ant006  (Alomone Labs)

Journal: Frontiers in Cell and Developmental Biology

Article Title: Requirements of Postnatal proBDNF in the Hippocampus for Spatial Memory Consolidation and Neural Function

doi: 10.3389/fcell.2021.678182

Figure Lengend Snippet: Blockage of proBDNF expression during the postnatal period induces spatial learning impairments. (A) The level of proBDNF in the hippocampus. (* p

Article Snippet: A total of 418 male offspring from an average of 84 litters were randomly assigned to one of six groups: ( ) anti-proBDNF (second week), ( ) anti-proBDNF (fourth week), and ( ) anti-proBDNF (eighth week) groups received bilateral infusion of rabbit polyclonal anti-proBDNF antibody ( ; ) in the CA1 region of the HPC throughout the entire second postnatal week (PD2w, from PD8 to PD14), fourth postnatal week (PD4w, from PD22 to PD28), and eighth postnatal week (PD8w, from PD50 to PD56), respectively; ( ) control group was treated with the same volume of the vehicle (artificial cerebrospinal fluid, ACSF) throughout the whole PD2w (Con@2w), PD4w (Con@4w), and PD8w (Con@8w); ( ) Anti+TBOA group, which received infusion of anti-proBDNF antibody during the postnatal weeks, was bilaterally infused with DL-threo-β-benzyloxyaspartate (DL-TBOA) 0.5 or 2.5 h before spatial training [Anti+TBOA0.5(a) or Anti+TBOA2.5(a)], immediately following behavioral training [Anti+TBOA(b)] or 0.5 h before probe test [Anti+TBOA(c)]; and ( ) control group, which received infusion of ACSF during the postnatal weeks, was bilaterally infused with DL-TBOA 0.5 h before spatial training [TBOA(a)], immediately following behavioral training [TBOA(b)] or 0.5 h before probe test [TBOA(c)]; ( ) naive group was reared as the control group without the treatment.

Techniques: Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Requirements of Postnatal proBDNF in the Hippocampus for Spatial Memory Consolidation and Neural Function

doi: 10.3389/fcell.2021.678182

Figure Lengend Snippet: Schematic representations of the cannulae and electrode placements and morphological alterations in the CA1 region. (A) Histological (left) and schematic (right) representations of the cannula placements. The control group infused with ACSF throughout the whole PD4w; the Anti@2w and Anti@4w groups were infused with anti-proBDNF antibody throughout the whole second and fourth postnatal weeks, respectively. The yellow arrows indicated the top of the cannulae. (B) Histological and schematic representations of electrode placements. (C) Following the open field test, infusion-induced neuronal damage was assessed by Silver staining (see Supplementary Methods ). The white scale bar presented at the bottom of the photomicrograph indicated 25 μm. The yellow arrows indicated the electrode tips. There was no statistical difference in the quantification of neurodegeneration in CA1 neurons between the control (top) and anti-proBDNF (bottom) groups. The anti-proBDNF group was infused with anti-proBDNF antibody throughout the whole fourth postnatal week. The control group was treated with the same volume of the vehicle (ACSF) throughout the whole the fourth postnatal week. The treatment was conducted twice a day in a 12-h interval. n = 6 for each group.

Article Snippet: A total of 418 male offspring from an average of 84 litters were randomly assigned to one of six groups: ( ) anti-proBDNF (second week), ( ) anti-proBDNF (fourth week), and ( ) anti-proBDNF (eighth week) groups received bilateral infusion of rabbit polyclonal anti-proBDNF antibody ( ; ) in the CA1 region of the HPC throughout the entire second postnatal week (PD2w, from PD8 to PD14), fourth postnatal week (PD4w, from PD22 to PD28), and eighth postnatal week (PD8w, from PD50 to PD56), respectively; ( ) control group was treated with the same volume of the vehicle (artificial cerebrospinal fluid, ACSF) throughout the whole PD2w (Con@2w), PD4w (Con@4w), and PD8w (Con@8w); ( ) Anti+TBOA group, which received infusion of anti-proBDNF antibody during the postnatal weeks, was bilaterally infused with DL-threo-β-benzyloxyaspartate (DL-TBOA) 0.5 or 2.5 h before spatial training [Anti+TBOA0.5(a) or Anti+TBOA2.5(a)], immediately following behavioral training [Anti+TBOA(b)] or 0.5 h before probe test [Anti+TBOA(c)]; and ( ) control group, which received infusion of ACSF during the postnatal weeks, was bilaterally infused with DL-TBOA 0.5 h before spatial training [TBOA(a)], immediately following behavioral training [TBOA(b)] or 0.5 h before probe test [TBOA(c)]; ( ) naive group was reared as the control group without the treatment.

