dl phenylalanine  (Chem Impex International)

Journal: Signal Transduction and Targeted Therapy

Article Title: Astrocytic monoamine oxidase B (MAOB)–gamma-aminobutyric acid (GABA) axis as a molecular brake on repair following spinal cord injury

doi: 10.1038/s41392-025-02398-2

Figure Lengend Snippet: Astrocytic MAOB and GABA impede functional and tissue recovery, whereas astrocytic BDNF is critical for survival after SCI. a Experimental timelines using MAOB KO, aKO, and aOE. b BMS score of each group (MAOB WT, WT + SCI, MAOB KO, and KO + SCI) for a total of 11 weeks (1 week before and 10 weeks after the surgical operation). c , d BMS score of each group for aKO ( c ), and aOE ( d ) for a total of 11 weeks. e , g , i EC staining of spinal cord tissues from MAOB KO ( e ), aKO ( g ), and aOE ( i ) at PI 10w. f , h , j Total cord area (left) and myelinated area (right) of MAOB KO ( f ), aKO ( h ), and aOE ( j ) mice in each group, as determined by EC staining. The myelinated area was normalized to that of the WT in group the MAOB KO and aOE. k Confocal images of the injured areas of MAOB KO (left), aKO (middle), and aOE (right) stained with anti-MAP2 (red), anti-GFAP (white), and anti-MAOB (green) antibodies, and DAPI (blue) at PI 10w. Each yellow box in the merged images indicates the magnified region of interest. l – n Mean intensity of the MAP2 (top) and GFAP-positive MAOB (bottom) in each group for MAOB KO ( l ), aKO ( m ), aOE ( n ) at PI 10w. o Experimental timeline using BDNF gKO and aKO. p BMS score of each group for a total of 9 weeks. q Survival curves of each group. r EC staining of cross (top) and longitudinal (bottom) sections of spinal cord tissues in each group (CTL 80 gKO+SCI, BDNF gKO+SCI, CTL aKO+SCI, and BDNF aKO+SCI) at PI 8w. s Total cord, myelinated, 81 and gray matter areas did not show any difference among the groups. t Schematic figure showing the calculation method for the Cobb angle. u Calculated the Cobb angle of each group at PI 8w (left) and representative image showing spine deformity of BDNF gKO+SCI (right). All data are expressed as the means ± S.E.M.s * P < 0.05; ** P < 0.01; *** P < 0.001; n.s., not significant. #, ##, ###, and δ indicate P < 0.05, P < 0.01, P < 0.001, and P < 0.0001, respectively, for the linear mixed model

Article Snippet: B6- Maob em1Cjl /Ibs ( Maob floxed) was generated from Cyagen Biosciences (Guangzhou, China).

Techniques: Functional Assay, Staining

Journal: Signal Transduction and Targeted Therapy

Article Title: Astrocytic monoamine oxidase B (MAOB)–gamma-aminobutyric acid (GABA) axis as a molecular brake on repair following spinal cord injury

doi: 10.1038/s41392-025-02398-2

Figure Lengend Snippet: MAOB inhibition with KDS2010 causes functional/tissue recovery and neuroregeneration. a Experimental timelines for 8-week-old rats subjected to the SCI operation and treatment with the MAOB inhibitor, KDS2010, from 2 weeks after (subacute) SCI. b BBB score of each group (Sham+V, SCI + V, SCI + KDS 2w) for a total of 11 weeks (1 week before and 10 weeks after the surgical operation). c Hindlimb movement in the open field test for SCI + V and SCI + KDS 2w. The group labels indicate the treatment start point. d Percentage of hindlimb steps without slipping in each group with subacute phase KDS2010 treatment. e Schematic of the ladder rung test apparatus. Created with BioRender.com. f BBB score of each group with three different doses of KDS2010 (Sham+V, SCI + V, SCI + KDS 10 mpk, 20 mpk, and 30 mpk) for a total of 11 weeks. All doses of KDS2010 were administered from the subacute phase. g EC staining of spinal cord tissues at each dose at PI 10w. h Total cord area and myelinated area of each group via EC staining. i Confocal images of the injured area in each group stained with anti-MAP2 (red), anti-GFAP (white), and anti-MAOB (green) antibodies at PI 10w. Each yellow box in the merged images indicates the magnified region of interest. j Mean intensity of the MAP2 (left) and GFAP-positive MAOB (right) in each group. k TEM images of spinal cord tissues at PI 2w and 10w in SCI + V and SCI + KDS 2w. l Calculated method for g-ratio. m Calculated g-ratio in each group. The blue shading indicates the optimal range of g-ratio (0.790 ± 0.005). n Schematic figure showing DNA constructs in each virus and the result of recombination. o Schematic figure showing the virus injection sites at 1 mm proximal and distal the injured area. p Experimental timeline using rats with the SCI operation, virus injection, and subacute phase treatment with KDS2010 at 10 mpk. q Confocal images of the injured area in SCI + V stained with anti-GFP (green), mCh (red) antibodies and DAPI (blue) at PI 10w. r Confocal images of the injured area (left) and 3D surface rendering images of mCh signals in SCI + KDS 2w at PI 10w. Shaded areas in ( b , d , and k ) indicate the duration of KDS2010 administration. All the data are expressed as the means ± S.E.M.s * P < 0.05; ** P < 0.01; *** P < 0.001; n.s., not significant. #, ##, and ### indicate P < 0.05, P < 0.01, P < 0.001, and P < 0.0001, respectively, for the linear mixed model

