Journal: British journal of pharmacology

Article Title: Neurotrophic interactions between neurons and astrocytes following AAV1-Rheb(S16H) transduction in the hippocampus in vivo.

doi: 10.1111/bph.14882

Figure Lengend Snippet: FIGURE 4 Elevation of CNTF production in hippocampal astrocytes via activation of the BDNF/TrkB/mTORC1 signalling pathway. (a) Immunohistochemical staining showing increased CNTF expression in glia‐like cells of the hippocampal CA1 region of CON, AAV1‐GFP‐ injected, and AAV1‐Rheb(S16H)‐injected rats (black arrows). Scale bars, 500 μm (inset 20 μm). (b) Double immunofluorescence staining against GFAP (green) and CNTF (red) showing the co‐localization of these two markers in the CA1 region of the hippocampus of CON and AAV1‐Rheb (S16H)‐injected rats. Scale bars, 100 μm (inset 10 μm). (c, d) Western blot analysis of the levels of CNTF expression in the hippocampus of AAV1‐ GFP‐injected or AAV1‐Rheb(S16H)‐injected rats, with or without BDNF neutralizing antibody (B.NA) or TrkB neutralizing antibody (T.NA). Differences among groups were evaluated by Kruskal–Wallis test (c) or one‐way ANOVA (d) and Tukey's post hoc analysis. *P < .05 versus CON and #P < .05 versus Rheb(S16H) alone (n = 5 for each group). (e) Immunohistochemical staining showing increased CNTF expression in glia‐like cells of the hippocampal CA1 region of recombinant BDNF‐injected rats (black arrows). Double immunofluorescence staining against GFAP (green) and CNTF (red) showing the co‐localization of these two markers in the CA1 region of the hippocampus of recombinant BDNF‐injected rats. Scale bar, 50 μm. (f) Western blot analysis of CNTF. Differences between groups were evaluated by Student's unpaired t test. *P < .05 versus CON (n = 5 for each group). (g–i) Measurement of CNTF concentration in the conditioned medium (CM) of recombinant BDNF‐treated astrocyte cultures using ELISA kits. (g) Schematic of the experimental design for measuring the levels of CNTF in hippocampal astrocyte cultures. (h) Histogram showing the dose‐dependent effects of recombinant BDNF on CNTF release in astrocyte cultures. Differences among groups were evaluated by Kruskal–Wallis test and Tukey's post hoc analysis. *P < .05 versus CON (n = 5 for each group). (i) Quantitative results showing the CNTF concentration after the treatment of astrocyte cultures with recombinant BDNF (80 ng·ml−1), with or without GNF‐5837 (10 μM) or rapamycin (80 nM). Differences among groups were evaluated by one‐way ANOVA and Tukey's post hoc analysis. *P < .05 versus CON and #P < .05 versus recombinant BDNF alone (n = 5 for each group)

Article Snippet: To quantify the CNTF released in the BDNF‐treated astrocyte culture medium, we used commercially available ELISA kits according to the manufacturer's protocol (Cat No. DY557; R&D Systems).

Techniques: Activation Assay, Immunohistochemical staining, Staining, Expressing, Injection, Double Immunofluorescence Staining, Western Blot, Recombinant, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Biomedical Materials

Article Title: Ciliary neurotrophic factor mediated growth of retinal ganglion cell axons on PGS/PCL scaffolds

doi: 10.1088/1748-605x/ad1bae

Figure Lengend Snippet: Figure 1. Experimental groups (Group I–IV) including, Group I with the REs were cultured on TCP, Group II with REs seeded on PGS/PCL, Group III with the REs cultured on CNTF immobilized PGS/PCL and, Group IV with the REs circumscribed around the 3D PVA enclosure on PGS/PCL in the presence of CNTF gradient.

Article Snippet: ELISA measurements were performed according to the manufacturer’s instruction (CNTF DuoSet ELISA; R&D Systems Inc., USA; DY557 15 plates).

Techniques: Cell Culture

Journal: Biomedical Materials

Article Title: Ciliary neurotrophic factor mediated growth of retinal ganglion cell axons on PGS/PCL scaffolds

doi: 10.1088/1748-605x/ad1bae

Figure Lengend Snippet: Figure 2. ATR-FTIR spectroscopy of ‘as fabricated’ PGS/PCL (blue), PLO-LA coated PGS/PCL (red) and, CNTF immobilised PLO-LA coated PGS/PCL (green).

Article Snippet: ELISA measurements were performed according to the manufacturer’s instruction (CNTF DuoSet ELISA; R&D Systems Inc., USA; DY557 15 plates).

Techniques: Spectroscopy

Journal: Biomedical Materials

Article Title: Ciliary neurotrophic factor mediated growth of retinal ganglion cell axons on PGS/PCL scaffolds

doi: 10.1088/1748-605x/ad1bae

Figure Lengend Snippet: Figure 3. The amount of CNTF released after 6 h (D0), 10 d (D10) and 20 d (D20) in the conditioned medium of CNTF immobilized PGS/PCL scaffolds quantified by ELISA measurements.

Article Snippet: ELISA measurements were performed according to the manufacturer’s instruction (CNTF DuoSet ELISA; R&D Systems Inc., USA; DY557 15 plates).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Biomedical Materials

Article Title: Ciliary neurotrophic factor mediated growth of retinal ganglion cell axons on PGS/PCL scaffolds

doi: 10.1088/1748-605x/ad1bae

Figure Lengend Snippet: Figure 5. The MAP2 staining and DAPI images of REs on day 10 and day 20 of culture on control (Group I) (top), unmodified PGS/PCL polymeric scaffold (Group II) (second from the top), on CNTF immobilized PGS/PCL (Group III) scaffold (second from the bottom) and, on PGS/PCL with CNTF gradient positioned within the 3D enclosure (Group IV) (bottom images).

Article Snippet: ELISA measurements were performed according to the manufacturer’s instruction (CNTF DuoSet ELISA; R&D Systems Inc., USA; DY557 15 plates).

Techniques: Staining, Control

Journal: Biomedical Materials

Article Title: Ciliary neurotrophic factor mediated growth of retinal ganglion cell axons on PGS/PCL scaffolds

doi: 10.1088/1748-605x/ad1bae

Figure Lengend Snippet: Figure 6. The average neurite lengths of neurons in µm on (a) day 10 and (b) day 20 on control (Group I), unmodified PGS/PCL polymeric scaffold (Group II), on CNTF immobilized PGS/PCL (Group III) scaffold and, on PGS/PCL with CNTF gradient positioned within the 3D enclosure (Group IV).

Article Snippet: ELISA measurements were performed according to the manufacturer’s instruction (CNTF DuoSet ELISA; R&D Systems Inc., USA; DY557 15 plates).

Techniques: Control

Journal: Biomedical Materials

Article Title: Ciliary neurotrophic factor mediated growth of retinal ganglion cell axons on PGS/PCL scaffolds

doi: 10.1088/1748-605x/ad1bae

Figure Lengend Snippet: Figure 8. The REs on the designed ex vivo model (a) MAP2 staining with the CNTF gradient decreasing from the left to the right of the image; the CNTF immobilized on PGS/PCL are placed inside the 3D printed enclosure, and (b) a binary processed image of the MAP2 stained image; (c)–(d) density distribution profile of the REs showing the left, image (c), and the right, image (d), of the explant.

Article Snippet: ELISA measurements were performed according to the manufacturer’s instruction (CNTF DuoSet ELISA; R&D Systems Inc., USA; DY557 15 plates).

Techniques: Ex Vivo, Staining