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Integrin survival signaling is down-regulated in RGC in the retina 1 day after ischemic injury. A.

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posted on 19.02.2013 by Andrea Rachelle C. Santos, Raul G. Corredor, Betty Albo Obeso, Ephraim F. Trakhtenberg, Ying Wang, Jamie Ponmattam, Galina Dvoriantchikova, Dmitry Ivanov, Valery I. Shestopalov, Jeffrey L. Goldberg, Mary Elizabeth Fini, Michaela Livia Bajenaru

(i ) Western blot of retinal extracts of ischemic, RIRI (+), and control (−) rat eyes 1 day post-injury (n = 3). A representative example of at least 5 independent experiments is presented. (ii) Densitometric analysis of western blotting with all samples normalized to β-actin. Error bars, SD. Student’s t test. *p<0.05. B. β1 integrin expression is decreased in RGC 1 day post-RIRI. Immunohistochemistry with β1 integrin antibodies (red) in frozen retinal sections from control (a, b, c), and ischemic retinas (d, e, f), in which RGCs were retrogradely labeled with FG (green; a, d) shows reduced β1 integrin expression in ischemic eyes in comparison with control eyes (red; b, e) specifically in RGC (yellow; merged; c, f). Nuclei are stained with DAPI (blue; c, f). (n = 12). GCL = ganglion cell layer, INL = inner nuclear layer. Scale bars: 10 µm. C. FAK activation is reduced in RGC 1 day after ischemia. Immunohistochemistry with FAK (purple), and P-FAK [pY397] (red) antibodies in frozen retinal sections of control (a, b, c), and ischemic retinas (d, e, f) at 1 day post-RIRI, in which RGCs were retrogradely labeled with FG (yellow; a, d) shows significantly decreased expression of the activated, phospho [pY397] FAK in ischemic eyes in comparison with control eyes (red; c, f), while FAK expression remains unchanged (purple; b, e). White arrows identify FG labeled RGCs in the control retina (CTRL) expressing high levels of both FAK, and P-FAK [pY397] (a, b, c). Blue arrows point toward RGCs in the ischemic retina (RIRI) that have markedly decreased expression of P-FAK [pY397], while FAK expression is the same (d, e, f). (n = 12). GCL = ganglion cell layer, INL = inner nuclear layer. Scale bars: 10 µm.

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