Techniques: Silver Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: Requirements of Postnatal proBDNF in the Hippocampus for Spatial Memory Consolidation and Neural Function

doi: 10.3389/fcell.2021.678182

Figure Lengend Snippet: Blocking proBDNF reduces spine number and learning-related GluN2B expression. The samples from rats that performed spatial training in the MWM task were collected immediately following the training phase and selected for detecting spine density and the expression of glutamatergic receptor subunits. (A) Spine alteration in naive, untrained-antiproBDNF, trained control, and trained-antiproBDNF rats (top). Quantification of spine density (middle) and the proportion of spine (bottom). Scale bars, 5 μm. n = 6 per group. Sample images were projected at minimal intensity and inverted, background was then subtracted, followed by brightness/contrast adjustment. The expression and phosphorylation of GluA1 (B) and the expression of GluG2/3 and the phosphorylation of GluA2 (C) of AMPAR subunits. n = 10 per group. The expression and phosphorylation of GluN2A (D) and GluN2B (E) of NMDAR subunits. (* p

Article Snippet: A total of 418 male offspring from an average of 84 litters were randomly assigned to one of six groups: ( ) anti-proBDNF (second week), ( ) anti-proBDNF (fourth week), and ( ) anti-proBDNF (eighth week) groups received bilateral infusion of rabbit polyclonal anti-proBDNF antibody ( ; ) in the CA1 region of the HPC throughout the entire second postnatal week (PD2w, from PD8 to PD14), fourth postnatal week (PD4w, from PD22 to PD28), and eighth postnatal week (PD8w, from PD50 to PD56), respectively; ( ) control group was treated with the same volume of the vehicle (artificial cerebrospinal fluid, ACSF) throughout the whole PD2w (Con@2w), PD4w (Con@4w), and PD8w (Con@8w); ( ) Anti+TBOA group, which received infusion of anti-proBDNF antibody during the postnatal weeks, was bilaterally infused with DL-threo-β-benzyloxyaspartate (DL-TBOA) 0.5 or 2.5 h before spatial training [Anti+TBOA0.5(a) or Anti+TBOA2.5(a)], immediately following behavioral training [Anti+TBOA(b)] or 0.5 h before probe test [Anti+TBOA(c)]; and ( ) control group, which received infusion of ACSF during the postnatal weeks, was bilaterally infused with DL-TBOA 0.5 h before spatial training [TBOA(a)], immediately following behavioral training [TBOA(b)] or 0.5 h before probe test [TBOA(c)]; ( ) naive group was reared as the control group without the treatment.

Techniques: Blocking Assay, Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Requirements of Postnatal proBDNF in the Hippocampus for Spatial Memory Consolidation and Neural Function

doi: 10.3389/fcell.2021.678182

Figure Lengend Snippet: Activation of GluN2B can rescue memory consolidation induced by blocking postnatal proBDNF. The infusion of TBOA was conducted 0.5 (Anti+TBOA0.5(a)) or 2.5 h (Anti+TBOA2.5(a)) before spatial training (acquisition), immediately following training (consolidation; Anti+TBOA(b)), and 30 min prior to probe memory test (retrieval; Anti+TBOA(c)), respectively. (A) Schematic description of the experimental timeline. (B) Escape latency in the training phase and (C) the swim proximity score during the probe trial. Note the sample swimming traces demonstrating the swimming trajectories of the control, Anti+TBOA(b), and TBOA(b) groups rather than the Anti group superimposed on target quadrant. The triangle indicated the start point during probe trial. (* p

