Walter Peter, Alteheld Nils, Huth Julia, Roessler Gernot, Vobig Michael A
Department of Ophthalmology, RWTH Aachen University, Pauwelsstr. 30, 52074, Aachen, Germany.
Graefes Arch Clin Exp Ophthalmol. 2007 Apr;245(4):547-54. doi: 10.1007/s00417-006-0393-4. Epub 2006 Aug 11.
Our aim was to identify different states of energy metabolism in the perfused isolated vertebrate retina by simultaneous recordings of light-evoked changes of transretinal potential (TRP) and spectroscopic changes related to oxidation of pyridine nucleotides.
Isolated retinas were obtained from R.esc. and superfused. For each setting, three experiments were performed. Eighteen retinas were used. TRP as a response to light stimulation was recorded simultaneously with stimulus-induced transmission shift at 350 nm. These responses were recorded under normal conditions, under withdrawal of oxygen and glucose, and after addition of electron transport inhibitors and uncouplers of oxidative phosphorylation.
Under normal conditions, TRP was strongly correlated with transmission shift after light stimulation. Without oxygen, amplitudes of stimulus-related TRP decreased and response related increase of transmission at 350 nm was reduced. The retina showed a much higher general absorption of the analyzing light. During glucose lack, an overall increase in transmission at 350 nm could be observed. The addition of Amytal yielded a specific reduction of the OFF components. Cyanide attenuated the ON and OFF components. Dinitrophenol yielded a considerable loss of the electrical ON response. Transmission change was affected less than electrical signal. Chlorophenylhydrazine induced a reproducible reduction of electrical responses, with a relative increase of transmission shift at 350 nm.
In the isolated perfused retina, different states of energy metabolism and their specific impact on neuroretinal responses can be examined by simultaneous recordings of stimulus-related neuronal activity and spectroscopic changes.
我们的目的是通过同时记录光诱发的跨视网膜电位(TRP)变化和与吡啶核苷酸氧化相关的光谱变化,来识别灌注分离的脊椎动物视网膜中的不同能量代谢状态。
从R.esc.获取分离的视网膜并进行灌注。对于每种设置,进行了三个实验。使用了18个视网膜。在正常条件下、缺氧和缺葡萄糖条件下以及添加电子传递抑制剂和氧化磷酸化解偶联剂后,同时记录作为对光刺激反应的TRP和350nm处刺激诱导的透射率变化。
在正常条件下,TRP与光刺激后的透射率变化密切相关。无氧时,与刺激相关的TRP幅度降低,350nm处与反应相关的透射率增加减少。视网膜对分析光的总体吸收更高。在缺乏葡萄糖时,可观察到350nm处的透射率总体增加。添加阿米妥导致OFF成分特异性降低。氰化物减弱了ON和OFF成分。二硝基苯酚导致电ON反应显著丧失。透射率变化受影响小于电信号。氯苯肼诱导电反应可重复降低,350nm处的透射率变化相对增加。
在分离灌注的视网膜中,通过同时记录与刺激相关的神经元活动和光谱变化,可以研究不同的能量代谢状态及其对神经视网膜反应的特定影响。