Article Snippet: B6- Maob em1Cjl /Ibs ( Maob floxed) was generated from Cyagen Biosciences (Guangzhou, China).

Techniques: Inhibition, Functional Assay, Staining, Construct, Virus, Injection

Journal: Signal Transduction and Targeted Therapy

Article Title: Astrocytic monoamine oxidase B (MAOB)–gamma-aminobutyric acid (GABA) axis as a molecular brake on repair following spinal cord injury

doi: 10.1038/s41392-025-02398-2

Figure Lengend Snippet: MAOB inhibition with KDS2010 reduces astrocytic GABA levels and enhances proBDNF and TrkB expression after SCI. a Differential interference contrast (DIC) images of the dorsal horn of the spinal cord (top and middle) and a magnified view of a whole-cell patch-clamped lamina II neuron (bottom). The yellow boxes indicate the magnified region of interest. b Representative traces of GABA A receptor-mediated tonic GABA current in each group (Sham+V, SCI + V, and SCI + KDS 2w). The dashed lines (gray) and double-headed arrows (purple and brown) indicate baseline shifts (I GABA and I Tonic ) with bath application of GABA (10 μM, green bar) and bicuculline (Bic, 50 μM, orange bar). c Tonic GABA current density (top, left), GABA-induced full activation current density (top, right), frequency (bottom, left) and amplitude (bottom, right) of spontaneous inhibitory postsynaptic currents (sIPSCs) in each group. d Confocal images of the injured areas stained with anti-proBDNF (green), anti-GABA (red), anti-NeuN (magenta), and anti-GFAP (white) antibodies at PI 10w in SCI + V and SCI + KDS 2w. e Mean intensity of GABA (left), NeuN-positive proBDNF (middle), GFAP-positive proBDNF (right). f Western blotting of BDNF and TrkB in Sham+V, SCI + V, and SCI + KDS 2w at PI 10w. g Quantification of BDNF (left) and TrkB (right) expression levels in Western blotting. β-actin was used as a control for protein amount. h Experimental timeline for quantitative real-time PCR with GABA (100 μM) or Bic (50 μM) treatment of primary cultured spinal cord astrocytes at 14 days in vitro (DIV). i Relative (comparative Ct) BDNF expression level of each drug treatment condition (GABA, GABA+Bic, and Bic). j Representative images of control and GABA-treated spinal cord astrocytes. k Western blotting of proBDNF in control and GABA-treated condition. l Quantification of proBDNF in Western blotting. β-actin was used as a control for protein amount. m Confocal images of the injured areas stained with anti-TrkB (green), DAPI (blue), and each Tau, MBP, or GFAP (red) antibodies at PI 10w. KDS2010 treatment was initiated at PI 2w and continued daily until PI 10w. Group labels indicate the treatment start point. n Mean intensity of TrkB in the Tau-positive neurons (left), MBP-positive oligodendrocytes (middle), and GFAP-positive astrocytes (right) at PI 10w. All the data are expressed as the means ± S.E.M.s * P < 0.05; ** P < 0.01; *** P < 0.001; n.s., not significant

Article Snippet: B6- Maob em1Cjl /Ibs ( Maob floxed) was generated from Cyagen Biosciences (Guangzhou, China).

