Journal: Brain Sciences

Article Title: Neonatal Handling Positively Modulates Anxiety, Sensorimotor Gating, Working Memory, and Cortico-Hippocampal Neuroplastic Adaptations in Two Genetically Selected Rat Strains Differing in Emotional and Cognitive Traits

doi: 10.3390/brainsci15080776

Figure Lengend Snippet: Western blot analysis of the BDNF ( A , B ), trkB ( C , D ), and the PSA-NCAM ( E , F ), in the prefrontal (prelimbic/infralimbic) cortex of the RHA and the RLA rats, either untreated controls (CTRL) or treated with neonatal handling (NH). The values represent the densitometric analysis of the BDNF/GAPDH ( B ), trkB/GAPDH ( D ), and the PSA-NCAM/GAPDH ( F ) band grey optical density (O.D.) ratios. The bars denote the mean ± S.E.M. of 9–10 rats, in each experimental group. ***: p < 0.001 (post hoc Duncan’s multiple range test).

Article Snippet: Molecular weight (mw) standards (Precision Plus Protein Western C Standards, Cat# 161–0376, Bio-Rad, Hercules, CA, USA) and recombinant human BDNF protein (rhBDNF) (Cat# B-257, Alomone Labs, Jerusalem, Israel) were run in parallel.

Techniques: Western Blot

Journal: Brain Sciences

Article Title: Neonatal Handling Positively Modulates Anxiety, Sensorimotor Gating, Working Memory, and Cortico-Hippocampal Neuroplastic Adaptations in Two Genetically Selected Rat Strains Differing in Emotional and Cognitive Traits

doi: 10.3390/brainsci15080776

Figure Lengend Snippet: Western blot analysis of the BDNF ( A , B ), trkB ( C , D ), and the PSA-NCAM ( E , F ), in the anterior cingulate cortex of the RHA and the RLA rats, either untreated controls (CTRL) or treated with neonatal handling (NH). The values represent the densitometric analysis of the BDNF/GAPDH ( B ), trkB/GAPDH ( D ), and the PSA-NCAM/GAPDH ( F ) band grey optical density (O.D.) ratios. The bars denote the mean ± S.E.M. of 9–10 rats, in each experimental group. **: p < 0.01 (post hoc Duncan’s multiple range test).

Article Snippet: Molecular weight (mw) standards (Precision Plus Protein Western C Standards, Cat# 161–0376, Bio-Rad, Hercules, CA, USA) and recombinant human BDNF protein (rhBDNF) (Cat# B-257, Alomone Labs, Jerusalem, Israel) were run in parallel.

Techniques: Western Blot

Journal: Brain Sciences

Article Title: Neonatal Handling Positively Modulates Anxiety, Sensorimotor Gating, Working Memory, and Cortico-Hippocampal Neuroplastic Adaptations in Two Genetically Selected Rat Strains Differing in Emotional and Cognitive Traits

doi: 10.3390/brainsci15080776

Figure Lengend Snippet: Western blot analysis of the BDNF ( A , B ), trkB ( C , D ), and the PSA-NCAM ( E , F ), in the ventral hippocampus of the RHA and the RLA rats, either untreated controls (CTRL) or treated with neonatal handling (NH). The values represent the densitometric analysis of the BDNF/GAPDH ( B ), trkB/GAPDH ( D ), and the PSA-NCAM/GAPDH ( F ) band grey optical density (O.D.) ratios. The bars denote the mean ± S.E.M. of 9–10 rats, in each experimental group. *: p < 0.05; **: p < 0.02 (post hoc Duncan’s multiple range test).

Article Snippet: Molecular weight (mw) standards (Precision Plus Protein Western C Standards, Cat# 161–0376, Bio-Rad, Hercules, CA, USA) and recombinant human BDNF protein (rhBDNF) (Cat# B-257, Alomone Labs, Jerusalem, Israel) were run in parallel.

Techniques: Western Blot

Journal: Brain Sciences

Article Title: Neonatal Handling Positively Modulates Anxiety, Sensorimotor Gating, Working Memory, and Cortico-Hippocampal Neuroplastic Adaptations in Two Genetically Selected Rat Strains Differing in Emotional and Cognitive Traits

doi: 10.3390/brainsci15080776

Figure Lengend Snippet: Western blot analysis of the BDNF ( A , B ), trkB ( C , D ), and the PSA-NCAM ( E , F ), in the dorsal hippocampus of the RHA and the RLA rats, either untreated controls (CTRL) or treated with neonatal handling (NH). The values represent the densitometric analysis of the BDNF/GAPDH ( B ), trkB/GAPDH ( D ), and the PSA-NCAM/GAPDH ( F ) band grey optical density (O.D.) ratios. The bars denote the mean ± S.E.M. of 9–10 rats, in each experimental group.

Article Snippet: Molecular weight (mw) standards (Precision Plus Protein Western C Standards, Cat# 161–0376, Bio-Rad, Hercules, CA, USA) and recombinant human BDNF protein (rhBDNF) (Cat# B-257, Alomone Labs, Jerusalem, Israel) were run in parallel.