Article Snippet: A total of 418 male offspring from an average of 84 litters were randomly assigned to one of six groups: ( ) anti-proBDNF (second week), ( ) anti-proBDNF (fourth week), and ( ) anti-proBDNF (eighth week) groups received bilateral infusion of rabbit polyclonal anti-proBDNF antibody ( ; ) in the CA1 region of the HPC throughout the entire second postnatal week (PD2w, from PD8 to PD14), fourth postnatal week (PD4w, from PD22 to PD28), and eighth postnatal week (PD8w, from PD50 to PD56), respectively; ( ) control group was treated with the same volume of the vehicle (artificial cerebrospinal fluid, ACSF) throughout the whole PD2w (Con@2w), PD4w (Con@4w), and PD8w (Con@8w); ( ) Anti+TBOA group, which received infusion of anti-proBDNF antibody during the postnatal weeks, was bilaterally infused with DL-threo-β-benzyloxyaspartate (DL-TBOA) 0.5 or 2.5 h before spatial training [Anti+TBOA0.5(a) or Anti+TBOA2.5(a)], immediately following behavioral training [Anti+TBOA(b)] or 0.5 h before probe test [Anti+TBOA(c)]; and ( ) control group, which received infusion of ACSF during the postnatal weeks, was bilaterally infused with DL-TBOA 0.5 h before spatial training [TBOA(a)], immediately following behavioral training [TBOA(b)] or 0.5 h before probe test [TBOA(c)]; ( ) naive group was reared as the control group without the treatment.

Techniques: Activation Assay, Blocking Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Requirements of Postnatal proBDNF in the Hippocampus for Spatial Memory Consolidation and Neural Function

doi: 10.3389/fcell.2021.678182

Figure Lengend Snippet: GluN2B-dependent neural function is enhanced by TBOA. The Anti group was bilaterally infused with anti-proBDNF antibody into the CA1 region throughout the whole PD4w, whereas the Con group received the same volume of ACSF. Eight-week-old rats were selected for detecting hippocampal synaptic function in the Schaffer collateral-CA1 pathway immediately following TBOA (Anti+TBOA and TBOA groups), Ro25 (Ro25 group), or ACSF (Con and Anti groups) injection. (A) Input–output curves of fEPSP slopes. (B) PPF, a form of short-term plasticity, was measured and expressed as the ratio of fEPSPs2 to fEPSPs1. (* p

Article Snippet: A total of 418 male offspring from an average of 84 litters were randomly assigned to one of six groups: ( ) anti-proBDNF (second week), ( ) anti-proBDNF (fourth week), and ( ) anti-proBDNF (eighth week) groups received bilateral infusion of rabbit polyclonal anti-proBDNF antibody ( ; ) in the CA1 region of the HPC throughout the entire second postnatal week (PD2w, from PD8 to PD14), fourth postnatal week (PD4w, from PD22 to PD28), and eighth postnatal week (PD8w, from PD50 to PD56), respectively; ( ) control group was treated with the same volume of the vehicle (artificial cerebrospinal fluid, ACSF) throughout the whole PD2w (Con@2w), PD4w (Con@4w), and PD8w (Con@8w); ( ) Anti+TBOA group, which received infusion of anti-proBDNF antibody during the postnatal weeks, was bilaterally infused with DL-threo-β-benzyloxyaspartate (DL-TBOA) 0.5 or 2.5 h before spatial training [Anti+TBOA0.5(a) or Anti+TBOA2.5(a)], immediately following behavioral training [Anti+TBOA(b)] or 0.5 h before probe test [Anti+TBOA(c)]; and ( ) control group, which received infusion of ACSF during the postnatal weeks, was bilaterally infused with DL-TBOA 0.5 h before spatial training [TBOA(a)], immediately following behavioral training [TBOA(b)] or 0.5 h before probe test [TBOA(c)]; ( ) naive group was reared as the control group without the treatment.

