1. Electroretinographic Assessment of Inner Retinal Signaling in the Isolated and Superfused Murine Retina.
- Author
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Albanna W, Lueke JN, Sjapic V, Kotliar K, Hescheler J, Clusmann H, Sjapic S, Alpdogan S, Schneider T, Schubert GA, and Neumaier F
- Subjects
- Animals, Barium Compounds pharmacology, Chlorides pharmacology, Dark Adaptation drug effects, Dark Adaptation radiation effects, Electroretinography drug effects, Electroretinography radiation effects, Mice, Models, Animal, Perfusion, Retinal Photoreceptor Cell Inner Segment drug effects, Retinal Photoreceptor Cell Inner Segment radiation effects, Signal Transduction drug effects, Signal Transduction physiology, Signal Transduction radiation effects, Dark Adaptation physiology, Electroretinography methods, Photic Stimulation methods, Retinal Photoreceptor Cell Inner Segment physiology
- Abstract
Purpose: Longer-lasting electroretinographic recordings of the isolated murine retina were initially achieved by modification of a phosphate-buffered nutrient solution originally developed for the bovine retina. During experiments with a more sensitive mouse retina, apparent model-specific limitations were addressed and improvements were analyzed for their contribution to an optimized full electroretinogram (ERG)., Material and Methods: Retinas were isolated from dark-adapted mice, transferred to a recording chamber and superfused with different solutions. Scotopic and photopic ERGs were recorded with white flashes every 3 minutes. The phosphate buffer (Sickel-medium) originally used was replaced by a carbonate-based system (Ames-medium), the pH of which was adjusted to 7.7-7.8. Moreover, addition of 0.1 mM BaCl
2 was investigated to reduce b-wave contamination by the slow PIII component typically present in the murine ERG., Results: B-wave amplitudes were increased by the pH-shift (pH 7.4 to pH 7.7) from 22.9 ± 1.9 µV to 37.5 ± 2.5 µV. Improved b-wave responses were also achieved by adding small amounts of Ba2+ (100 µM), which selectively suppressed slow PIII components, thereby unmasking more of the true b-wave amplitude (100.0% with vs. 22.2 ± 10.7% without Ba2+ ). Ames medium lacking amino acids and vitamins was unable to maintain retinal signaling, as evident in a reversible decrease of the b-wave to 31.8 ± 3.9% of its amplitude in complete Ames medium., Conclusions: Our findings provide optimized conditions for ex vivo ERGs from the murine retina and suggest that careful application of Ba2+ supports reliable isolation of b-wave responses in mice. Under our recording conditions, murine retinas show reproducible ERGs for up to six hours.- Published
- 2017
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