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In-vivo-Imaging retinaler Zellapoptose nach akuter Lichtexposition

In vivo imaging of retinal cell apoptosis following acute light exposure

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Zusammenfassung

Hintergrund

Bisher war der Nachweis von retinaler Apoptose in der äußeren Körnerschicht nach akuter Lichtexposition auf histologische Untersuchungen [12] beschränkt. In dieser Studie untersuchen wir, ob programmierter Zelltod mittels der DARC-Technik (Detection-of-Apoptosing-Retinal-Cells-Technik) in vivo dargestellt werden kann.

Methoden

Die Augen von schwarzen Agouti-Ratten wurden mit blauem Licht (λ=405 nm; 3,2 mW/cm2) über 2 h bestrahlt. In-vivo-Imaging mittels konfokaler Scanning-Laser-Ophthalmoskopie wurde vor und direkt nach Lichtexposition sowie 24 h nach Dunkeladaptation durchgeführt. Anschließend wurde die Entwicklung von retinaler Zellapoptose unter Einsatz der DARC-Technik nach intravitrealer Gabe von fluoreszenzmarkiertem Annexin 5 untersucht.

Ergebnisse

Direkt nach Lichtexposition waren keine pathologischen Veränderungen durch In-vivo-Imaging zu erkennen. Hingegen wurde eine Netzhautverdünnung und die Entwicklung von retinaler Zellapoptose nach Dunkeladaptation einen Tag später im Bereich des vorher bestrahlten Areals nachgewiesen. Mittels konfokalem Live-Scanning durch die bestrahlte Netzhaut wurden die hyperfluoreszenten apoptotischen Zellen in der äußeren Netzhaut lokalisiert. Histologische Untersuchungen bestätigten die Entwicklung von Photorezeptorapoptose und Zellschäden im Bereich der äußeren Netzhaut.

Diskussion

Die DARC-Technik erlaubt die In-vivo-Darstellung von Photorezeptorzellapoptose in der äußeren Netzhaut. Hiermit eröffnen sich neue, vielsprechende Möglichkeiten zur Untersuchung des programmierten Photorezeptorzelltods, welcher bisher nur post mortem nachgewiesen werden konnte.

Abstract

Purpose

Outer nuclear apoptosis following acute light exposure has previously only been shown histologically. This study investigated whether in vivo detection with DARC (detection of apoptosing retinal cells) technology could identify cells undergoing apoptosis.

Methods

Acute blue light damage (λ=405 nm; 3.2 mW/cm2) was applied to eyes of dark Agouti rats over 2 h. In vivo retinal imaging using confocal scanning laser ophthalmoscopy was performed before and directly after light exposure as well as after 24 h of dark adaptation. Development of retinal cell apoptosis was then assessed using intravitreal fluorescent-labeled annexin-5 with DARC technology in vivo.

Results

Directly after light exposure, no pathological retinal changes were observed by in vivo imaging. However, retinal flattening and the development of apoptosis within the irradiated retina occurred 1 day later and following dark adaptation. Confocal live scanning through the exposed retina revealed hyperfluorescent apoptotic cells at the level of the outer retina. Histological analysis confirmed the occurrence of photoreceptor cell death and the development of cellular damage at the outer retina.

Discussion

This study confirms acute light-induced outer nuclear apoptosis using in vivo DARC technology. This may open new and promising ways to assess programmed cell death of the photoreceptor cells, which – until now – was possible only with postmortem analysis.

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Danksagung

Die Autoren danken Dr. Anthony Vugler, Dr. Carlos Gias und Vy Luong für wissenschaftliche und technische Unterstützung.

Interessenkonflikt

Der korrespondierende Autor gibt an, dass kein Interessenkonflikt besteht.

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Authors and Affiliations

  1. Universitäts-Augenklinik Bonn, Ernst-Abbe-Str. 2, 53127, Bonn, Deutschland

    S. Schmitz-Valckenberg

  2. UCL Institute of Ophthalmology, London, UK

    S. Schmitz-Valckenberg, L. Guo, W. Cheung, S.E. Moss & F.W. Fitzke

  3. UCL Institute of Ophthalmology, Western Eye Hospital, London, UK

    M.F. Cordeiro

Authors
  1. S. Schmitz-Valckenberg
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  2. L. Guo
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  3. W. Cheung
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  4. S.E. Moss
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  5. F.W. Fitzke
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  6. M.F. Cordeiro
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Correspondence to S. Schmitz-Valckenberg.

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Schmitz-Valckenberg, S., Guo, L., Cheung, W. et al. In-vivo-Imaging retinaler Zellapoptose nach akuter Lichtexposition. Ophthalmologe 107, 22–29 (2010). https://doi.org/10.1007/s00347-009-1952-y

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