Techniques: Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Fingolimod Modulates Dendritic Architecture in a BDNF-Dependent Manner

doi: 10.3390/ijms21093079

Figure Lengend Snippet: Fingolimod-phosphate (FTY720-P) regulates neuronal architecture in a BDNF-dependent manner. ( A ) Representative Neurolucida tracings, used to perform the Sholl analysis of dendritic complexity for feGFP expressing neurons treated for 24h with: DMSO (black), 2nM FTY720-P (gray), DMSO + TrkBFc (dark blue) and 2nM FTY720-P + TrkBFc (light blue). Scale bar: 100 μm. ( B ) Sholl analysis displayed as number of dendritic intersections against distance from the cell body for DMSO (black), 2nM FTY720-P (gray), DMSO + TrkBFc (dark blue) and 2nM FTY720-P + TrkBFc (light blue) and ( C ) total dendritic complexity of all treatment groups. F value in ( B ) refers to comparison between all the 4 treatment groups. ( D ) Dendritic spine densities for DMSO (black), 2nM FTY720-P (gray), DMSO + TrkBFc (dark blue) and 2nM FTY720-P + TrkBFc (light blue) calculated using segments of secondary dendritic branches as shown in ( E ) feGFP panel. The Syn panel displays the corresponding staining of the pre-synaptic marker SynapsinI/II and the merge panel shows the images with overlapping SynapsinI/II puncta (red, pre-synapse) to its matching feGFP dendrite segment (green, post-synapse). The arrows point to coinciding puncta, indicative of mature synapse between the post and pre-synaptic compartments. Scale bar: 5μm. ( F ) The graph compares the fraction of SynapsinI/II positive feGFP labelled spines for DMSO (black), 2nM FTY720-P (gray), DMSO + TrkBFc (dark blue) and 2nM FTY720-P + TrkBFc (light blue). ( G ) Representative fields of view (FOV) of the DMSO, 2nM FTY720-P, DMSO + TrkBFc and 2nM FTY720-P + TrkBFc treated hippocampal cultures stained for c-fo s . Scale bar: 100μm. ( H ) Quantification of the proportion of c-Fos expressing neurons represented as normalized fraction for DMSO (black), 2nM FTY720-P (gray), DMSO + TrkBFc (dark blue) and 2nM FTY720-P + TrkBFc (light blue) treated cultures. All graphs represent data as mean + SEM. Numbers in the bars show either total number of neurons or of FOV analyzed, obtained from ≥3 sets of independent experiments. Two-way ANOVA followed by Bonferroni post-hoc test was used in ( B ). For ( C , D , F ) and ( H ) one-way ANOVA with Bonferroni post-hoc was used. Denotations for significance are * p < 0.05, ** p < 0.01, **** p < 0.0001.

Article Snippet: Recombinant Human BDNF protein (R&D systems, Minneapolis, MN, USA) was dissolved in sterile PBS with 0.1% BSA at a concentration of 50 ng/μL.

Techniques: Expressing, Comparison, Staining, Marker

Journal: International Journal of Molecular Sciences

Article Title: Fingolimod Modulates Dendritic Architecture in a BDNF-Dependent Manner

doi: 10.3390/ijms21093079

Figure Lengend Snippet: Treatment with the non-phosphorylated Fingolimod (FTY720) modulates neuronal architecture. ( A ) | Representative Neurolucida tracings from feGFP positive hippocampal neurons used for the Sholl analysis from cultures treated either with DMSO or 10nM FTY20 for 24 h. Scale bar: 100μm. ( B ) | Dendritic complexity shown by the number of dendritic intersections plotted against the distance from the soma for DMSO (black) and FTY720 (blue) treated neurons. The F value shows the statistical comparison between the two groups. The inset graph represents total dendritic complexity upon treatment with DMSO (black) and FTY720 (blue). ( C ) | Total dendritic length for DMSO (black) and FTY720 (blue) treated neurons. ( D ) | Representative stretches from dendrites of eGFP transfected hippocampal neurons showing dendritic spine protrusions, treated either with DMSO or FTY720 for 24h. Scale bar: 5μm. ( E ) | The graph shows dendritic spine density for DMSO (black) and FTY720 (blue) treated neurons . ( F ) | Representative images of fields of view (FOV) from primary hippocampal cultures stained with anti phospho-ERK1/2 antibody, 30 min post-application of one of the following: DMSO, 10nM FTY720, DMSO_100, 100nM FTY720, DMSO_100 + TrkB-Fc, 100nM FTY720 + TrkB-Fc or 40ng recombinant BDNF protein as a positive control. Scale bar: 100μm. ( G ) | The graph displays the fraction of pERK1/2 expressing neurons relative to the total number of MAP2 + neurons. The data is normalized to the respective controls and compared between the different treatment groups: DMSO (black), 10nM FTY720 (light blue solid), DMSO100 (gray), 100nM FTY720 (dark blue solid), 40ng recombinant BDNF (magenta), DMSO100 + TrkB-Fc (gray open), 100nM FTY720 + TrkB-Fc (dark blue open). All data is plotted as mean + SEM. Numbers in the bars show total number of neurons or FOV analyzed, obtained from ≥3 sets of independent experiments. Two-way ANOVA followed by Bonferroni post-hoc test was used in ( B ). For ( B ) total intersections, ( C ) and ( E ) unpaired Student’s t-test and for ( G ) one-way ANOVA with Bonferroni post-hoc was used. Denotations for significance are * p < 0.05, *** p < 0.001, **** p < 0.0001.

