human probdnf  (Biosensis ltd)

Journal: Science Advances

Article Title: BDNF-driven synaptic plasticity requires autocrine matrix metalloproteinase–9 activity

doi: 10.1126/sciadv.adx2369

Figure Lengend Snippet: ( A ) Scheme of the TrkB FRET sensor. Created in BioRender. Kalita, K. (2025) https://BioRender.com/qaducze . ( B ) Example FLIM images showing TrkB activation. Warmer colors represent higher TrkB activity. Yellow cross indicates uncaging spot. Scale bar, 1 μm. ( C ) Averaged TrkB activation changes (Δ binding fraction) in dendritic spines following uncaging in the presence of DMSO or Inhibitor I. Data are means ± SEM. Gray box indicates uncaging period. ( D ) Statistical analysis of (C). Averaged TrkB activation in stimulated spines during transient (1 to 3 min) and sustained phase (9 to 11 min). Gray dots, individual spines; bars, means ± SEM. DMSO (blue; n = 70 spines, 27 cells, 16 animals) and Inhibitor I (red; n = 49 spines, 21 cells, 10 animals). Repeated-measures ANOVA: Time ( P = 0.0010); Inhibitor ( P = 0.0013); Time × Inhibitor ( P = 0.7403), followed by Šídák’s multiple comparisons test ( P values indicated on the graph). ( E ) Averaged TrkB activation changes in dendritic spines following uncaging in WT or MMP-9 KO slices. All markings as in (C). ( F ) Statistical analysis of (E). All markings as in (D). WT (blue, n = 66 spines; 22 cells, 10 animals), MMP-9 KO (yellow, n = 73 spines; 25 cells, 11 animals). Repeated-measures ANOVA: Time ( P = 0.0257); MMP-9 KO ( P = 0.0054); Time × MMP-9 KO ( P = 0.7238), followed by Šidák’s multiple comparison test ( P values indicated on the graph). ( G ) Example immunoblot of digestion reaction of proBDNF incubated with either active MMP-9, inactive MMP-9 (E402A), or the reaction buffer. Bands correspond to proBDNF (~26 kDa) and mBDNF (~14 kDa). ( H ) Quantification of immunoblots of three digestion reactions. Gray dots, individual values of mBDNF band intensity in separate experiments; bars, mean ± SEM. One-way ANOVA ( P = 0.0021) followed by Tukey’s multiple comparisons test ( P values indicated on the graph).

Article Snippet: Twenty nanograms of recombinant proBDNF (Alomone Labs) was incubated with 50 ng of recombinant MMP-9 (Calbiochem) or 50 ng of recombinant, human, inactive MMP-9 (E402A) in total volume of 20 μl.

Techniques: Activation Assay, Activity Assay, Binding Assay, Comparison, Western Blot, Incubation

Journal: Science Advances

Article Title: BDNF-driven synaptic plasticity requires autocrine matrix metalloproteinase–9 activity

doi: 10.1126/sciadv.adx2369

Figure Lengend Snippet: Activation of NMDAR (1) leads to the release of MMP-9 and BDNF (2), which might be released in its either pro-form or mature form with a propeptide. (3) tPA activates plasminogen to plasmin, which can also activate proMMP-9. (4) Plasmin and MMP-9 can extracellularly process proBDNF to mBDNF, which activates its receptor—TrkB (5). TrkB activation, together with other intracellular signaling, leads to the LTP cascade, including actin polymerization and cytoskeleton remodeling causing spine enlargement. (6) ProBDNF and BDNF propeptide, which is also co-released with mBDNF, can activate p75 TNR , leading to LTD. (7) It is possible that MMP-9 can reduce bioactive BDNF propeptide and promote a competing TrkB activation. (8) Both plasmin and MMP-9 are blocked by their inhibitors, which control their action. Created in BioRender. Kalita, K. (2025) https://BioRender.com/nbak5x7 .

Article Snippet: Twenty nanograms of recombinant proBDNF (Alomone Labs) was incubated with 50 ng of recombinant MMP-9 (Calbiochem) or 50 ng of recombinant, human, inactive MMP-9 (E402A) in total volume of 20 μl.

Techniques: Activation Assay, Control

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