Journal: Acta Neuropathologica Communications

Article Title: Brain-derived neurotrophic factor supports pericyte and vascular homeostasis in the aging brain

doi: 10.1186/s40478-025-02181-y

Figure Lengend Snippet: Aging reduces vasculature and pericyte expression of PDGFRβ in the brain. Brain tissue from C57BL/6J mice at 6, 12 and 24 months of age was stained for collagen type IV ( A) and quantified for total length and branch points of microvessels ( B – E ; t -test, n = 7 and 4–5 for the 6–12 months and 24 months groups, respectively). Cerebral microvessels were isolated from 12 to 24-month-old mice and homogenized for Western blot analysis of pericyte markers PDGFRβ and CD13, and protein levels of phosphorylated and total Akt and Erk1/2 ( F – L ; t -test, n = 8—9 and 7—8 per group for the 6–12 months and 24 months groups, respectively). In following experiments, transcriptomic analysis was performed using a single-cell sequencing dataset. Volcano plots show up- and down-regulated DEGs in the brains of 18-month-old mice compared with 2-month-old mice ( M ; NS, genes with non-significant changes). The bar chart displays the top five significantly enriched KEGG pathways ranked by − log 10 ( p -value) (bar length). The size of adjacent circles corresponds to the number of DEGs in each pathway. KEGG pathway analysis indicated that the down-regulated genes are associated with PI3K-AKT signaling pathway ( N )

Article Snippet: After blocking with 50 μg/ml CD16/CD32 antibody (clone 2.4G2; BioXCell, Lebanon, USA), brain cells were stained with PE-conjugated rat monoclonal antibody against mouse PDGFRβ (clone APB5; Miltenyi Biotec GmbH).

Techniques: Expressing, Staining, Isolation, Western Blot, Sequencing

Journal: Acta Neuropathologica Communications

Article Title: Brain-derived neurotrophic factor supports pericyte and vascular homeostasis in the aging brain

doi: 10.1186/s40478-025-02181-y

Figure Lengend Snippet: Deficiency of neuronal BDNF reduces vasculature and pericytes in the brain. Ten-month-old C57BL/6 mice with (ko) and without (wt) knockout of Bdnf gene in neurons for 3 months were stained for collagen type IV and quantified for the vasculature ( A ). Deficiency of neuronal BDNF significantly reduced the total length of cerebral vessels and tended to decrease the density of branch points of vessels ( B and C ; t -test, n = 6–7 per group). The brain sections were also co-stained for PDGFRβ and CD31. PDGFRβ-positive pericytes were counted and adjusted by the length of CD31-positive vessels ( D ). Deficiency of neuronal BDNF significantly reduced the number of pericytes ( E ; t -test, n = 4 per group). Additionally, microvessels were isolated from brains and detected with Western blot for pericyte markers and relevant signaling molecules ( F and I ). Deficiency of neuronal BDNF decreased protein levels of PDGFRβ but not CD13 ( G and H ; t -test, n = 6 per group). BDNF deficiency also reduced the phosphorylation of Akt, but not Erk1/2 ( J and K ; t -test, n = 4–9 per group)

Article Snippet: After blocking with 50 μg/ml CD16/CD32 antibody (clone 2.4G2; BioXCell, Lebanon, USA), brain cells were stained with PE-conjugated rat monoclonal antibody against mouse PDGFRβ (clone APB5; Miltenyi Biotec GmbH).

Techniques: Knock-Out, Staining, Isolation, Western Blot, Phospho-proteomics

Journal: Acta Neuropathologica Communications

Article Title: Brain-derived neurotrophic factor supports pericyte and vascular homeostasis in the aging brain