Article Snippet: Recombinant Human BDNF protein (R&D systems, Minneapolis, MN, USA) was dissolved in sterile PBS with 0.1% BSA at a concentration of 50 ng/μL.

Techniques: Comparison, Transfection, Staining, Recombinant, Positive Control, Expressing

Journal: The Journal of Neuroscience

Article Title: Local and Target-Derived Brain-Derived Neurotrophic Factor Exert Opposing Effects on the Dendritic Arborization of Retinal Ganglion CellsIn Vivo

doi: 10.1523/jneurosci.22-17-07639.2002

Figure Lengend Snippet: Figure 1. Altering endogenous retinal and tectal BDNF levels in vivo. Diagrams representing a transverse view of a Xenopus tadpole brain and eye illustrate experimental procedures (see Materials and Methods). RGCs are depicted in red, relative endogenous BDNF expression levels (Cohen-Cory et al., 1996) are depicted in blue, and exogenously applied factors are depicted in green. A, Effects of altered tectal neurotrophins on RGC dendritic arborization. Control, anti-BDNF, or BDNF-treated green fluorescent microspheres were injected into the stage 38 tadpole tectum. At stage 42, RGCs were retrogradely labeled by injecting rhodamine–dextran in the contralateral tectum. At stage 45, dendritic morphologies of double-labeled RGCs were evaluated. A low-power view of a tadpole eye shows green fluorescent microspheres retrogradely transported to the retinal ganglion cell layer, where a rhodamine–dextran-labeled RGC soma can also be visualized (lines denote lens and eye periphery). Scale bar, 50 m. A single-plane, high-power view of a stage 45 retina reveals a rhodamine–dextran-labeled RGC with internalized green fluorescent microspheres. Scale bar, 5 m. B, Effects of altered retinal neurotrophins on RGC dendritic arborization. Control, anti-BDNF, or BDNF-treated microspheres were injected into the stage 38 tadpole retina, and then RGCs were retrogradely labeled at stage 42. The low-power view shows rhodamine–dextran-labeled RGCs and green fluorescent microspheres restricted within the tadpole eye. Scale bar, 200 m. The single-plane, high-power view of a stage 45 retina reveals the morphology of a rhodamine– dextran-labeled RGC surrounded by green fluorescent microspheres. Scale bar, 5 m. C, Effects of altered retinal neurotrophins on RGC axonal arborization in the tectum. Control, anti-BDNF, or BDNF-treated microspheres were injected into the stage 43 tadpole retina, and the morphology of DiI- or YFP-labeled RGC axon arbors was visualized 24 and 48 hr later. Confocal microscope images of a control RGC axon at 0 and 24 hr demonstrate normal RGC axon arborization dynamics. Scale bar, 20 m.

Article Snippet: Recombinant human BDNF (rhBDNF) was kindly provided by Amgen (Thousand Oaks, CA), recombinant human neurotrophin-4 (NT-4) was generously provided by Genentech (South San Francisco, CA), and anti-rhBDNF neutralizing antibody (mouse IgG1) was obtained from R & D Systems (Minneapolis, MN).

Techniques: In Vivo, Expressing, Control, Injection, Labeling, Microscopy

Journal: The Journal of Neuroscience

Article Title: Local and Target-Derived Brain-Derived Neurotrophic Factor Exert Opposing Effects on the Dendritic Arborization of Retinal Ganglion CellsIn Vivo

doi: 10.1523/jneurosci.22-17-07639.2002

Figure Lengend Snippet: Figure 2. Tectal BDNF retrogradely enhances RGC dendritic arborization. To determine whether tectal BDNF influences RGC dendritic arborization within the retina, tadpoles received tectal injections of microspheres treated with control, BDNF, or anti-BDNF function-blocking antibodies. Microsphere-containing neurons colabeled with rhodamine–dextran were analyzed morphologically (Fig. 1A). A, Image reconstructions of two rhodamine-labeled RGCs with simple and complex dendritic arbors illustrate differences in dendritic arbor morphologies. B, Images of RGC dendritic arbors reveal that increasing tectal BDNF enhances RGC dendritic arborization, whereas neutralizing endogenous tectal BDNF with function-blocking antibodies reduces RGC dendritic arborization. C, Quantitative analysis reveals that primary dendrite number, branch tip number, branch tips per primary dendrite, and overall dendritic length were significantly enhanced by increasing tectal BDNF and reduced by injecting anti-BDNF into the optic tectum. Scale bar, 5 m. Error bars indicate SEM. p 0.05; p 0.01; p 0.001.

Article Snippet: Recombinant human BDNF (rhBDNF) was kindly provided by Amgen (Thousand Oaks, CA), recombinant human neurotrophin-4 (NT-4) was generously provided by Genentech (South San Francisco, CA), and anti-rhBDNF neutralizing antibody (mouse IgG1) was obtained from R & D Systems (Minneapolis, MN).

Techniques: Control, Blocking Assay, Labeling

Journal: The Journal of Neuroscience

Article Title: Local and Target-Derived Brain-Derived Neurotrophic Factor Exert Opposing Effects on the Dendritic Arborization of Retinal Ganglion CellsIn Vivo

doi: 10.1523/jneurosci.22-17-07639.2002

Figure Lengend Snippet: Figure 4. Retinal BDNF inhibits RGC den- dritic arborization in a dose-dependent man- ner. To determine whether RGCs are sensi- tive to the concentration of BDNF in the retina, Xenopus retinas were microinjected with 1–100 ng/l BDNF or control micro- spheres at the onset of dendritic arborization. Quantitative measures of dendritic arbor morphology revealed a dose-dependent re- sponse to BDNF. The highest concentration of BDNF most dramatically decreased pri- mary dendrite number, branch tip number, tips per dendrite, and dendrite length versus control. Error bars indicate SEM. p 0.05; p 0.01; p 0.001.

Article Snippet: Recombinant human BDNF (rhBDNF) was kindly provided by Amgen (Thousand Oaks, CA), recombinant human neurotrophin-4 (NT-4) was generously provided by Genentech (South San Francisco, CA), and anti-rhBDNF neutralizing antibody (mouse IgG1) was obtained from R & D Systems (Minneapolis, MN).

Techniques: Concentration Assay, Control

Journal: The Journal of Neuroscience

Article Title: Local and Target-Derived Brain-Derived Neurotrophic Factor Exert Opposing Effects on the Dendritic Arborization of Retinal Ganglion CellsIn Vivo

doi: 10.1523/jneurosci.22-17-07639.2002

Figure Lengend Snippet: Figure 5. RGC dendritic arborization is temporally sensitive to increased reti- nal BDNF levels. To determine whether RGCs were sensitive to enhanced retinal BDNF in a stage-dependent manner, control or BDNF-treated microspheres were injected into Xenopus retinas at stage 38 or 42. A, The morphology of RGC dendritic arbors revealed a stage- dependent response to increased retinal BDNF levels. B–C, Quantitative analy- sis of dendritic differentiation indicates that earlier exposure to exogenous BDNF (stages 38–45) inhibited den- dritic arborization more dramatically than later exposure to BDNF (stages 42–45). Primary dendrite number as well as dendritic branching was signifi- cantly decreased by altering retinal BDNF starting at stage 38 (B), whereas altering retinal BDNF levels from stage 42 onward (C) selectively reduced den- dritic branching without affecting pri- mary dendrite number. Error bars indi- cate SEM. p 0.05; p 0.001. Scale bar, 10 m.

Article Snippet: Recombinant human BDNF (rhBDNF) was kindly provided by Amgen (Thousand Oaks, CA), recombinant human neurotrophin-4 (NT-4) was generously provided by Genentech (South San Francisco, CA), and anti-rhBDNF neutralizing antibody (mouse IgG1) was obtained from R & D Systems (Minneapolis, MN).

Techniques: Control, Injection

Journal: The Journal of Neuroscience

Article Title: Local and Target-Derived Brain-Derived Neurotrophic Factor Exert Opposing Effects on the Dendritic Arborization of Retinal Ganglion CellsIn Vivo

doi: 10.1523/jneurosci.22-17-07639.2002

Figure Lengend Snippet: Figure 6. RGC axon arbor complexity is unaffected by retinal BDNF levels. To deter- mine whether retinal BDNF influences RGC axon arborization at a distance, tadpoles were intraocularly injected with control, BDNF-, or anti-BDNF-treated microspheres, and the re- sulting changes in RGC axon arbor dynamics were compared with tectally applied BDNF (Cohen-Cory and Fraser, 1995; Lom and Co- hen-Cory, 1999). A, Individual RGC axon ar- bor morphologies of control, retinal BDNF, and tectal BDNF at 0 and 24 hr after treat- ment demonstrate that only tectally applied BDNF significantly alters RGC axon ar- borization. B, C, Altering retinal BDNF lev- els had no significant effects on RGC axon arbor complexity as measured by the in- crease in total branch number (B) and total arbor length (C) 24 and 48 hr after treatment ( p 0.05). Error bars indicate SEM. Scale bar, 20 m.

Article Snippet: Recombinant human BDNF (rhBDNF) was kindly provided by Amgen (Thousand Oaks, CA), recombinant human neurotrophin-4 (NT-4) was generously provided by Genentech (South San Francisco, CA), and anti-rhBDNF neutralizing antibody (mouse IgG1) was obtained from R & D Systems (Minneapolis, MN).

Techniques: Injection, Control

Journal: Journal of neuroscience research

Article Title: Treadmill exercise promotes retinal astrocyte plasticity and protects against retinal degeneration in a mouse model of light-induced retinal degeneration

doi: 10.1002/jnr.25063

Figure Lengend Snippet: Antibody information sheet

Article Snippet: BDNF antibody , Mouse , Monoclonal , Recombinant human BDNF (rhBDNF) expressed in Sf 21 insect cells , Novus Biologicals, NB120-10505 , 1:100 , RRID:AB_788229.

Techniques: Concentration Assay, Residue, Recombinant

Journal: Journal of neuroscience research

Article Title: Treadmill exercise promotes retinal astrocyte plasticity and protects against retinal degeneration in a mouse model of light-induced retinal degeneration

doi: 10.1002/jnr.25063

Figure Lengend Snippet: Probes used in ddPCR Analysis of BDNF and TrkB expression

Article Snippet: BDNF antibody , Mouse , Monoclonal , Recombinant human BDNF (rhBDNF) expressed in Sf 21 insect cells , Novus Biologicals, NB120-10505 , 1:100 , RRID:AB_788229.

