Journal: Scientific Reports

Article Title: Peripheral Brain Derived Neurotrophic Factor Precursor Regulates Pain as an Inflammatory Mediator

doi: 10.1038/srep27171

Figure Lengend Snippet: ( A ) Representative Western blot (a) and their semi-quantitative analyses of mature BDNF (b), proBDNF (c) and their ratio (d) in the local tissue after 10 μL 5% formalin intra-plantar injection into Kunming mice (*p < 0.05, **p < 0.01 versus control, one-way ANOVA followed by Dunnett’s Multiple Comparison post hoc test, n = 3 per group). ( B ) H&E staining (a and d) and immunohistochemsitry (b,c and e–g) of proBDNF in the foot skin at 3 h post-formalin injection. proBDNF is expressed in the epidermis, basal layer and subcutaneous layers in the foot skin (b,c); Higher magnification (box in b) showing proBDNF is also mildly expressed in the nerve fibers in the control plantar (c); Responding to peripheral inflammation by 5% formalin intra-plantar injection, intensive proBDNF immunoreactivity is observed and mainly localized in the inflammatory cells (f, black arrows) and nerve fiber-like structures (g). Scale bars: 50 μm, 3 replicates, n = 3 per group. ( C ) a, Representative Western blot of proBDNF and mBDNF; b–d, Semi-quantitative analyses of mBDNF, proBDNF and their ratio in the inflamed tissue after Complete Freund Adjuvant (CFA, 10 μL) intra-plantar injection into Kunming mice (*p < 0.05, **p < 0.01versus control, one-way ANOVA followed by Dunnett’s Multiple Comparison post hoc test, n = 3 per group). ( D ) Histological staining (a) and proBDNF immunohistochemistry (b,c) in the plantar at 1 day post-CFA injection; c, higher magnification of box in b showing that proBDNF is highly expressed in the inflammatory cells. Scale bar, 100 μm, 3 replicates, n = 3 per group. Data bars represent mean ± s.e.m.

Article Snippet: For ELISA assay, 96-well polystyrene microtiter plates (Nunc, Roskilde, Denmark, USA) were coated with the generated human proBDNF prodomain, and commercial human, rat and mice proBDNF proteins (Alomone Labs, Israel), and human mBDNF (Alomone Labs, Israel) (all at 1 μg/mL with total volume 50 μL) overnight at 4 °C.

Techniques: Western Blot, Injection, Control, Comparison, Staining, Adjuvant, Immunohistochemistry

Journal: Scientific Reports

Article Title: Peripheral Brain Derived Neurotrophic Factor Precursor Regulates Pain as an Inflammatory Mediator

doi: 10.1038/srep27171

Figure Lengend Snippet: ( A ) Dosage effect of exogenous proBDNF protein on PWT by injection of proBDNF protein into the plantar (*P < 0.05, **p < 0.01 versus baseline, one-way ANOVA followed by Dunnett’s Multiple Comparison post hoc test, n = 10–12 per group). ( B ) Ectopic overexpression of proBDNF by intra-plantar injection of Ad-proBDNF or Ad-EGFP reduces PWT dramatically in Kunming mice. (a) Representative proBDNF Western blot (upper panel) and its semi-quantitative analysis (lower panel, *P < 0.05, ***p < 0.001 versus control, ### p < 0.001 versus indicated groups, one-way ANOVA followed by Tukey’s Multiple Comparison post hoc test, n = 4 per group); (b) Representative fluorescent images after delivery of Ad-EGFP . Scale bar, 50 μm, 3 replicates, n = 3 per group; (c) PWT at 7 days post-injection of Ad-proBDNF or Ad-EGFP control (***p < 0.001 versus Ad-EGFP , student’s t test, n = 7 per group). ( C ) Co-injection of proBDNF (0.1 μg), but not mBDNF (0.1 μg) restored the biphasic nociceptive response after low-concentration of formalin (0.5%) intra-plantar injection (*p < 0.05versus vehicle, two-way ANOVA followed by Bonferroni’s Multiple Comparison post hoc test, n = 10–12 per group). ( D ) Exogenous proBDNF (1 μg) intra-plantar injection induces ERK activation in the ipsilateral spinal cord dorsal horn at 3 h post-injection, Scale bar, 50 μm, n = 3 per group, 3 replicates. ( E ) Spinal p-ERK expression at 3 h after proBDNF (1 μg) intra-plantar injection. (a) Representative Western blot and (b,c) their semi-quantitative analyses of p-ERK, (*p < 0.05, **p < 0.01 versus proBDNF-R, one-way ANOVA followed by Dunnett’s Multiple Comparison post hoc test, n = 3 per group). Data bars represent mean ± s.e.m.

