Lyubarsky Arkady L, Daniele Lauren L, Pugh Edward N
Department of Ophthalmology, University of Pennsylvania, F.M. Kirby Center for Molecular Ophthalmology, Stellar-Chance Building, Room 309B, 422 Curie Boulevard, Philadelphia, PA 19104-6069, USA.
Vision Res. 2004 Dec;44(28):3235-51. doi: 10.1016/j.visres.2004.09.019.
To quantify the rate at which light in a ganzfeld produces photoisomerizations in mouse rods in situ, we measured the rate of rhodopsin bleaching in eyes of recently euthanized mice with fully dilated pupils. The amount of rhodopsin declined as a first-order (exponential) function of the duration of the exposure at the luminance of 920 scot cd m(-2): the rate constants of bleaching were 8.3 x 10(-6) and 2.8 x 10(-5) s(-1) (scot cd(-1)m2)(-1) for C57B1/6 and 129P3/J mice, respectively. When the approximately 3-fold difference in effective areas of the pupils of the mice are taken into consideration, the bleaching rates for both strains become essentially the same, 2.6 x 10(-6) fraction rhodopsin (scot Td s)(-1). Assuming 7 x 10(7) rhodopsin molecules per rod, this bleaching rate yields the result that a flash of 1 scot Td s produces 181 photoisomerizations per rod, a value close to that derived from analysis of the collecting area of the rod for axially propagating light. We measured the electroretinograms of mice of the two strains reared under controlled illumination conditions (2 and 100 lux), and compared their properties, using the calibrations to determine the absolute sensitivities of the b-wave and a-waves. The intensity that produces a half-saturating rod b-wave response is 0.3-0.6 photoisomerizations rod(-1), and the amplification constant of the rod a-wave is 5-6 s(-2) photoisomerization(-1), with little dependence on the strain.
为了量化全视野中的光在原位小鼠视杆细胞中产生光异构化的速率,我们测量了近期安乐死且瞳孔充分扩张的小鼠眼睛中视紫红质的漂白速率。视紫红质的量随着在920 scot cd m(-2)亮度下曝光持续时间的一阶(指数)函数而下降:C57B1/6和129P3/J小鼠的漂白速率常数分别为8.3×10(-6)和2.8×10(-5) s(-1) (scot cd(-1)m2)(-1)。当考虑到小鼠瞳孔有效面积约3倍的差异时,两种品系的漂白速率基本相同,为2.6×10(-6)视紫红质分数 (scot Td s)(-1)。假设每个视杆细胞有7×10(7)个视紫红质分子,这个漂白速率得出的结果是,1 scot Td s的闪光在每个视杆细胞中产生181次光异构化,该值与通过对视杆细胞轴向传播光的收集面积分析得出的值相近。我们测量了在受控光照条件(2和100勒克斯)下饲养的两种品系小鼠的视网膜电图,并比较了它们的特性,使用校准来确定b波和a波的绝对灵敏度。产生半饱和视杆细胞b波反应的强度为0.3至-0.6光异构化/视杆细胞(-1),视杆细胞a波的放大常数为5至6 s(-2)光异构化(-1),对品系的依赖性很小。