doi: 10.1186/s40478-025-02181-y

Figure Lengend Snippet: Deficiency of astrocyte BDNF reduces vasculature and pericytes in the brain. Brain homogenates from 10 month-old C57BL/6 mice with (ko) and without (wt) knockout of Bdnf gene in astrocytes for 3 months were detected for protein levels of BDNF ( A ). Knockout of Bdnf gene significantly reduced mature BDNF but not pro-BDNF ( B and C ; t -test, n = 9–11 per group). Brain sections were then stained for collagen type IV and quantified for the vasculature ( D ). Deficiency of astrocyte BDNF significantly reduced both the length and density of branch points of cerebral vessels (E and F; t -test, n = 8 per group). Additionally, microvessels were isolated from brains and detected with Western blot for pericyte markers and relevant signaling molecules ( G and J ). Deficiency of astrocyte BDNF decreased the protein level of PDGFRβ, but not CD13 ( H and I ; t -test, n = 11–12 per group), and reduced the phosphorylation of both Akt and Erk1/2 ( K and L ; t -test, n = 4–9 per group)

Article Snippet: After blocking with 50 μg/ml CD16/CD32 antibody (clone 2.4G2; BioXCell, Lebanon, USA), brain cells were stained with PE-conjugated rat monoclonal antibody against mouse PDGFRβ (clone APB5; Miltenyi Biotec GmbH).

Techniques: Knock-Out, Staining, Isolation, Western Blot, Phospho-proteomics

Journal: Acta Neuropathologica Communications

Article Title: Brain-derived neurotrophic factor supports pericyte and vascular homeostasis in the aging brain

doi: 10.1186/s40478-025-02181-y

Figure Lengend Snippet: BDNF acts directly on cultured pericytes. Human pericyte and endothelial cell lines were cultured. The transcription levels of BDNF receptor genes, NTRK2 and NGFR , were measured with real-time PCR. Both cells expressed NGFR gene at a significantly higher level than NTRK2 gene ( A and B ; t -test, n = 4 per group. Four experiments were independently repeated). Western blot showed expression of TrkB in the pericyte cell line ( C ). Cultured pericytes were then treated with BDNF at 0, 10, 50, and 100 ng/ml for 24 h. Western blot was used to detect the protein level of PDGFRβ, showing that BDNF treatments significantly increase PDGFRβ expression ( D and E ; One-way ANOVA followed by Bonferroni post hoc test, n = 10 per group for 0, 10 and 50 concentrations and n = 5 for 100 ng/ml concentration. Ten experiments were independently repeated), as well as to determine the phosphorylation levels of both Akt and Erk1/2 ( D , F and G ; One-way ANOVA followed by Tukey post hoc test, n = 4 or 3 per group. Four and three experiments were independently repeated for Akt and Erk1/2, respectively). Pericytes were also treated with BDNF at different concentrations in the presence of 1 µM Akt inhibitor VIII. The protein level of PDGFRβ as detected by Western blot was not altered by the treatments of BDNF ( H and I ; One-way ANOVA, p > 0.05, n = 4 per group. Four experiments were independently repeated). In additional experiments, endothelial cells were treated with BDNF at 0, 50 and 100 ng/ml for 24 h. BDNF significantly up-regulates transcription of PDGF-B , but not CD62p and CD31 genes ( J – L ; One-way ANOVA followed by Tukey post hoc test, n = 4 per group. Four experiments were independently repeated)

Article Snippet: After blocking with 50 μg/ml CD16/CD32 antibody (clone 2.4G2; BioXCell, Lebanon, USA), brain cells were stained with PE-conjugated rat monoclonal antibody against mouse PDGFRβ (clone APB5; Miltenyi Biotec GmbH).

Techniques: Cell Culture, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Concentration Assay, Phospho-proteomics

Journal: International journal of molecular sciences

Article Title: Thyroid Hormone Transporters MCT8 and OATP1C1 Are Expressed in Projection Neurons and Interneurons of Basal Ganglia and Motor Thalamus in the Adult Human and Macaque Brains.

doi: 10.3390/ijms24119643

Figure Lengend Snippet: Figure 4. Expression of MCT8 and OATP1C1 in blood vessels and brain barriers in the human and macaque basal ganglia and adjacent choroid plexus. (A) Representative brightfield photomicro- graph shows immunostaining for MCT8 in the human putamen. Note that an MCT8 immunoposi- tive signal is observed along the capillary wall (red arrowhead), fibers (green arrowhead), and “bump-on-a-log” morphology pericytelike cells (white arrowhead). (B–H) Representative confocal microscope compositions from multiple-stained sections for MCT8 (green), the endothelial marker UEA-I (red), and the vascular and pericyte biomarker PDGFR-β (purple) in human and macaque caudate nucleus. Merged image (E,H) shows the colocalization of all signals. (B–E) Coexpression of MCT8, UEA-I, and PDGFR-β is observed in a vessel, while coexpression of MCT8 and PDGFR- β but not UEA-I is observed in a capillary-associated pericyte (white arrowheads) in humans. (F–H) Coexpression of MCT8 and PDGFR-β in a vessel and pericytelike cells (white arrowheads) in macaques. Counterstaining with DAPI (blue) shows nuclei of all cells. (I,J) Representative bright- field photomicrographs show immunostaining for MCT8 (I) and OATP1C1 (J) in the macaque choroid plexus at the lateral ventricle. Black arrowheads point to ependymocytes. Cd: caudate nu- cleus, Put: putamen, PDGFR-β: platelet-derived growth factor receptor-β, UEA-I: Ulex europaeus agglutinin-I. Scale bar = 10 µm (A–H) and 50 µm (I,J).

Article Snippet: The antibody against PDGFR-β (IHC 1:200/IF 1:100, AF385, R&D systems, Minneapolis, MN, USA) was used to label all vascular mural cells [92]; the lectin UEA-I was used to label blood vessel endothelial cells [93]; anti-DRD1 (IHC 20 μg/mL/IF 25 μg/mL, MAB8276, R&D systems) labeled D1-MSN in the direct pathway and anti-DRD2 (IHC 20 μg/mL/IF 25 μg/mL, MAB9266, R&D systems) labeled D2-MSN in the indirect pathway [27]; the antibody against the calcium-binding protein CALB (IHC 1:2000/IF 1:1000, #C9848, Sigma-Aldrich) was used to label striatal GABAergic interneurons [29] and matrix projection cells in the thalamus [37,38,94]; the antibody against the calcium-binding protein CALR (IHC 1:2000/IF 1:1000, 6B3, Swant, Surry Hills, Australia) labeled GABAergic interneurons in neostriatum [29]; the antibody against ChAT (IHC 1:500/IF 1:250, AB144P, Merck Millipore, Burlington, MA, USA) labeled cholinergic interneurons in neostriatum [29] and cholinergic neurons in nucleus basalis of Meynert [54]; the antibody against PARV (IHC 1:2000/IF 1:1000, #P3088, SigmaAldrich) labeled striatal GABAergic interneurons [29], GABAergic neurons in GPe, GPi and SNr [32,33,95], projection neurons in STN, and core projection cells in thalamus [37]; the antibody against nNOS (IHC 15 μg/mL/IF 10 μg/mL, AF2416, R&D systems) labeled GABAergic interneurons in neostriatum [31]; the antibody against SOM (IHC 1:500/IF 1:300, sc-55565, Santa Cruz Biotechnology) labeled GABAergic interneurons in neostriatum [30]; and finally the antibody TYH (IHC 1:400/IF 1:200, sc-25269, Santa Cruz Biotechnology) labeled dopaminergic neurons in SNc [95].

Techniques: Expressing, Immunostaining, Microscopy, Staining, Marker, Biomarker Discovery, Derivative Assay

Journal: Cells

Article Title: Extracellular Vesicles from Skeletal Muscle Cells Efficiently Promote Myogenesis in Induced Pluripotent Stem Cells

