Ortín-Martínez Arturo, Salinas-Navarro Manuel, Nadal-Nicolás Francisco Manuel, Jiménez-López Manuel, Valiente-Soriano Francisco Javier, García-Ayuso Diego, Bernal-Garro José Manuel, Avilés-Trigueros Marcelino, Agudo-Barriuso Marta, Villegas-Pérez María Paz, Vidal-Sanz Manuel
Departamento de Oftalmología, Facultad de Medicina, Universidad de Murcia, and Instituto Murciano de Investigación Biosanitaria Virgen de la Arrixaca (IMIB-Arrixaca), 30100 Murcia, Spain.
Departamento de Oftalmología, Facultad de Medicina, Universidad de Murcia, and Instituto Murciano de Investigación Biosanitaria Virgen de la Arrixaca (IMIB-Arrixaca), 30100 Murcia, Spain.
Exp Eye Res. 2015 Mar;132:17-33. doi: 10.1016/j.exer.2015.01.006. Epub 2015 Jan 7.
To investigate the long-term effects of laser-photocoagulation (LP)-induced ocular hypertension (OHT) in the innermost and outermost (outer-nuclear and outer segment)-retinal layers (ORL). OHT was induced in the left eye of adult rats. To investigate the ganglion cell layer (GCL) wholemounts were examined at 1, 3 or 6 months using Brn3a-immunodetection to identify retinal ganglion cells (RGCs) and DAPI-staining to detect all nuclei in this layer. To study the effects of LP on the ORL up to 6 months, retinas were: i) fresh extracted to quantify the levels of rod-, S- and L-opsin; ii) cut in cross-sections for morphometric analysis, or; iii) prepared as wholemounts to quantify and study retinal distributions of entire populations of RGCs (retrogradely labeled with fluorogold, FG), S- and L-cones (immunolabeled). OHT resulted in wedge-like sectors with their apex on the optic disc devoid of Brn3a(+)RGCs but with large numbers of DAPI(+)nuclei. The levels of all opsins diminished by 2 weeks and further decreased to 20% of basal-levels by 3 months. Cross-sections revealed focal areas of ORL degeneration. RGC survival at 15 days represented approximately 28% and did not change with time, whereas the S- and L-cone populations diminished to 65% and 80%, or to 20 and 35% at 1 or 6 months, respectively. In conclusion, LP induces in the GCL selective RGCs loss that does not progress after 1 month, and S- and L-cone loss that progresses for up to 6 months. Thus, OHT results in severe damage to both the innermost and the ORL.
研究激光光凝(LP)诱导的眼内高压(OHT)对视网膜最内层和最外层(外核层和外节层)视网膜层(ORL)的长期影响。在成年大鼠的左眼诱导产生OHT。为了研究神经节细胞层(GCL),在1、3或6个月时使用Brn3a免疫检测来识别视网膜神经节细胞(RGCs),并用DAPI染色来检测该层中的所有细胞核。为了研究LP对ORL长达6个月的影响,视网膜被处理如下:i)新鲜提取以定量视杆、S和L视蛋白的水平;ii)切成横截面进行形态计量分析,或iii)制成整装片以定量和研究RGCs(用荧光金逆行标记,FG)、S和L锥体(免疫标记)的整个群体的视网膜分布。OHT导致楔形区域,其顶点位于视盘,缺乏Brn3a(+)RGCs,但有大量DAPI(+)细胞核。所有视蛋白水平在2周时下降,到3个月时进一步降至基础水平的20%。横截面显示ORL变性的局灶区域。15天时RGC存活率约为28%,且随时间无变化,而S和L锥体群体在1个月或6个月时分别降至65%和80%,或降至20%和35%。总之,LP诱导GCL中选择性RGCs丢失,1个月后不再进展,以及S和L锥体丢失,持续长达6个月。因此,OHT导致视网膜最内层和ORL均受到严重损伤。