Techniques: Injection

Journal: The Journal of Cell Biology

Article Title: BDNF and its pro-peptide are stored in presynaptic dense core vesicles in brain neurons

doi: 10.1083/jcb.201201038

Figure Lengend Snippet: α-BDNF, α-Myc, and α–pro-BDNF antibodies all generate similar staining patterns. (A) A schematic representation of the BDNF precursor pro-BDNF and the two cleavage products pro-peptide and BDNF. (B) Low-power view of a WT hippocampal section stained with anti-BDNF antibodies. Note the intense staining in the hilus of the DG and in SL of CA3, each of which contains the axon terminals of mossy fibers. (C and D) Higher magnification view of the DG (C) and CA3 region (D) of the cbdnf ko hippocampus stained with anti-BDNF. Note the absence of immunoreactive signals in all cellular and neuropil layers. GCL, granule cell layer; H, hilus; IML, inner molecular layer; PCL, pyramidal cell layer. (E) Bdnf-Myc hippocampi stained with Myc antibodies show a similar staining pattern to that produced by BDNF antibodies. (F and G) Note the absence of staining in the corresponding DG (F) and CA3 regions (G) of WT sections treated with Myc antibodies. (H) Polyclonal pro-BDNF antibodies yield a similar pattern to that of anti-BDNF. (I and J) The same antibodies do not produce an immunoreactive signal in hippocampal sections from cbdnf ko animals. (B, E, and H) Arrows denote the end bulb of the mossy fiber projection, which delineates CA3 and CA1. Note the relative lack of staining in CA1 in WT and Bdnf-Myc sections. Bars: (B, E, and H) 500 µm; (C, F, and I) 100 µm; (D, G, and J) 50 µm.

Article Snippet: Pro-BDNF was detected with a rabbit polyclonal antibody (anti–pro-BDNF; #ANT-006, batch AN-03; Alomone Labs) raised against the prodomain of BDNF protein (see ).

Techniques: Staining, Produced

Journal: Frontiers in Pharmacology

Article Title: Altered Trek-1 Function in Sortilin Deficient Mice Results in Decreased Depressive-Like Behavior

doi: 10.3389/fphar.2018.00863

Figure Lengend Snippet: Effect of deletion of Sort1 −/− gene on BDNF system. Western blot analyses and their corresponding histogram quantification of the expression of proteins involved in the BDNF system in brain extracts from WT and Sort1 −/− mice. (A) BDNF, (B) proBDNF, and (C) phospho-TrkB. ∗∗ p

Article Snippet: Rabbit polyclonal antibodies anti-TREK-1 and anti-proBDNF were from Alomone Labs (Israel).

Techniques: Western Blot, Expressing, Mouse Assay

Journal: PLoS ONE

Article Title: Fine-Tuning Roles of Endogenous Brain-Derived Neurotrophic Factor, TrkB and Sortilin in Colorectal Cancer Cell Survival

doi: 10.1371/journal.pone.0025097

Figure Lengend Snippet: Expression of BDNF and its receptors in human CRC cell lines. (A): RT-PCR of BDNF, its high (TrkB) and low (p75 NTR ) affinity receptors, TrkA, and TrkC from cells cultured in basal (10% FCS medium), WiDr (lane: 1), SW480 (lane: 2), SW620 (lane: 3), COLO 205 (lane: 4). GAPDH mRNA levels was used as an internal control. Positive control (lane 5) was the neuroblastoma cell line (IMR32) for BDNF, TrkB and p75 NTR ; and the erythromyeloblastoid leukemia cells (K562) for TrkA and C. A representative result of at least three to five independent experiments. (B) Comparison of TrkB95 and p75 NTR on total RNA extracted from WiDr cells cultured in 10% FCS or after 24 to 72 h serum deprivation (0% FCS). TrkB95 and p75 NTR mRNA/GAPDH mRNA quantification of band intensities evaluated by densitometry are shown above lanes and expressed in arbitrary units (mean of three independent experiments). (C) Same experiment: RT-PCR of TrkA and TrkC on total RNA extracted from WiDr cells cultured under basal culture conditions (10% FCS) and after 24–72 h of serum starvation in comparison to positive control (K562) (D) Expression of pro-BDNF and BDNF, full length TrkB 145 and truncated TrkB 95 and p75 NTR proteins in CRC cell lines cultured in 10% FCS. TrkA and TrkC were not detected. Actin was used as loading protein control. WiDr (lane: 1), SW480 (lane: 2), SW620 (lane: 3), COLO 205 (lane: 4). Positive control (lane: 5) were IMR32 cells for BDNF, pro-BDNF, TrkB and p75 NTR and K562 cells or TrkA and TrkC. A representative result of at least three independent experiments.