doi: 10.3390/cells9061527

Figure Lengend Snippet: Pericyte characterization. ( A ) Representative histograms indicating the percentage of alkaline phosphatase (ALP + ), platelet derived growth factor receptor-beta (PDGFRβ + ), MCAM, melanoma cell adhesion molecule (CD146 + ), (αSMA + ), Neuron/glial antigen 2 (NG2 + ), HCAM, homing cell adhesion molecule (CD44 + ) and NCAM, neural-cell adhesion molecule (CD56 + ) positivity (black peaks) determined by flow cytometry in pre-sorted cells isolated from muscular biopsy (n = 4). Matched isotypes were used as negative controls (grey peaks). ( B ) Representative gating strategy for ALP + and CD56 − cell sorting (n = 3). Cells were first gated for cell size (side light scatter SSC-A vs. forward light scatter FSC-H) and vitality (Live Qdot-525-A). The muscular cell gate was further analyzed for singlets (SSC-A vs. SSC-H) and their expression for ALP and CD56. Pericytes, ALP + and CD56 − were then sorted from this gated population. The lower set of four plots confirmed the efficiency of the sorting. ( C ) Representative post-sorting histograms for key pericyte markers after two passages in vitro, indicating an enhanced expression of ALP, NG2, PDGFRβ, CD146, and CD44. ( D ) Sorted pericytes stained for ALP showing fibroblast colony-forming units (CFU-F) when seeded at low confluence. Scale bar represents 300 μm. ( E ) Immunofluorescence labeling for NG2 (red) and the co-staining for PDGFRβ (green) and αSMA (magenta) on sorted ALP + CD56 − cells. Nuclei were stained with DAPI. Scale bar represents 50 μm. ( F ) Representative fluorescence image for myosin heavy chain (MHC) (red), validating the differentiation of sorted pericytes toward skeletal muscle phenotype. Scale bar represents 100 μm. ( G ) Illustrative images of human umbilical vein endothelial cells (HUVEC) in co-culture with pericytes displaying the formation of capillary-like networks with HUVEC labeled for von Willebrand factor (vWF; magenta), and GFP + pericytes. Nuclei were identified by DAPI (blue). Scale bar represents 100 μm. Tubular structures were photographed at 5× magnification and quantified by the angiogenesis analyzer ImageJ tool. Total segment length, total mesh area and total branching length exhibited significant differences between HUVEC alone and in co-culture with pericytes, as shown in the graphs.

Article Snippet: Muscle pericytes were labelled with the following conjugated antibodies: anti-alkaline phosphatase-Cy5 (BD Pharmingen), anti-CD45-FITC/CD14-PE (BD Biosciences, San Jose, CA, USA), anti-NG2-PE (BD Pharmingen), anti-CD56-APC (NCAM; BD Biosciences), anti-CD146-Cy5 (MCAM; R&D Systems, Minneapolis, MN, USA), anti-PDGF-R-beta-FITC (R&D Systems), and anti-CD44-APC (BD Pharmingen).

Techniques: Derivative Assay, Flow Cytometry, Isolation, FACS, Expressing, In Vitro, Staining, Immunofluorescence, Labeling, Fluorescence, Co-Culture Assay

Journal: Journal of Cerebral Blood Flow & Metabolism

Article Title: Differing associations between Aβ accumulation, hypoperfusion, blood–brain barrier dysfunction and loss of PDGFRB pericyte marker in the precuneus and parietal white matter in Alzheimer's disease

doi: 10.1177/0271678X17690761

Figure Lengend Snippet: Platelet-derived growth factor receptor-β (PDGFRB) loss and blood–brain barrier (BBB) breakdown in relation to disease severity (i.e. Braak tangle stage), APOE genotype, and cerebral amyloid angiopathy (CAA) in the precuneus in AD. Bar charts showing (a) reduced PDGFRB in Braak tangle stage V–VI (end-stage) compared to Braak tangle stage 0–II. ( P < 0.01) (b) reduced PDGFRB level in APOE ε 3.4 ( P < 0.01) and APOE ε 4.4 ( P < 0.05) compared with APOE ε 2.3 individuals and (c) reduced PDGFRB level in severe CAA compared to absent CAA ( P < 0.01). Bar charts showing (d) increased fibrinogen level in Braak tangle stage V–VI (end stage) compared to Braak stage 0–II ( P < 0.01), (e) no significant difference in fibrinogen level in relation to APOE genotype and (f) increased fibrinogen level in moderate CAA compared to absent CAA ( P < 0.01). The bars indicate the mean and SEM. CAA severity scores adapted from Olichney et al.: , 0 = absent, 1 = mild, 2 = moderate, 3 = severe.