Article Snippet: For ELISA assay, 96-well polystyrene microtiter plates (Nunc, Roskilde, Denmark, USA) were coated with the generated human proBDNF prodomain, and commercial human, rat and mice proBDNF proteins (Alomone Labs, Israel), and human mBDNF (Alomone Labs, Israel) (all at 1 μg/mL with total volume 50 μL) overnight at 4 °C.

Techniques: Injection, Comparison, Over Expression, Western Blot, Control, Concentration Assay, Activation Assay, Expressing

Journal: Scientific Reports

Article Title: Peripheral Brain Derived Neurotrophic Factor Precursor Regulates Pain as an Inflammatory Mediator

doi: 10.1038/srep27171

Figure Lengend Snippet: ( A ) ELISA assay for the immunoreactivity of 2B11 against human proBDNF prodomain, and human, rat and mice proBDNF proteins, and human mature BDNF (mBDNF). 2B11 has strong immunoreactivity against proBDNF and prodomain, but not mBDNF; ( B ) Representative Western blot of human proBDNF and mBDNF detected by 2B11 (dilution 1:2000), note that 2B11 specifically recognizes proBDNF, but not mBDNF. ( C ) Representative images of neurosphere radiant migration treated by proBDNF, mBDNF, sheep polyclonal anti-proBDNF antibody, mouse monoclonal anti-proBDNF antibody 2B11 and co-treatment. ( D ) Statistical analysis of neurosphere migration radiance assay (***P < 0.001 versus control, # p < 0.05 versus indicated group, one-way ANOVA followed by Tukey’s Multiple Comparison post hoc test). Neurospheres treated with proBDNF (100 ng/mL) showed dormancy without any neuronal migration and neurospheres had no morphological changes. Neurospheres treated with 2B11 (100 ng/ml) showed strong migration capability comparing with other groups. Neurospheres treated with 2B11 and proBDNF (100 ng/ml) showed similar ability of migration with sheep anti-proBDNF antibody treatment group.

Article Snippet: For ELISA assay, 96-well polystyrene microtiter plates (Nunc, Roskilde, Denmark, USA) were coated with the generated human proBDNF prodomain, and commercial human, rat and mice proBDNF proteins (Alomone Labs, Israel), and human mBDNF (Alomone Labs, Israel) (all at 1 μg/mL with total volume 50 μL) overnight at 4 °C.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Migration, Control, Comparison

Journal: Scientific reports

Article Title: Cognitive and sensorimotor benefits of moderate- and high-intensity exercise are associated with specific expression of neurotrophic markers in older rats.

doi: 10.1038/s41598-025-90719-4

Figure Lengend Snippet: Fig. 1. Experimental protocol. All trained animals carried out 4 weeks of training, 5 times per week either in MICT or HIIT. All animals performed both incremental test and grip strength at PRE, 2W and POST, while they only performed adhesive removal test and the novel object recognition test at the end of the training (in POST). During the incremental test, blood lactate was collected at every level to determine the SLT. At the end of the protocol (48 h after the last incremental test) both hippocampi and cortices were removed to perform Western blot and ELISA measurements.

Article Snippet: Quantification of pro and mature BDNF standard and cortical samples was respectively performed with proBDNF Rapid ELISA Kit (Biosensis®, BEK-2217-2P – sandwich ELISA–Thebarton, SA, Australia) and mBDNF Rapid ELISA Kit Kit (Biosensis®, BEK-2211-1P/2P–sandwich ELISA–Thebarton, SA, Australia) in the concentrated solutions following the manufacturer’s protocol.