Techniques:

Journal: Journal of neuroscience research

Article Title: Treadmill exercise promotes retinal astrocyte plasticity and protects against retinal degeneration in a mouse model of light-induced retinal degeneration

doi: 10.1002/jnr.25063

Figure Lengend Snippet: Increased retinal BDNF–astrocyte interaction observed in treadmill exercised mice. Proximity ligase assay (PLA) was performed on retinal flat mounts from inactive (a and c) and active (b and d) dim and LIRD exposed mice using antibodies targeting BDNF and glutamate transporter-1 (GLT1). GLT1 is an extracellular membrane bound transporter expressed in astrocytes and endothelial cells. Positive co-labeling for astrocytes was done using GFAP (red). Green fluorescence indicates BDNF-GLT1 interaction (expressed in endothelial cells), yellow fluorescence (overlap of green fluorescence from PLA and red fluorescence from GFAP labeling) signifies co-labeling of retinal astrocytes and BDNF. Quantification of PLA fluorescence (green fluorescence overlapping with red fluorescence) revealed a significant increase in BDNF-GLT1 interaction in retinal astrocytes from active groups compared to inactive groups (e). ****p < .0001, scale bars = 15 μm, white box signifies magnified region of image outlined in blue box. N = 4 animals per group, each symbol in the plot represents the average of three biological replicates per retinal quadrant. Experimenter was blinded to experimental conditions and animal group IDs. Two-way ANOVA with Tukey’s multiple comparison analysis was performed. Values are mean ± SD

Article Snippet: BDNF antibody , Mouse , Monoclonal , Recombinant human BDNF (rhBDNF) expressed in Sf 21 insect cells , Novus Biologicals, NB120-10505 , 1:100 , RRID:AB_788229.

Techniques: Membrane, Labeling, Fluorescence, Comparison

Journal: Journal of neuroscience research

Article Title: Treadmill exercise promotes retinal astrocyte plasticity and protects against retinal degeneration in a mouse model of light-induced retinal degeneration

doi: 10.1002/jnr.25063

Figure Lengend Snippet: Magnetic-activated cell sorting reveals exercise alters BDNF and specific TrkB isoform expression in isolated retinal astrocytes. Isolated retinal astrocytes from experimental groups were probed for BDNF (a) and its high-affinity receptor, TrkB, in its catalytically active (TrkB.FL, b) and truncated (TrkB.T1, c) isoforms. Inactive + LIRD retinal astrocytes showed a significant decrease in BDNF expression and a significant increase in TrkB.T1 expression, which has been associated with neuronal cell death. Active + LIRD retinal astrocytes have BDNF expression similar to dim treated groups and had a significant increase in TrkB.FL, which has been associated with cell survival. Bar graphs represent mean values with each data point representing n = 4, n = 12 total per group. Two-way ANOVA with Tukey’s multiple comparison analysis was performed. *p < .05, **p < .01, values are mean ± SD

Article Snippet: BDNF antibody , Mouse , Monoclonal , Recombinant human BDNF (rhBDNF) expressed in Sf 21 insect cells , Novus Biologicals, NB120-10505 , 1:100 , RRID:AB_788229.

Techniques: FACS, Expressing, Isolation, Comparison

Journal: Journal of Nanobiotechnology

Article Title: Novel design of (PEG-ylated)PAMAM-based nanoparticles for sustained delivery of BDNF to neurotoxin-injured differentiated neuroblastoma cells

doi: 10.1186/s12951-020-00673-8

Figure Lengend Snippet: Schematic drawing of PEG-ylated BDNF-PAMAM dendrimer nanoparticles synthesis process as well as BDNF delivery to differentiated 6-OHDA treated neuroblastoma SH-SY5Y cells. Inserted frame comprises molecular structures of BDNF protein, PAMAM and PEG polymer molecules with their overall charge at pH 7.4

Article Snippet: Filtered (centrifree ultrafiltration device, Merck Group, Darmstadt, Germany) stock solutions of carrier free recombinant human BDNF (248-N4-250/CF, R&D Systems, Canada) of known concentrations (typically 250 mgL −1 ) in the phosphate buffered saline (PBS) pH 7.4 ± 0.2, 0.15 M (Biomed, Lublin, Poland) were prepared to remove aggregates and provide constant, free form protein molecules concentration in the solvent.

Techniques: Polymer

Journal: Journal of Nanobiotechnology

Article Title: Novel design of (PEG-ylated)PAMAM-based nanoparticles for sustained delivery of BDNF to neurotoxin-injured differentiated neuroblastoma cells

doi: 10.1186/s12951-020-00673-8

Figure Lengend Snippet: Typical size distribution of BDNF-PAMAM ( a ) and PEG-ylated BDNF-PAMAM ( b ) nanoparticles measured in the bulk by DLS (0.15 M PBS, pH 7.4) without an ultrafiltration process. All values are representative of 5 independent experiments and are expressed as mean ± SD

Article Snippet: Filtered (centrifree ultrafiltration device, Merck Group, Darmstadt, Germany) stock solutions of carrier free recombinant human BDNF (248-N4-250/CF, R&D Systems, Canada) of known concentrations (typically 250 mgL −1 ) in the phosphate buffered saline (PBS) pH 7.4 ± 0.2, 0.15 M (Biomed, Lublin, Poland) were prepared to remove aggregates and provide constant, free form protein molecules concentration in the solvent.