Article Snippet: PDGFRB level was measured by sandwich ELISA (duoset, Cat no DYC385, R&D systems, Oxford, UK).

Techniques: Derivative Assay

Journal: Journal of Cerebral Blood Flow & Metabolism

Article Title: Differing associations between Aβ accumulation, hypoperfusion, blood–brain barrier dysfunction and loss of PDGFRB pericyte marker in the precuneus and parietal white matter in Alzheimer's disease

doi: 10.1177/0271678X17690761

Figure Lengend Snippet: Platelet-derived growth factor receptor-β (PDGFRB) loss and blood–brain barrier breakdown are associated with hypoperfusion of the precuneus. (a) Scatterplot showing a strong positive correlation between PDGFRB and MAG:PLP1 (r = 0.24, P = 0.006), i.e. the lowest PDGFRB levels were in samples with least preservation of MAG relative to PLP1. (b) Scatterplot showing a negative correlation between PDGFRB level and VEGF level in the precuneus (r = −0.26, P = 0.029), i.e. the lowest PDGFRB levels were in samples with greatest elevation in VEGF. (c) Scatterplot showing a negative correlation between fibrinogen level and MAG:PLP1 (r = −0.30, P = 0.022) and (d) Scatterplot showing a strongly positive correlation between fibrinogen and VEGF level (r = 0.49, P < 0.0001). Each point in the scatterplots indicates a single AD (red circle) or control (blue square) brain. The best-fit linear regression lines and 95% confidence intervals are superimposed.

Article Snippet: PDGFRB level was measured by sandwich ELISA (duoset, Cat no DYC385, R&D systems, Oxford, UK).

Techniques: Derivative Assay, Preserving, Control

Journal: Journal of Cerebral Blood Flow & Metabolism

Article Title: Differing associations between Aβ accumulation, hypoperfusion, blood–brain barrier dysfunction and loss of PDGFRB pericyte marker in the precuneus and parietal white matter in Alzheimer's disease

doi: 10.1177/0271678X17690761

Figure Lengend Snippet: Reduced oxygenation in the parietal white matter in AD associated with blood–brain barrier (BBB) breakdown despite concomitant increase in pericytes. (a) Scatterplot showing strong negative correlation between platelet-derived growth factor receptor-β (PDGFRB) and myelin-associated glycoprotein:proteolipid protein-1 (MAG:PLP1) ratio in white matter (WM) (r = −0.31, P = 0.004). (b) Scatterplot showing positive correlation between platelet-derived growth factor receptor-β (PDGFRB) level and vascular endothelial growth factor (VEGF) level (r = 0.19, P = 0.019). (c) Scatterplot showing very strong negative correlation between fibrinogen and MAG:PLP1 (r = −0.48, P < 0.0001). (d) Scatterplot showing very strong positive correlation between fibrinogen and VEGF level in the white matter (r = 0.42, P < 0.0001). Each point in the scatterplots indicates a single AD (red circle) or control (blue square) brain. The best-fit linear regression lines and 95% confidence intervals are superimposed.

Article Snippet: PDGFRB level was measured by sandwich ELISA (duoset, Cat no DYC385, R&D systems, Oxford, UK).

Techniques: Derivative Assay, Control

Journal: Frontiers in Immunology

Article Title: Unveiling spatial complexity in solid tumor immune microenvironments through multiplexed imaging

doi: 10.3389/fimmu.2024.1383932

Figure Lengend Snippet: Immunophenotyping panel for multiplexed tissue imaging of cancer.

Article Snippet: PDGFR β , MAB1263 , 200 , FAB1263T-100UG , FITC , RnD Systems.

