Ryba N, Uhl R
Max-Planck-Institut für biophysikalische Chemie, Göttingen, Federal Republic of Germany.
Exp Brain Res. 1989;74(3):618-24. doi: 10.1007/BF00247364.
A method is described which allows the in vitro dark adaptation of rod photoreceptors from cattle eyes, enucleated under ambient light in the slaughterhouse. Without in vitro dark adaptation these eyes are light adapted and cannot be used for certain delicate biochemical studies and for an electrophysiological characterisation of rod responses. The method is very simple and yields large amounts of dark adapted retinal material, allowing experiments that require bulk amounts of photoreceptor cells. The only source of dark adapted photoreceptors so far have been retinae from dark adapted laboratory animals, which had to be killed and processed under infrared light. Eye cups were opened under red light as soon as possible after their enucleation. Their vitreous humor was removed and their retina thoroughly rinsed with ringer's. Then the eye cup was placed in a moist, light-tight box, where dark adaptation took place. Photoreceptors could thus be kept alive for more than 24 h without showing signs of deterioration. Humidity and free access of oxygen to the retina were the only prerequisites for their survival. The physiological intactness of the photoreceptors and their degree of dark adaptation was demonstrated by measuring mass receptor potentials (ERGs). A simple device is described which can be used for the electrophysiological characterisation of these eyes.
本文描述了一种方法,该方法能够使从屠宰场在环境光下摘除的牛眼视杆光感受器进行体外暗适应。如果不进行体外暗适应,这些眼睛处于明适应状态,无法用于某些精细的生化研究以及视杆反应的电生理特性研究。该方法非常简单,能产生大量暗适应的视网膜材料,从而可以开展需要大量光感受器细胞的实验。迄今为止,暗适应光感受器的唯一来源一直是来自暗适应实验动物的视网膜,这些动物必须在红外光下处死并进行处理。眼球在摘除后尽快在红光下打开。去除其玻璃体并用林格氏液彻底冲洗视网膜。然后将眼球放入一个潮湿、不透光的盒子中进行暗适应。这样光感受器可以存活超过24小时而不出现退化迹象。湿度和视网膜对氧气的自由接触是其存活的唯一前提条件。通过测量群体感受器电位(视网膜电图)来证明光感受器的生理完整性及其暗适应程度。本文还描述了一种可用于这些眼睛电生理特性研究的简单装置。