Techniques:

Journal: Journal of Nanobiotechnology

Article Title: Novel design of (PEG-ylated)PAMAM-based nanoparticles for sustained delivery of BDNF to neurotoxin-injured differentiated neuroblastoma cells

doi: 10.1186/s12951-020-00673-8

Figure Lengend Snippet: AFM analysis of PAMAM-BDNF (above part) and PEG-ylated PAMAM-BDNF (below part) nanoparticles adsorbed at mica surface at 0.15 M pH 7.4: PBS. a PAMAM-based nanoparticles at a scan area of 0.5 × 05 µm. b Structure of PAMAM-based nanoparticles after cross-section. c Histogram of adsorbed nanoparticles indicated by direct AFM enumeration, obtained for a low surface molecules concentration. The figure was created by taking into account 10 randomly chosen areas, where each micrograph of PAMAM-based monolayer at the mica surface has a size of 0.5 × 05 µm

Article Snippet: Filtered (centrifree ultrafiltration device, Merck Group, Darmstadt, Germany) stock solutions of carrier free recombinant human BDNF (248-N4-250/CF, R&D Systems, Canada) of known concentrations (typically 250 mgL −1 ) in the phosphate buffered saline (PBS) pH 7.4 ± 0.2, 0.15 M (Biomed, Lublin, Poland) were prepared to remove aggregates and provide constant, free form protein molecules concentration in the solvent.

Techniques: Concentration Assay

Journal: Journal of Nanobiotechnology

Article Title: Novel design of (PEG-ylated)PAMAM-based nanoparticles for sustained delivery of BDNF to neurotoxin-injured differentiated neuroblastoma cells

doi: 10.1186/s12951-020-00673-8

Figure Lengend Snippet: Desorption characteristics of BDNF from PAMAM G5.5 dendrimers-based nanoparticles in PBS electrolyte with increasing loading of protein concentrations from 0.02 to 1 mgL −1 . BDNF detection by ELISA over 24 h incubation of ( a ) PAMAM-BDNF and ( b ) PEG-ylated PAMAM-BDNF nanoparticles

Article Snippet: Filtered (centrifree ultrafiltration device, Merck Group, Darmstadt, Germany) stock solutions of carrier free recombinant human BDNF (248-N4-250/CF, R&D Systems, Canada) of known concentrations (typically 250 mgL −1 ) in the phosphate buffered saline (PBS) pH 7.4 ± 0.2, 0.15 M (Biomed, Lublin, Poland) were prepared to remove aggregates and provide constant, free form protein molecules concentration in the solvent.

Techniques: Enzyme-linked Immunosorbent Assay, Incubation

Journal: Journal of Nanobiotechnology

Article Title: Novel design of (PEG-ylated)PAMAM-based nanoparticles for sustained delivery of BDNF to neurotoxin-injured differentiated neuroblastoma cells

doi: 10.1186/s12951-020-00673-8

Figure Lengend Snippet: Cytotoxicity curves for BDNF, PAMAM-BDNF and PEG-ylated PAMAM-BDNF nanoparticles in differentiated human neuroblastoma cell line SH-SY5Y treated with the 100 µmol/L 6-OHDA neurotoxin. The control (100% viability) are cells treated with 100 µmol/L 6-OHDA only. The data represent means ± SD for 30 experiments

Article Snippet: Filtered (centrifree ultrafiltration device, Merck Group, Darmstadt, Germany) stock solutions of carrier free recombinant human BDNF (248-N4-250/CF, R&D Systems, Canada) of known concentrations (typically 250 mgL −1 ) in the phosphate buffered saline (PBS) pH 7.4 ± 0.2, 0.15 M (Biomed, Lublin, Poland) were prepared to remove aggregates and provide constant, free form protein molecules concentration in the solvent.

Techniques: Control

Journal: Journal of Nanobiotechnology

Article Title: Novel design of (PEG-ylated)PAMAM-based nanoparticles for sustained delivery of BDNF to neurotoxin-injured differentiated neuroblastoma cells

doi: 10.1186/s12951-020-00673-8

Figure Lengend Snippet: Cellular localisation of PAMAM-based nanoparticles in SH-SY5Y cells exposed to 6-OHDA. The cells were incubated either with BDNF-PAMAM-AF488 ( a ) or BDNF-PAMAM-AF488-PEG ( b ) nanoparticles (green color) for different time (5, 10, 30 min, 1 and 24 h) and then co-stained with WGA-Texas Red-X (red) and DAPI (blue). Confocal fluorescence images on the right panel are render series of z-stack with applied surface mode. Panel on the left presents single stack from the z-stack and only nanoparticles (green) and surface glycoproteins (red) are shown. The images were taken at 400 × magnification

Article Snippet: Filtered (centrifree ultrafiltration device, Merck Group, Darmstadt, Germany) stock solutions of carrier free recombinant human BDNF (248-N4-250/CF, R&D Systems, Canada) of known concentrations (typically 250 mgL −1 ) in the phosphate buffered saline (PBS) pH 7.4 ± 0.2, 0.15 M (Biomed, Lublin, Poland) were prepared to remove aggregates and provide constant, free form protein molecules concentration in the solvent.

