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Y. I. Leiderman, T. Hisatomi, D. P. Biss, C. Alt, C. P. Lin, J. W. Miller; Imaging Apoptotic Cell Death in the Murine Retina in vivo Following Experimental Retinal Detachment. Invest. Ophthalmol. Vis. Sci. 2009;50(13):3173.
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© ARVO (1962-2015); The Authors (2016-present)
Retinal detachment (RD) in humans and experimental animal models induces photoreceptor cell death. Apoptosis has been shown to be the predominant mechanism of retinal cell death following RD, as assessed by ultrastructural and biochemical analyses of post-mortem tissue. Histologic analyses of dynamic processes are constrained by virtue of depicting intercellular interactions at a single time-point. We set out to detect and observe apoptotic cell death in the retina in vivo following retinal detachment in a murine model.
Experimental RD was induced in C57BL/6 mice via injection of sodium hyaluronate into the subretinal space. Apoptotic cells in the neuro-retina were labeled via injection of Annexin V conjugated to fluorescein isothiocyanate (FITC) into the subretinal space on day 0 and 3 following RD. In vivo retinal imaging was performed via scanning laser ophthalmoscopy (SLO) with an adaptive optics (AO) system specifically designed for imaging the murine eye. The AO system included a microelectromechanical system membrane mirror and Shack-Hartmann wavefront sensor. Retinal angiography was performed with Evans Blue using a 635 nm laser for excitation. A 491 nm laser was used for excitation of FITC-labeled cells in the retina.
SLO angiography with AO allowed fine resolution of the retinal vasculature and delineated the presence of retinal detachment. Punctate foci of Annexin V-FITC hyperfluorescence were observed within detached retina at 3 days following the induction of RD, but not immediately following induction of RD with either sodium hyaluronate or Annexin V-FITC on day 0.
SLO with AO allows simultaneous imaging of fluorescently labeled cells and resolution of RD morphology and vascular patterns. To our knowledge, this is the first observation of apoptotic retinal cell death in vivo following experimental RD.
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