Longer lasting electroretinographic recordings from the isolated and superfused murine retina
Longer lasting electroretinographic recordings from the isolated and superfused murine retina
复制标题
DOI：
10.1007/s00417-009-1119-1
复制
发表时间：
2009-10-01
影响因子：
2.7
通讯作者：
Schneider, Toni
中科院分区：
文献类型：
--
作者：
Albanna, Walid;Banat, Mohammed;Schneider, Toni
对突变小鼠视网膜信号的分析已成为研究视网膜功能和疾病的有力工具。以前对分离的小鼠视网膜进行记录的尝试仅限于较短的时间段(约90分钟)。希望能够实现与在大鼠、兔子、猫和牛等大型脊椎动物中进行的记录相当的更持久的记录(长达10小时)。我们在多种不同的条件下对分离的小鼠视网膜进行了一系列记录，以确定该物种的最佳参数。我们使用了超融合脊椎动物视网膜实验，为此必须使用特定工具分离小鼠视网膜。为了提高视网膜光感受器和视网膜网络反应的敏感度和稳定性，使用了来自大鼠(生理钙离子)和牛(降低钙离子但增加磷酸盐缓冲能力)两种不同的营养液。此外，温度降低到27.5A℃，光强度增加十倍(63mlux)，以及增加的流速(2ml/min)提供了b波响应稳定超过3小时的条件。已知的钙通道拮抗剂(伊拉地平和NiCl2)被测试其拮抗经视网膜信号的效力。总之，分离的小鼠视网膜可以用作药理学测试系统，这为更好地理解视网膜信号提供了选择性基因失活的额外优势。
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 27.5A degrees 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.