Journal: Cell reports

Article Title: Leptin and IL-6 Family Cytokines Synergize to Stimulate Müller Glia Reprogramming and Retina Regeneration

doi: 10.1016/j.celrep.2014.08.047

Figure Lengend Snippet: (A) GFP and BrdU immunofluorescence shows Leptin synergizes with IL-11 and CNTF to stimulate GFP expression and MG proliferation in the uninjured retina 1016tuba1a:gfp fish, while Leptin, IL-11 or CNTF alone had little effect. Arrows point to MG-derived progenitors in top 3 panels. (B) Quantification of the effects of cytokines on MG proliferation when delivered individually or in combination to the uninjured retina; ***P<0.001 (combination vs individual), n=4 per group. (C, D) MO-mediated Ascl1a knockdown inhibits the synergistic effects of Leptin/IL-11 or Leptin/CNTF on GFP induction and MG proliferation; *P<0.05, n=3. (E, F) knockdown of Gp130 or Lepr inhibits the synergistic effects of Leptin/IL-11 on proliferation; ***P<0.001, n=4. Error bars, s.d. Scale bars, 50 µm (A, C, E). ONL, outer nuclear layer; INL, inner nuclear layer; GCL, ganglion cell layer.

Article Snippet: Recombinant human Leptin (gift of Amylin Pharmaceuticals Inc.), recombinant rat CNTF, recombinant human IL-6 and recombinant mouse IL-11 (R&D Systems) were reconstituted in PBS with 0.1% BSA, and 0.5–2 μl was injected intravitreally at the indicated concentration (See Table S2 for dissociation constants and estimates of intravitreal concentrations).

Techniques: Immunofluorescence, Expressing, Derivative Assay, Knockdown

Journal: Cell reports

Article Title: Leptin and IL-6 Family Cytokines Synergize to Stimulate Müller Glia Reprogramming and Retina Regeneration

doi: 10.1016/j.celrep.2014.08.047

Figure Lengend Snippet: (A) Intravitreal injection of Leptin/CNTF, or Leptin/IL-11 into the eye of gfap:stat3-gfp fish stimulates Stat3-GFP expression and BrdU incorporation throughout the uninjured retina’s INL, while Leptin, CNTF or IL-11 alone had little effect. Note that GFP reports activated p-Stat3 expression (Figure 1; Figure S2). White dots indicate autofluorescence unique to the green channel (see Figure S7K). The arrows point to MG-derived progenitors in the top 4 panels. (B) Quantification of Stat3-GFP+ cells in (A); ***P<0.001 (combination vs individual), n=4. (C) Quantification of BrdU+ cells in (A); ***P<0.001 (combination vs individual), n=4. (D) Western blot shows that retinal injury or intravitreal injection of cytokines into uninjured eye increases p-Stat3 expression. GS serves as loading control. (E) A model showing Leptin and IL-6 family cytokines synergize to stimulate MG reprogramming and retina regeneration via a Jak/Stat3 signaling pathway, which is essential for activating reprogramming genes like hbegf and ascl1a. Error bars, s.d. Scale bars, 50 µm. ONL, outer nuclear layer; INL, inner nuclear layer; GCL, ganglion cell layer.

Article Snippet: Recombinant human Leptin (gift of Amylin Pharmaceuticals Inc.), recombinant rat CNTF, recombinant human IL-6 and recombinant mouse IL-11 (R&D Systems) were reconstituted in PBS with 0.1% BSA, and 0.5–2 μl was injected intravitreally at the indicated concentration (See Table S2 for dissociation constants and estimates of intravitreal concentrations).

Techniques: Injection, Expressing, BrdU Incorporation Assay, Derivative Assay, Western Blot, Control