Journal: bioRxiv

Article Title: CNTF specifically slows down the axonal transport of signalling endosomes

doi: 10.1101/2025.10.09.681259

Figure Lengend Snippet: A) Male and female ChAT.eGFP mice were allocated into one of five groups: i) PBS (control); ii) CNTF; iii) proBDNF; iv) HGF; and v) NTRN. B) Each mouse received intramuscular injections of H C T combined with a single NTF into the left tibialis anterior and right soleus muscles to target fast and slow motor neurons, respectively. After a 4-8 h incubation period, time-lapse microscopy was performed on both sciatic nerves. C) H C T-labelled signalling endosomes (pseudo-coloured in magenta) from single ChAT.eGFP motor axons were individually tracked to quantify retrograde transport dynamics. Three representative retrogradely transported signalling endosomes are identified by yellow, cyan, and peach arrowheads connected by dashed lines across frames. Grey arrowheads and dashed lines identify a stationary endosome. See also Video 1 . Scale bar = 5 μm, frame interval = 3 s.

Article Snippet: Mice were divided into five experimental cohorts, and received 5-7 μg of H C T mixed with: a) phosphate buffered saline (PBS) as the control; b) 50 ng recombinant human CNTF protein (Peprotech, 450-13), c) 50 ng recombinant mouse cleavage-resistant proBDNF (Alomone labs, B-243), d) 25 ng recombinant human HGF (Bio-Techne, 294-HG[CF]), or e) 50 ng recombinant human NTRN (Peprotech, 450-11), as summarised in .

Techniques: Control, Muscles, Incubation, Time-lapse Microscopy

Journal: bioRxiv

Article Title: CNTF specifically slows down the axonal transport of signalling endosomes

doi: 10.1101/2025.10.09.681259

Figure Lengend Snippet: In both fast motor neurons (FMNs) and slow motor neurons (SMNs), proBDNF did not alter A) mean endosome speed ( p = 0.06 for motor neuron type; p = 0.19 for stimulation factor; p = 0.70 for interaction), B) maximum endosome speed ( p = 0.30 for motor neuron type; p = 0.62 for stimulation factor; p = 0.25 for interaction) or C) pausing percentage ( p = 0.24 for motor neuron type; p = 0.92 for stimulation factor; p = 0.34 for interaction). D ) Violin plots of individual endosomes show comparable distributions in all conditions (FMNs: PBS mean = 2.78 µm/s ± 0.03 [n = 495], proBDNF mean = 2.67 µm/s ± 0.03 [n = 496]; SMNs: PBS mean = 2.57 µm/ s ± 0.03 [n = 495], proBDNF mean = 2.53 µm/s ± 0.03 [n = 477]). Overlapping endosome frame-to-frame and mean endosome speed distribution curves confirm that proBDNF does not modulate retrograde transport in E) FMNs or F) SMNs. Statistical analyses were performed using two-way ANOVA and Holm-Šídák 1 s multiple comparisons tests. ns, not significant. n = 7-8. Black circles = males (n = 4 PBS; n = 2 CNTF), white circles = females (n = 4 PBS; n = 5 proBDNF).

Article Snippet: Mice were divided into five experimental cohorts, and received 5-7 μg of H C T mixed with: a) phosphate buffered saline (PBS) as the control; b) 50 ng recombinant human CNTF protein (Peprotech, 450-13), c) 50 ng recombinant mouse cleavage-resistant proBDNF (Alomone labs, B-243), d) 25 ng recombinant human HGF (Bio-Techne, 294-HG[CF]), or e) 50 ng recombinant human NTRN (Peprotech, 450-11), as summarised in .

Techniques:

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