Techniques: Inhibition, Expressing, Activation Assay, Staining, Western Blot, Control, Real-time Polymerase Chain Reaction, Cell Culture, In Vitro

Journal: Signal Transduction and Targeted Therapy

Article Title: Astrocytic monoamine oxidase B (MAOB)–gamma-aminobutyric acid (GABA) axis as a molecular brake on repair following spinal cord injury

doi: 10.1038/s41392-025-02398-2

Figure Lengend Snippet: MAOB inhibition with KDS2010 facilitates tissue recovery after SCI in nonhuman primates. a Location of the SCI and representative longitudinal tissue sections at PI 5w in cynomolgus macaques ( Macaca fascicularis ). The red arrow indicates hematoma, and the yellow arrows indicate the cavities. Created with BioRender.com. b Experimental timelines using 3–4-year-old cynomolgus macaques subjected to the SCI operation and treatment with KDS2010 from 1 day after (acute) SCI. c Confocal images of the injured areas stained with anti-MAP2 (red), anti-GFAP (white), and anti-MAOB (green) antibodies, and DAPI (blue) at PI 5w in each group (Sham+V, SCI + V, SCI + KDS 3 mpk, and SCI + KDS 10 mpk). d Mean intensity of the MAP2 (left) and GFAP-positive MAOB (right) in each group. e Methodology illustration for blood chemistry analysis with nonhuman primate SCI model. Created with BioRender.com. f Blood chemistry and liver function data before and after KDS2010 treatment, showing the levels of serum creatine kinase (IU/L), albumin (ALB), gamma-glutamyl transferase (GGT, IU/L), total serum bilirubin (TBIL, mg/dL), alkaline phosphatase (ALP, IU/L), alanine transaminase (ALT, g/dL), and aspartate transaminase (AST, IU/L) at 2 days, 2 weeks, and 5 weeks post-injury. All the data are expressed as the means ± S.E.M.s * P < 0.05; ** P < 0.01; *** P < 0.001; n.s., not significant

Article Snippet: B6- Maob em1Cjl /Ibs ( Maob floxed) was generated from Cyagen Biosciences (Guangzhou, China).

Techniques: Inhibition, Staining

Journal: Experimental neurobiology

Article Title: Generation of Astrocyte-Specific MAOB Conditional Knockout Mouse with Minimal Tonic GABA Inhibition.

doi: 10.5607/en22016

Figure Lengend Snippet: Fig. 1. Generation of Maob floxed and astrocyte-specific MAOB cKO mouse lines. (A) Schematic diagram of Maob location on mouse X chromosome (top) and construction of Maob floxed allele using the CRIPSR-Cas9 technique (bottom). (B) Construct of Maob floxed allele in Maob floxed mouse (B6-Maob em1Cjl/Ibs) with primer sets for genotyping (F1-R1, F2-R2) and sequencing (F1, R2) for each loxP site. (C) Genotyping result of homozygote, heterozygote, WT, and distilled water (DW) as no template control using F1-R1 primer (left) and F2-R2 primer (right). Red dotted boxes and lines in- dicate extracted DNA bands for sequencing in (D) and (E). (D) Sequencing result of loxP site in upstream of exon 2 using F1 primer. Orange and grey sequences indicate deleted and inserted intronic sequences from the original Maob, respectively. (E) Sequencing result of loxP site in downstream of exon 2 using R2 primer. Orange and grey sequences indicate deleted and inserted intronic sequences from the original Maob, respectively. (F) Schematic diagram showing generation of astrocyte-specific MAOB cKO mice by crossing Maob floxed mice (B6-Maob em1Cjl/Ibs, X’X, WT) with hGFAP-CreER T2 (B6-Tg(GFAP-cre/ERT2)13Kdmc, XY, TG) (left), and genotyping results of Maob floxed::hGFAP-CreER T2 (X’Y, TG) with F1-R1, F2-R2, and two pairs of primers for hGFAP-CreER T2 with no template control of each primer set (middle), and construct of Maob allele in astrocyte-specific MAOB cKO mouse (right).

Article Snippet: Generation of Maob floxed mouse line We requested the generation of Maob floxed mouse line to the Cyagen Biosciences (Guangzhou, China).

Techniques: Construct, Sequencing, Control

Journal: Experimental neurobiology

Article Title: Generation of Astrocyte-Specific MAOB Conditional Knockout Mouse with Minimal Tonic GABA Inhibition.

