Longer lasting electroretinographic recordings from the isolated and superfused murine retina
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Analysis of retinal signaling in mutant mice has become a powerful tool for studying retinal function and disease. Previous attempts to record from isolated mouse retina have been limited to short time periods (about 90 min). It would be desirable to achieve longer-lasting recordings comparable to those that have been performed in larger vertebrates such as rat, rabbit, cat, and bovine (up to 10 h). We performed a series of recordings from isolated mouse retina under a number of different conditions in order to determine the optimal parameters for this species.
We used a superfused vertebrate retina assay, for which the murine retina had to be isolated with specific tools. Subsequently, the ERG recordings were optimized for nutrient solution, incubation temperature, and flash light intensity.
To improve the sensitivity and stability of photoreceptor and retinal network responses from the isolated and superfused murine retina, two different nutrient solutions from rat (physiological Ca2+) and bovine (reduced Ca2+ but increased phosphate buffering capacity) were used. Further, a temperature reduced to 27.5°C, a light intensity ten-fold increased (63 mlux), and an increased flow rate (2 ml/min) provided conditions under which the b-wave response was stable for more than 3 hours. Well-known Ca2+ channel antagonists (isradipine and NiCl2) were tested for their potency to antagonize transretinal signalling.
In conclusion, the isolated murine retina can be used as a pharmacological testing system, which provides the additional advantage of selective gene inactivation for better understanding of retinal signalling.
KeywordsMurine ERG Isolated vertebrate retina a-wave b-wave Light sensitivity Dihydropyridine NiCl2 Oxygen
We acknowledge the support from Prof. Dr. J. Hescheler, chairman of the Institute for Neurophysiology, Cologne. The work was financially supported by the Köln Fortune Program/Faculty of Medicine, University of Köln by a Promotionsstipendium to W.A. and M.B., and by the Center of Molecular Medicine Cologne/Zentrum für Molekulare Medizin Köln (Bundesministerium für Bildung, Wissenschaft, Forschung und Technologie, Förderkennzeichen 01 KS 9502, to TS, MW and JH).
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