FIGURE

Fig. 4

ID
ZDB-FIG-150129-12
Publication
Zhao et al., 2014 - Leptin and IL-6 Family Cytokines Synergize to Stimulate Müller Glia Reprogramming and Retina Regeneration
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Fig. 4

IL-6 Family Cytokines Signaling through Gp130 Are Necessary and Sufficient for Retina Regeneration

(A) In situ hybridization and immunofluorescence show that gp130, il-11a, crlf1a, and clcf1 are expressed in BrdU+ MG-derived progenitors localized to the injury site.

(B) qPCR quantifies il-6 family gene induction in MG-derived progenitors (FACS purified from 1016tuba1a:gfp fish retinas at 4 dpi) relative to MG from uninjured retina (FACS purified from uninjured gfap:gfp fish retinas). p < 0.05, n = 3.

(C and D) Gp130 knockdown inhibits the generation of BrdU+ MG-derived progenitors at 4 dpi. Control (Ctl) or gp130-targeting MOs were electroporated into the retina at the time of injury and the fish received an i.p. injection of BrdU 3 hr before sacrifice on 4 dpi. p < 0.001, n = 4.

(E) qPCR showing Gp130 knockdown inhibits injury-dependent induction of reprogramming genes at 2 dpi. p < 0.05, p < 0.01, n = 4.

(F) Intravitreal injection of recombinant mammalian IL-6-like cytokines into the uninjured eye of 1016tuba1a:gfp fish stimulates GFP expression and BrdU incorporation in MG throughout the retina’s INL. Intravitreal injection of PBS/BSA did not stimulate GFP expression or BrdU incorporation. The green fluorescence above the ONL in the top left-hand panel is autofluorescence unique to the green channel (see Figure S7K).

(G) Quantification of BrdU+ cells following intravitreal injection of recombinant mammalian IL-6-like cytokines (n = 3).

Error bars, SD. In (A) and (C), the asterisks mark the injury site (needle poke). In (A), (C), and (F), arrows point to MG-derived progenitors. Scale bars, 20 µm (A and C) and 50 µm (F). GCL, ganglion cell layer. Primers are listed in Table S1. See also Figures S5 and S6.

Expression Data
Genes:
Fish:
Condition:
Anatomical Term:
Stage: Adult

Expression Detail
Antibody Labeling
Phenotype Data
Fish:
Condition:
Knockdown Reagent:
Observed In:
Stage: Adult

Phenotype Detail
Acknowledgments
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