Techniques: Incubation, Staining, Fluorescence

Journal: Journal of Nanobiotechnology

Article Title: Novel design of (PEG-ylated)PAMAM-based nanoparticles for sustained delivery of BDNF to neurotoxin-injured differentiated neuroblastoma cells

doi: 10.1186/s12951-020-00673-8

Figure Lengend Snippet: Desorption characteristic of BDNF from PAMAM G5.5 dendrimers-based nanoparticles for cell culture supernatant under increasing nanocarriers loading of protein concentrations from 0.02 to 2 mgL −1 . BDNF detection by ELISA over 24 h in cells incubated with BDNF, PAMAM-BDNF nanoparticles and PEG-ylated PAMAM-BDNF nanoparticles. The data represent means ± SD for five experiments

Article Snippet: Filtered (centrifree ultrafiltration device, Merck Group, Darmstadt, Germany) stock solutions of carrier free recombinant human BDNF (248-N4-250/CF, R&D Systems, Canada) of known concentrations (typically 250 mgL −1 ) in the phosphate buffered saline (PBS) pH 7.4 ± 0.2, 0.15 M (Biomed, Lublin, Poland) were prepared to remove aggregates and provide constant, free form protein molecules concentration in the solvent.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Incubation

Journal: Journal of Diabetes Research

Article Title: Plasma BDNF Is Reduced in Acute Ischemic Stroke Patients With Type 2 Diabetes Mellitus and Associated With Fibrin-Rich Thrombi

doi: 10.1155/jdr/6693363

Figure Lengend Snippet: BDNF lower in AIS/DM patients. (a) Expression of plasma BDNF in healthy controls, AIS/non-DM patients, and AIS/DM patients. (b) Representative images of immunohistochemical detection of BDNF (brown) in clots from AIS/non-DM and AIS/DM patients. (c) The column graph represents mean ± SEM of staining scores in each group. (d) Representative clots from AIS/non-DM and AIS/DM patients were stained using MSB to visualize the fibrin (dark pink to red), RBCs (yellow), and platelets (gray). (e) Column graph displays the differences in fibrin content in clots. (f) Fibrin fluorescence of clots from AIS patients. (g) The bar graph represents mean ± SEM fluorescence values.

Article Snippet: Citrated plasma aliquots (100 μ L), underwent 15-min preincubation at 37°C with either (a) 1- μ L distilled water containing graded recombinant human BDNF (rHu-BDNF) qualities (0.1, 0.2, or 0.3 ng, Solarbio P00062) or (b) equivalent-volume distilled water (control).

Techniques: Expressing, Clinical Proteomics, Immunohistochemical staining, Staining, Fluorescence

Journal: Journal of Diabetes Research

Article Title: Plasma BDNF Is Reduced in Acute Ischemic Stroke Patients With Type 2 Diabetes Mellitus and Associated With Fibrin-Rich Thrombi

doi: 10.1155/jdr/6693363

Figure Lengend Snippet: The clot permeability and ultrastructure. (a) Thrombus permeability was significantly lower in AIS/DM patients than AIS patients without DM. (b) Correlation between BDNF concentrations and thrombus permeability. (c) BDNF levels in patients based on the severity of stroke. (d) Scanning electron micrographs of thrombi. Arrows indicate fibrin; dotted lines indicate fibrin band. Bar = 20 μ m. (e) Transmission electron micrographs of thrombi from the AIS/non-DM and AIS/DM patients. Arrows indicate fibrin; dotted lines indicate fibrin band. Bar = 5 μ m.

Article Snippet: Citrated plasma aliquots (100 μ L), underwent 15-min preincubation at 37°C with either (a) 1- μ L distilled water containing graded recombinant human BDNF (rHu-BDNF) qualities (0.1, 0.2, or 0.3 ng, Solarbio P00062) or (b) equivalent-volume distilled water (control).

Techniques: Permeability, Transmission Assay

Journal: Journal of Diabetes Research

Article Title: Plasma BDNF Is Reduced in Acute Ischemic Stroke Patients With Type 2 Diabetes Mellitus and Associated With Fibrin-Rich Thrombi

doi: 10.1155/jdr/6693363

Figure Lengend Snippet: BDNF influences fibrin density. (a) A ribbon diagram illustrates the binding of BDNF to the heparin-binding domain of fibrin(ogen), with the structure generated via DMFold. (b) rHu-BDNF was added to plasma pools from AIS/DM patients before induction of coagulation with thrombin; then, the fibrin density of the clot was analyzed. Visualization images of clots using rabbit polyclonal fibrinogen/FITC (bar = 100 μ m) and (c) the percentage of fibrin fibers versus control. Fibrin fibers were analyzed using ImageJ software. All samples were performed in triplicate. Data are expressed as mean ± SEM, ⁣ ∗ p < 0.05.