Techniques: Imaging

Journal: Human Mutation

Article Title: Temperature as a Key Modulator: Investigating Phosphorylation Patterns of p.Asn666 PDGFRB Variants and Their Role in Downstream Signaling

doi: 10.1155/humu/6664372

Figure Lengend Snippet: (a) All PDGFR β substitutions (p.Asn666Lys, p.Asn666Ser, p.Asn666His, and p.Asn666Tyr) increase basal phosphorylation at 37°C. (b) Exposure to 32°C for 6 h further increases phosphorylation in the p.Asn666Tyr, p.Asn666Lys, and p.Asn666His substitutions. Total phosphorylated PDGFR β levels were measured by ELISA. Nontransduced immortalized fibroblasts were referred to as “nt,” while fibroblasts transduced with PDGFRB wild-type vector were denoted as “WT PDGFRB .” (a) One-way ANOVA with Dunnett's test compared wild-type PDGFRB and variants at 37°C (⁣ ∗∗∗∗ p < 0.0001). (b) Cells were incubated at 32°C or 37°C, and two-way ANOVA with Šídák's test compared phosphorylation between temperatures for each variant (⁣ ∗ p < 0.05; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001).

Article Snippet: In brief, ELISA analysis was conducted using a DuoSet IC PDGFR β kit (# DYC1767-2, R&D Systems, Minneapolis, MN) following the guidelines provided by the manufacturer.

Techniques: Phospho-proteomics, Enzyme-linked Immunosorbent Assay, Transduction, Plasmid Preparation, Incubation, Variant Assay

Journal: Human Mutation

Article Title: Temperature as a Key Modulator: Investigating Phosphorylation Patterns of p.Asn666 PDGFRB Variants and Their Role in Downstream Signaling

doi: 10.1155/humu/6664372

Figure Lengend Snippet: Effect of reduced temperature (32°C) and ligand stimulation on PDGFR β -specific tyrosine residue phosphorylation. Representative immunoblots showing phosphorylation of PDGFR β at p-Y-740, p-Y-751, p-Y-771, p-Y-1009, and p-Y-1021. At 37°C, p.Asn666Lys and p.Asn666Ser variants exhibited nearly identical signal intensity, whereas p.Asn666His and p.Asn666Tyr showed a distinct pattern. At 32°C, all variants exhibited a variant-specific increase in phosphorylation. PDGF stimulation (10 ng/mL) altered phosphorylation patterns, increasing the upper PDGFR β band, likely representing the fully mature, membrane-associated receptor. Full immunoblots, also overexposed, are shown in Figures and .

Article Snippet: In brief, ELISA analysis was conducted using a DuoSet IC PDGFR β kit (# DYC1767-2, R&D Systems, Minneapolis, MN) following the guidelines provided by the manufacturer.

Techniques: Residue, Phospho-proteomics, Western Blot, Variant Assay, Membrane

Journal: Human Mutation

Article Title: Temperature as a Key Modulator: Investigating Phosphorylation Patterns of p.Asn666 PDGFRB Variants and Their Role in Downstream Signaling

doi: 10.1155/humu/6664372

Figure Lengend Snippet: Effects of lower physiological temperature (32°C) on PDGFR β downstream signaling. (a) Immunoblots showing temperature-dependent changes in p-AKT-Ser473, p-AKT-Thr308, p-STAT1, p-STAT3-Tyr705, p-STAT6, and p-PLC γ 1, in immortalized fibroblasts expressing the four p.Asn666 variants. GAPDH served as a loading control. Full immunoblots are shown in Figure . (b,c) Quantification of downstream signaling is presented as the ratio of phosphorylated protein to GAPDH. (b) Normalized values for each variant compared to the wild type at 37°C. (c) Rations for each variant at 32°C versus 37°C. Symbols represent fold changes: “-” (≥ 30% decrease), “≈” (−30% to +50%), “(+)” (150–200%), “+” (2- to 4-fold), “++” (4- to 10-fold), “+++” (10- to 50-fold), “++++” (> 50-fold). Bar charts summarizing these data are provided in Figures and .

Article Snippet: In brief, ELISA analysis was conducted using a DuoSet IC PDGFR β kit (# DYC1767-2, R&D Systems, Minneapolis, MN) following the guidelines provided by the manufacturer.

Techniques: Western Blot, Expressing, Control, Variant Assay