane 1). Recombinant BDNF pro-peptide (lanes 3,4,5) and cleavage-resistant pro-BDNF (lanes 3,4,6) were used as molecular weight markers.

High resolution confocal z-slice of a wild-type mouse stratum lucidum showing Myc-IR (red) and pro-BDNF-IR (green) in the same subset of mossy fiber boutons.

Higher magnification view of the CA3 region of a cbdnf-/- mouse hippocampus stained with anti-BDNF antibodies (red). Note the absence of immunoreactive signals in all cellular and neuropil layers.

Electron micrograph showing immunogold labeling of an anti-pro-BDNF-positive vesicle within a wild-type mouse mossy fiber that can been seen terminating into a bouton.

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