Techniques: Adhesive, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Scientific reports

Article Title: Cognitive and sensorimotor benefits of moderate- and high-intensity exercise are associated with specific expression of neurotrophic markers in older rats.

doi: 10.1038/s41598-025-90719-4

Figure Lengend Snippet: Fig. 4. Effect of MICT and HIIT programs on cortical protein levels. (a) Level of cortical mBDNF (n = 19) measured by ELISA (in pg/ml-1) in MICT, HIIT and Control groups at POST training. Level of cortical (b) TrkB (n = 20), (c) pTrkB (n = 20), (d), p757NTR (n = 19), (e) NKCC1/KCC2 ratio (n = 19), (f) KCC2 (n = 18) and (g) CTSB (n = 20) in MICT, HIIT and Control groups at POST training. Protein level measured by Western blot, was calculated as the ratio of the protein of interest divided by the protein of normalization relative to the control. The normalization protein was the α-tubulin for all the molecules except for the KCC2 which was β3-tubulin. Pictures of Western blot membranes are shown below each graph. Dashed lines were added to separate the groups. Note that the α-tubulin used for TrkB (c) was the same used for pTrkB (d), as membranes were stripped between these two quantifications. The samples derived from the same experiment and that gels/ blots were systematically processed in parallel the same day (2 gels per protein to reach a sufficient number of samples). *Significant differences in protein levels between groups. Data is expressed in mean ± SD (a–f) and in median (min to max) (g).

Article Snippet: Quantification of pro and mature BDNF standard and cortical samples was respectively performed with proBDNF Rapid ELISA Kit (Biosensis®, BEK-2217-2P – sandwich ELISA–Thebarton, SA, Australia) and mBDNF Rapid ELISA Kit Kit (Biosensis®, BEK-2211-1P/2P–sandwich ELISA–Thebarton, SA, Australia) in the concentrated solutions following the manufacturer’s protocol.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Western Blot, Derivative Assay

Journal: Scientific reports

Article Title: Cognitive and sensorimotor benefits of moderate- and high-intensity exercise are associated with specific expression of neurotrophic markers in older rats.

doi: 10.1038/s41598-025-90719-4

Figure Lengend Snippet: Fig. 5. Effect of MICT and HIIT programs on hippocampal protein levels. (a) Level of hippocampal IGF- 1(n = 21) measured by ELISA in MICT, HIIT and Control groups at POST training. Level of hippocampal (b) IGFB-2 (n = 21), (c) p75NTR (n = 22), (d) TrkB (n = 21), (e) CTSB (n = 22), (f) NKCC1 (n = 21), (g) KCC2 (n = 20), and (h) the NKCC1/KCC2 ratio (n = 18) measured by Western blot in MICT, HIIT and Control groups at POST. Protein level was calculated as the ratio of the protein of interest divided by the protein of normalization relative to the control. The normalization protein was the α-tubulin for all the molecules except for the KCC2 which was β3-tubulin. Pictures of the Western blot membranes are shown below each graph. Dashed lines were added to separate the groups. Note that the α-tubulin used for p75NTR (c) was the same used for NKCC1 (f) as their molecular weight are different, the quantification of both was conducted on the same membranes. The samples derived from the same experiment and that gels/blots were processed in parallel the same day (2 gels per protein to reach a sufficient number of samples). *Significant differences in protein levels between groups. All data is expressed in mean ± SD.

Article Snippet: Quantification of pro and mature BDNF standard and cortical samples was respectively performed with proBDNF Rapid ELISA Kit (Biosensis®, BEK-2217-2P – sandwich ELISA–Thebarton, SA, Australia) and mBDNF Rapid ELISA Kit Kit (Biosensis®, BEK-2211-1P/2P–sandwich ELISA–Thebarton, SA, Australia) in the concentrated solutions following the manufacturer’s protocol.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Western Blot, Molecular Weight, Derivative Assay

Journal: Journal of Clinical and Translational Research

Article Title: Bacopa monnieri supplementation has no effect on serum brain-derived neurotrophic factor levels but beneficially modulates nuclear factor kappa B and cyclic AMP response element-binding protein levels in healthy elderly subjects

doi:

Figure Lengend Snippet: Average±SD for BDNF levels, MoCA scores and GDS score at each visit

Article Snippet: Pro-BDNF and mBDNF levels were measured by ELISAs using the human proBDNF and the mBDNF ELISAs from Aviscera-Bioscience (CA, USA) which show reproducible, sensitive, and selective detection of both forms of BDNF in human serum [ ].

Techniques: