Fig 1: Implanted enzymes penetrate the optic nerve and modify GAG chains.Zymosan was delivered to the retina via intravitreal injection and ARSB, ChABC, and control buffer were delivered to the lesioned optic nerve via implanted gelfoam scaffolds. (a) Schematic diagram showing delivery of ARSB, ChABC, or buffer to lesioned optic nerves via implanted gelfoam scaffold. Micrographs showing thresholded images of lesioned optic nerves treated with ChABC or buffer and stained with BE-123. Arrows indicate lesion site. Scale bar = 200 µm. (b) Graph showing quantification of BE-123 fluorescence intensity measured from insets centered at the lesion site. Statistical significance was determined by Student’s t-test. *p<0.05. (c) Schematic diagram showing delivery of ChABC to non-lesioned optics nerve via implanted gelfoam scaffold. Optic nerves were dissected at 1, 2, and 4 dpi and divided into four segments (#1–4). Western blot analysis shows enzyme-treated optic nerve segments stained with anti-BE-123 ‘stub’ antibody.
Fig 2: ARSB does not alter CS-56 or 2H6 immunoreactivity.(a) Micrographs showing insets from lesioned mouse optic nerve tissue analyzed by immunohistochemistry with CS-56 or 2H6. Mice received implants of carrying either ARSB or a control buffer at 3 dpc, and tissue was collected at 7 dpc. Scale bar = 50 µm. (b) Graph showing quantification of CS-56 and 2H6 fluorescence intensity measured from insets centered at the lesion site. Statistical significance was determined by Student’s t-test.
Fig 3: ARSB strongly enhances axon regeneration proximal to the lesion site.(a–b) Graphs showing the number of regenerating axons at distances distal to the lesion site, displayed as mean ± SEM. Data were collected from separate cohorts of mice where regeneration was quantified at 7, 14, and 28 days following ONC. (c) Graph showing the average increase in regenerating axons for each measured distance and time point, calculated by subtracting the number of axons in the buffer-treated group from the number of axons in the ARSB-treated group.
Fig 4: ARSB reverses neurite outgrowth inhibition caused by 4-sulfated CSPGs.(a) Micrographs showing CGNs treated with astrocyte conditioned medium containing TGF-β and either ARSB, ChABC, or no enzyme. Scale bar = 25 µm. (b) Plot showing length of longest neurite measured from β-III-tubulin stained neurons, displayed as median. Statistical significance was determined by two-way ANOVA with Tukey post-hoc test for multiple comparisons.
Fig 5: ARSB provokes muted immune response but does not alter astrocyte reactivity, glial scar size, or association of regenerating axons with astrocyte processes.(a) Micrographs showing Iba1 immunostaining at the optic nerve crush site for samples treated with Zymosan/CPT-cAMP and ChABC, Zymosan/CPT-cAMP and ARSB, Zymosan/CPT-cAMP and a control buffer, no treatment, and non-lesioned controls. Scale bar = 50 µm. (b) Graph showing quantification of Iba1 fluorescence intensity measured as % area of thresholded insets centered at the lesion site. Statistical significance was determined by Student’s t-test. *p<0.05, ***p<0.001. (c) Micrographs showing GFAP and GAP-43 immunostaining at the optic nerve crush site for samples treated with Zymosan/CPT-cAMP and either ARSB, ChABC, or a control buffer and analyzed at 7, 14, and 28 dpc. Arrows indicate lesion site. Scale bar = 100 µm. (d) Graph showing quantification of glial scar size measured as the area delineated by GFAP+ astrocytes at the optic nerve crush site. Statistical significance was determined by Student’s t-test. (e) Graph showing quantification of GFAP immunoreactivity at the optic nerve crush site. Statistical significance was determined by Student’s t-test.
Supplier Page from R&D Systems, a Bio-Techne Brand for Arylsulfatase B/ARSB Protein
Available conjugates: Sizes Available: 10 ug