Huang J C, Voaden M J, Marshall J
Institute of Ophthalmology, University of London, UK.
Curr Eye Res. 1990 Feb;9(2):151-62. doi: 10.3109/02713689008995201.
Procedures for regenerating visual pigment and restoring phototransduction have been established with freshly isolated, bleached rat retinas. Phosphatidyl choline liposomes containing a 500 microM mixture of retinal isomers, including the 9-cis and 11-cis forms, were employed and the results compared with dark-adapted retinas, incubated similarly but without retinal. The following were recovered in a 60 min incubation, rhodopsin (plus isorhodopsin) to 91% of the original rhodopsin concentration, 87% of cGMP and 89% of PIII amplitude at saturation. PIII amplitude vs. log intensity curves gave values of n between 0.6 and 1/2.0 and sigma between 85 and 439 quanta/micron 2. Human retinas, ranging from 18 to 58 hours postmortem and treated as above, also produced photoresponses. Of the 7 retinas studied so far, rhodopsin has been regenerated to 0.1-0.35 nmol/mg protein, cGMP to 23.5-49.2 pmol/mg protein, and PIII to 20-50 microV: in some cases a b-wave was also seen. Values of n varied between 0.6 and 1.0, and sigma between 132 and 3700 quanta/micron 2. PIII responses were also seen after retinas, approximately 30 hours postmortem, were incubated for a further 24 hours in fortified medium. After incubation, retinal vacuolation was reduced.
利用新鲜分离的漂白大鼠视网膜,已经建立了视觉色素再生和光转导恢复的程序。使用含有500微摩尔视网膜异构体混合物(包括9-顺式和11-顺式形式)的磷脂酰胆碱脂质体,并将结果与暗适应视网膜进行比较,暗适应视网膜以类似方式孵育但不含视网膜。在60分钟的孵育中,视紫红质(加异视紫红质)恢复到原始视紫红质浓度的91%,cGMP恢复到87%,饱和时PIII振幅恢复到89%。PIII振幅与对数强度曲线给出的n值在0.6至2.0之间,σ值在85至439量子/微米²之间。死后18至58小时的人视网膜,按上述方法处理后,也产生了光反应。在迄今为止研究的7个视网膜中,视紫红质已再生至0.1-0.35纳摩尔/毫克蛋白质,cGMP再生至23.5-49.2皮摩尔/毫克蛋白质,PIII再生至20-50微伏:在某些情况下还可见b波。n值在0.6至1.0之间变化,σ值在132至3700量子/微米²之间。死后约30小时的视网膜在强化培养基中再孵育24小时后,也可见PIII反应。孵育后,视网膜空泡化减少。