doi: 10.5607/en22016

Figure Lengend Snippet: Fig. 2. MAOB and GABA levels in the cerebellum are reduced in astrocyte-specific MAOB cKO mice. (A) Experimental scheme and timeline using Maob floxed::hGFAP-CreER T2 mice. (B) Representative confocal images of MAOB (green), GFAP (magenta) and DAPI (blue) fluorescence in sagittal slices of the cerebellum in sunflower oil-injected MAOB control and tamoxifen-injected MAOB cKO mice. ML, molecular layer; GCL, granule cell layer. (C, D) Quan- tification of MAOB intensity in GFAP-positive cells in the GCL (C; MAOB control, 2186 voxels; MAOB cKO, 695 voxels; Mann-Whitney test, p<0.0001) and in the ML (D; MAOB control, 3455 voxels; MAOB cKO, 9916 voxels; Mann-Whitney test, p<0.0001). (E, F) Quantification of MAOB intensity in GFAP-negative areas in the GCL (E; MAOB control, 27 pixels; MAOB cKO, 22 pixels; Mann-Whitney test, p=0.8034) and in the ML (F; MAOB control, 22 pixels; MAOB cKO, 19 pixels; Mann-Whitney test, p=0.1084). (G) Representative SIM images of MAOB (green) and GFAP (magenta) in sagittal slices of cerebellum in MAOB control (left) and astrocytic MAOB cKO mice (right). White boxes, magnified regions. Insets, magnified and rotated 3-dimensional (3D) images. (H) Representative images for GABA (green) and GFAP (magenta) in sagittal slices of cerebellum in MAOB control and astrocytic MAOB cKO mice. (I, J) Quantification of GABA intensity in GFAP-positive cells in the GCL (I; MAOB control, 73 voxels; MAOB cKO, 84 voxels; Mann-Whitney test, p<0.0001) and in the ML (J; MAOB control, 68 voxels; MAOB cKO, 63 voxels; Unpaired t test, p<0.05). (K, L) Quantification of GABA intensity in GFAP-negative areas in the GCL (K; MAOB control, 20 pixels; MAOB cKO, 22 pixels; Mann-Whitney test, p=0.1508) and in the ML (L; MAOB control, 26 pixels; MAOB cKO, 33 pixels; Mann-Whitney test, p=0.5119). In violin plots, the center line denotes the median value, while upper and lower lines denote the first quartile and third quartile, respectively. Data are presented as mean±SEM. ****p<0.0001; *p<0.05; n.s., not significant.

Article Snippet: Generation of Maob floxed mouse line We requested the generation of Maob floxed mouse line to the Cyagen Biosciences (Guangzhou, China).

Techniques: Fluorescence, Injection, Control, MANN-WHITNEY

Journal: Experimental neurobiology

Article Title: Generation of Astrocyte-Specific MAOB Conditional Knockout Mouse with Minimal Tonic GABA Inhibition.

doi: 10.5607/en22016

Figure Lengend Snippet: Fig. 4. MAOB and GABA levels in the striatum are reduced in astrocyte-specific MAOB cKO mice. (A) Experimental scheme and timeline using Maob floxed::hGFAP-CreER T2 mice. (B) Representative confocal images of MAOB (green), S100β (magenta) in coronal slices of striatum in sunflower oil- injected MAOB control and tamoxifen-injected MAOB cKO mice. (C) Quantification of MAOB intensity in S100β-positive cells in the striatum (MAOB control, 130316 voxels; MAOB cKO, 89368 voxels; Mann-Whitney test, p<0.0001). (D) Quantification of MAOB intensity in S100β-negative areas in the striatum (MAOB control, 75 pixels; MAOB cKO, 73 pixels; Mann-Whitney test, p=0.5655). (E) Representative SIM images of MAOB (green) and S100β (magenta) in coronal slices of striatum in MAOB control (left) and astrocytic MAOB cKO mice (right). White boxes, magnified regions. Insets, magni- fied and rotated 3D images. (F) Representative images for GABA (green), S100β (magenta) and DAPI (blue) in coronal slices of striatum in MAOB con- trol and astrocytic MAOB cKO mice. (G) Quantification of GABA intensity in S100β-positive cells in the striatum (MAOB control, 28 voxels; MAOB cKO, 30 voxels; Unpaired t test, p<0.01. (H) Quantification of GABA intensity in S100β-negative areas in the striatum (MAOB control, 85 pixels; MAOB cKO, 75 pixels; Mann-Whitney test, p=0.1110). In violin plots, the center line denotes the median value, while upper and lower lines denote the first quartile and third quartile, respectively. Data are presented as mean±SEM. ****p<0.0001; **p<0.01; n.s., not significant.

Article Snippet: Generation of Maob floxed mouse line We requested the generation of Maob floxed mouse line to the Cyagen Biosciences (Guangzhou, China).

Techniques: Injection, Control, MANN-WHITNEY

Journal: Experimental neurobiology

Article Title: Generation of Astrocyte-Specific MAOB Conditional Knockout Mouse with Minimal Tonic GABA Inhibition.

doi: 10.5607/en22016

Figure Lengend Snippet: Fig. 6. Schematic illustration of generation (left) and characterization (right) of astrocyte-specific MAOB cKO mice. Tamoxifen-injected astrocyte- specific MAOB cKO mice show minimal tonic GABA inhibition with reduced levels of MAOB expression and GABA content in astrocytes, compared to sunflower oil-injected MAOB control mice.

Article Snippet: Generation of Maob floxed mouse line We requested the generation of Maob floxed mouse line to the Cyagen Biosciences (Guangzhou, China).

Techniques: Injection, Inhibition, Expressing, Control