Article Snippet: Citrated plasma aliquots (100 μ L), underwent 15-min preincubation at 37°C with either (a) 1- μ L distilled water containing graded recombinant human BDNF (rHu-BDNF) qualities (0.1, 0.2, or 0.3 ng, Solarbio P00062) or (b) equivalent-volume distilled water (control).

Techniques: Binding Assay, Generated, Clinical Proteomics, Coagulation, Control, Software

Journal: Journal of Diabetes Research

Article Title: Plasma BDNF Is Reduced in Acute Ischemic Stroke Patients With Type 2 Diabetes Mellitus and Associated With Fibrin-Rich Thrombi

doi: 10.1155/jdr/6693363

Figure Lengend Snippet: Observing the fibrin diameter by scanning electron microscopy. (a) Fibrin structures formed from venous blood from AIS/DM subjects were added with different concentrations of BDNF. The ultrastructure of fibrin was observed by a scanning electron microscope. (b) The column chart shows the difference in the diameter of fibrin fibers in clots of different groups. ⁣ ∗ p < 0.05 and ⁣ ∗∗∗ p < 0.001.

Article Snippet: Citrated plasma aliquots (100 μ L), underwent 15-min preincubation at 37°C with either (a) 1- μ L distilled water containing graded recombinant human BDNF (rHu-BDNF) qualities (0.1, 0.2, or 0.3 ng, Solarbio P00062) or (b) equivalent-volume distilled water (control).

Techniques: Electron Microscopy, Microscopy

Journal: Journal of Diabetes Research

Article Title: Plasma BDNF Is Reduced in Acute Ischemic Stroke Patients With Type 2 Diabetes Mellitus and Associated With Fibrin-Rich Thrombi

doi: 10.1155/jdr/6693363

Figure Lengend Snippet: rHu-BDNF influences fibrin polymerization, coagulation, and lysis. (a) Representative turbidity curves were recorded spectrophotometrically at 350 nm (37°C) with 30-s intervals. (b) Maximum turbidity values were determined under identical monitoring conditions. (c) Maximum clot firmness was assessed by thromboelastographic analyses (MA). Plasma samples supplemented with recombinant BDNF prior to thrombin/tPA-induced coagulation underwent subsequent polymerization analysis. (d) Thrombin clotting time (TT) and (e) lysis time were measured. All samples were performed in triplicate. Data are expressed as mean ± SEM, n = 3 different pools. ⁣ ∗ p < 0.05, ⁣ ∗∗ p < 0.01, and ⁣ ∗∗∗ p < 0.001.

Article Snippet: Citrated plasma aliquots (100 μ L), underwent 15-min preincubation at 37°C with either (a) 1- μ L distilled water containing graded recombinant human BDNF (rHu-BDNF) qualities (0.1, 0.2, or 0.3 ng, Solarbio P00062) or (b) equivalent-volume distilled water (control).

Techniques: Coagulation, Lysis, Clinical Proteomics, Recombinant

Journal: bioRxiv

Article Title: Dynamic changes in chromosome and nuclear architecture during maturation of normal and ALS C9orf72 motor neurons

doi: 10.1101/2025.09.22.677835

Figure Lengend Snippet: a) Brightfield images of three different cell-types (scale bar=100 μm). b) Schematic view of neural differentiation from IPSCs into motor neuron and motor neuron maturation. IPSCs are treated with listed small molecules for 14 days. At day 14, cells were dissociated and sorted. NCAM+ and EPCAM-cells (population highlighted in red) were collected and plated as immature post-mitotic neurons that were cultured 6 more weeks for maturation (SB-431542: TGF-beta/Smad inhibitor; LDN-193189: BMP pathway inhibitor; RA: retinoic acid; SAG: Smoothened agonist; DAPT: γ-secretase inhibitor; SU-5402: FGFR1 inhibitor; BDNF: brain-derived neurotrophic factor; GDNF: glial-derived neurotrophic factor; CNTF: ciliary neurotrophic factor. c) Schematic view of the study design: primary fibroblasts of two healthy controls and two ALS patients with C9orf72 mutation were reprogrammed into IPSCs. Two independent IPSC clones for each line were picked for neural differentiations. Hi-C and RNA-seq libraries were prepared for each clone. d) TPM counts of PF related genes were shown for PF (red), IPSC (salmon) and MN (blue) cell-types. e) TPM counts of IPSC related genes were shown for PF, IPSC and MN cell-types. f) TPM counts of MN and glial related genes were shown for PF, IPSC and MN cell-types.

Article Snippet: The sorted EpCAM-negative NCAM-positive cells were collected and plated onto Poly-L-lysine coated plates (Sigma, P5899) with motor neuron media (Neurobasal medium, 1x N-2 supplement, 1x B-27 supplement, 1x GlutaMax and 1x MEM NEAA solution) with 10 μM ROCK inhibitor and 10 ng per ml of the following neurotrophic factors: glial cell-derived neurotrophic factor (GDNF) (R&D systems, cat. no. 212-GD-010/CF), brain-derived neurotrophic factor (BDNF) (R&D systems, cat. no. 11166-BD-010) and ciliary neurotrophic factor (CNTF) (R&D systems, cat. no. 257-NT-010/CF).

Techniques: Cell Culture, Derivative Assay, Mutagenesis, Clone Assay, Hi-C, RNA Sequencing