Ccr1, Ccr3, Ccr5, and Cxcr6 Mediate Chemoattraction of MSCs toward Media Conditioned by Degenerating Retinae
(A and B) Quantification of migrated MSCs toward the conditioned media from either (A) PBS- versus NMDA-injected eyecups or (B) WT versus rd10 P18 eyecups. Number of migrated cells is expressed as fold change relative to control samples (PBS-injected or WT). Data are presented as mean ± SD (n ≥ 3). A Mann-Whitney test was used for statistical analysis. (C) Quantification of migrated MSCs toward the conditioned media from human retinae cultured in control medium, NMDA-containing medium, or from RP-affected retinae. The number of migrated cells is expressed as fold change relative to control retinae. Data are presented as mean ± SD (n ≥ 3). (D and E) A Mann-Whitney test was used for statistical analysis. CC and CXC inflammatory chemokine profiles from either (D) PBS- versus NMDA-injected retinae (24 h post-infection [hpi]) or (E) WT versus rd10 P18 retinal lysates. Pixel intensity is directly proportional to the total amount of protein in the sample. Orange arrows point to representative chemokines whose abundance is increased in degenerating retinae (NMDA or rd10), relative to controls (PBS or WT). Data are presented as mean ± SD from n = 2 independent experiments. (F and G) qRT-PCR of CC and CXC inflammatory chemokines from (F) NMDA-damaged (24 hpi) and (G) rd10 (P18) retinae. Transcript levels are expressed as fold change relative to control (PBS-injected or WT) retinae. Data are presented as mean ± SD (n ≥ 3). A Mann-Whitney test was used for statistical analysis. (H) Scheme summarizing the interactions among chemokine receptors and ligands of interest. Ccr1, Ccr3, and Ccr5 are bound by Ccl5, Ccl6, and Ccl12. Cxcr2 is bound by Cxcl1, Cxcl2, and Cxcl5. Cxcr6 is bound by Cxcl16. (I and J) Quantification of migrated MSCs toward the conditioned media from either (I) NMDA-damaged (24 hpi) or (J) rd10 (P18) retinae, in the presence of the inhibitors of Ccrs (Ccr1, Ccr3, and Ccr5 [iCcrs]), Cxcr2 (iCxcr2), Cxcr6 (iCxcr6) or Ccrs + Cxcrs (Ccr1, Ccr3, Ccr5 [iCcrs]; Cxcr2 and Cxcr6 [iCxcrs]). Number of migrated cells is expressed as fold change to control (vehicle, DMSO). Data are presented as minimum (Min) to maximum (Max) boxes (with line at median) from n ≥ 3 independent experiments. A Mann-Whitney test was used for statistical analysis.