Article Snippet: The following Abs were used: rabbit anti-BDNF Ab (1 µg/ml; Santa Cruz Biotechnology), rabbit anti-pro-BDNF Ab (8 µg/ml; Alomone Labs), mouse anti-TrkB Ab (2.5 µg/ml; R & D Systems), rabbit anti-p75NTR (2 µg/ml; Santa Cruz Biotechnology) and goat anti-sortilin Ab (1 µg/ml; Santa Cruz Biotechnology).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Cell Culture, Positive Control

Journal: PLoS ONE

Article Title: Fine-Tuning Roles of Endogenous Brain-Derived Neurotrophic Factor, TrkB and Sortilin in Colorectal Cancer Cell Survival

doi: 10.1371/journal.pone.0025097

Figure Lengend Snippet: Membranous and cytoplasmic expression of BDNF and TrkB depending on culture conditions. Confocal microscopy of WiDr (A, B) and COLO 205 (C, D) cells, stained with an anti-BDNF Ab (red), anti-TrkB mAb (green) or both (overlay) cultured with 10% FCS (A, C) or after 24-h serum deprivation (B, D). Under basal culture conditions (10% FCS), TrkB and BDNF were sequestered in the cytoplasm (arrows) in WiDr (A) and COLO 205 (C) cells. The same staining patterns were obtained with the two other cell lines (data not shown). After serum starvation relocation to the cell membrane and colocalization of TrkB and BDNF (yellow in merged, arrows) were detected in WiDr (B) and COLO 205 (D). Images were representative for at least three to five independent experiments.

Article Snippet: The following Abs were used: rabbit anti-BDNF Ab (1 µg/ml; Santa Cruz Biotechnology), rabbit anti-pro-BDNF Ab (8 µg/ml; Alomone Labs), mouse anti-TrkB Ab (2.5 µg/ml; R & D Systems), rabbit anti-p75NTR (2 µg/ml; Santa Cruz Biotechnology) and goat anti-sortilin Ab (1 µg/ml; Santa Cruz Biotechnology).

Techniques: Expressing, Confocal Microscopy, Staining, Cell Culture

Journal: PLoS ONE

Article Title: Fine-Tuning Roles of Endogenous Brain-Derived Neurotrophic Factor, TrkB and Sortilin in Colorectal Cancer Cell Survival

doi: 10.1371/journal.pone.0025097

Figure Lengend Snippet: BDNF-TrkB promotes the cell survival of CRC cell lines. (A) Role of endogenous BDNF and its receptor TrkB on CRC cell proliferation: effects of exogenous BDNF and suppressing endogenous TrkB receptor on cell proliferation. The four cell lines were cultured for 24 h in FCS-free medium (FCS 10%, −) in the presence of exogenous BDNF (+), K252a (+) alone or in combination. Cell proliferation was determined by flow cytometry analysis using EdU Alexa Fluor 488. The data are presented as histograms of proliferating cells in relative units ± SEM of five independent experiments. *, p

Article Snippet: The following Abs were used: rabbit anti-BDNF Ab (1 µg/ml; Santa Cruz Biotechnology), rabbit anti-pro-BDNF Ab (8 µg/ml; Alomone Labs), mouse anti-TrkB Ab (2.5 µg/ml; R & D Systems), rabbit anti-p75NTR (2 µg/ml; Santa Cruz Biotechnology) and goat anti-sortilin Ab (1 µg/ml; Santa Cruz Biotechnology).

Techniques: Cell Culture, Flow Cytometry, Cytometry