G B Bietti Foundation, Istituto di Ricovero e Cura a Carattere Scientifico, 00198 Rome, Italy.
J Neurosci. 2012 Sep 19;32(38):13010-21. doi: 10.1523/JNEUROSCI.1913-12.2012.
Loss of retinoschisin (RS1) in Rs1 knock-out (Rs1-KO) retina produces a post-photoreceptor phenotype similar to X-linked retinoschisis in young males. However, Rs1 is expressed strongly in photoreceptors, and Rs1-KO mice have early reduction in the electroretinogram a-wave. We examined light-activated transducin and arrestin translocation in young Rs1-KO mice as a marker for functional abnormalities in maturing rod photoreceptors. We found a progressive reduction in luminance threshold for transducin translocation in wild-type (WT) retinas between postnatal days P18 and P60. At P21, the threshold in Rs1-KO retinas was 10-fold higher than WT, but it decreased to <2.5-fold higher by P60. Light-activated arrestin translocation and re-translocation of transducin in the dark were not affected. Rs1-KO rod outer segment (ROS) length was significantly shorter than WT at P21 but was comparable with WT at P60. These findings suggested a delay in the structural and functional maturation of Rs1-KO ROS. Consistent with this, transcription factors CRX and NRL, which are fundamental to maturation of rod protein expression, were reduced in ROS of Rs1-KO mice at P21 but not at P60. Expression of transducin was 15-30% lower in P21 Rs1-KO ROS and transducin GTPase hydrolysis was nearly twofold faster, reflecting a 1.7- to 2.5-fold increase in RGS9 (regulator of G-protein signaling) level. Transduction protein expression and activity levels were similar to WT at P60. Transducin translocation threshold elevation indicates photoreceptor functional abnormalities in young Rs1-KO mice. Rapid reduction in threshold coupled with age-related changes in transduction protein levels and transcription factor expression are consistent with delayed maturation of Rs1-KO photoreceptors.
视蛋白缺失(RS1)在 Rs1 敲除(Rs1-KO)视网膜中产生类似于年轻男性中 X 连锁性视网膜劈裂的后光感受器表型。然而,RS1 在光感受器中表达强烈,并且 Rs1-KO 小鼠的视网膜电图 a 波早期减少。我们检查了年轻的 Rs1-KO 小鼠中的光激活转导蛋白和阻滞蛋白转位,作为成熟杆状光感受器中功能异常的标志物。我们发现,在 WT 视网膜中,从出生后第 18 天到第 60 天,转导蛋白转位的亮度阈值呈进行性降低。在 P21 时,Rs1-KO 视网膜的阈值比 WT 高 10 倍,但到 P60 时降至<2.5 倍。光激活阻滞蛋白转位和转导蛋白在黑暗中的再转位不受影响。在 P21 时,Rs1-KO 杆状光感受器外节(ROS)长度明显短于 WT,但在 P60 时与 WT 相当。这些发现表明 Rs1-KO ROS 的结构和功能成熟延迟。与此一致的是,对于杆状蛋白表达成熟至关重要的转录因子 CRX 和 NRL,在 P21 时 Rs1-KO 小鼠的 ROS 中减少,但在 P60 时没有减少。在 P21 Rs1-KO ROS 中,转导蛋白的表达降低了 15-30%,并且转导蛋白 GTP 水解快了近两倍,反映出 RGS9(G 蛋白信号转导调节剂)水平增加了 1.7-2.5 倍。在 P60 时,转导蛋白表达和活性水平与 WT 相似。转导蛋白转位阈值升高表明年轻的 Rs1-KO 小鼠中光感受器功能异常。阈值的快速降低与转导蛋白水平和转录因子表达的年龄相关变化一致,表明 Rs1-KO 光感受器的成熟延迟。