AIM： To identify the underlying cellular and molecular changes in retinitis pigmentosa （RP）. METHODS： Label-free quantification-based proteomics analysis, with its advantages of being more economic and consisting...AIM： To identify the underlying cellular and molecular changes in retinitis pigmentosa （RP）. METHODS： Label-free quantification-based proteomics analysis, with its advantages of being more economic and consisting of simpler procedures, has been used with increasing frequency in modern biological research. Dystrophic RCS rats, the first laboratory animal model for the study of RP, possess a similar pathological course as human beings with the diseases. Thus, we employed a comparative proteomics analysis approach for in-depth proteome profiling of retinas from dystrophic RCS rats and non-dystrophic congenic controls through Linear Trap Quadrupole - orbitrap MS/MS, to identify the significant differentially expressed proteins （DEPs）. Bioinformatics analyses, including Gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway annotation and upstream regulatory analysis, were then performed on these retina proteins. Finally, a Western blotting experiment was carried out to verify the difference in the abundance of transcript factor E2F1. RESULTS： In this study, we identified a total of 2375 protein groups from the retinal protein samples of RCS rats and non-dystrophic congenic controls. Four hundred thirty-four significantly DEPs were selected by Student＇s t-test. Based on the results of the bioinformatics analysis, we identified mitochondrial dysfunction and transcription factor E2F1 as the key initiation factors in early retinal degenerative process. CONCLUSION： We showed that the mitochondrial dysfunction and the transcription factor E2F1 substantially contribute to the disease etiology of RP. The results provide a new potential therapeutic approach for this retinal degenerative disease.展开更多
目的:通过下丘脑弓状核乙酰胆碱表达的检测、图像处理、图像分析,应用阳性神经元面积百分比(APPN)和染色灰度相对强度值(RISGL)相结合的方法,探讨免疫组化(IHC)图像定量分析的特点。方法:样本来源于运动性免疫抑制过程中SD大鼠下丘脑弓...目的:通过下丘脑弓状核乙酰胆碱表达的检测、图像处理、图像分析,应用阳性神经元面积百分比(APPN)和染色灰度相对强度值(RISGL)相结合的方法,探讨免疫组化(IHC)图像定量分析的特点。方法:样本来源于运动性免疫抑制过程中SD大鼠下丘脑弓状核胆碱能阳性神经元乙酰胆碱表达的免疫组化切片,包括第0、2、4、6周与每周Control、Immediately after exercise、3 hours after exercise三组共计12组(n=6)。采用免疫组化技术测定其乙酰胆碱(ACh)的表达,并根据ACh阳性神经元总面积(TAPN)、阳性神经元灰度平均强度值(AISGL)、APPN、RISGL、APPN/RISGL参数进行免疫组化图像的定量分析,从而比较APPN和RISGL在定量分析中与传统参数的不同及其优势。结果:APPN与TAPN的变化呈现几乎一致的变化特征,能发现相应差异的显著性意义(P<0.05),但APPN的敏感性和抗干扰性更强。RISGL与AISGL呈现的结果并不完全一致,而且APPN结合RISGL比单参数更能反映阳性表达情况。结论:APPN和RISGL免疫组化图像分析参数能较可靠地、准确地进行图像定量分析,APPN与RISGL的结合不仅可反映阳性表达的数量情况,还可以帮助分析其作用机理,优于传统分析参数。展开更多
基金Supported by the National Nature Science Foundation of China(No.81130017)National Basic Research Program of China(973 Program,No.2013CB967002)
文摘AIM： To identify the underlying cellular and molecular changes in retinitis pigmentosa （RP）. METHODS： Label-free quantification-based proteomics analysis, with its advantages of being more economic and consisting of simpler procedures, has been used with increasing frequency in modern biological research. Dystrophic RCS rats, the first laboratory animal model for the study of RP, possess a similar pathological course as human beings with the diseases. Thus, we employed a comparative proteomics analysis approach for in-depth proteome profiling of retinas from dystrophic RCS rats and non-dystrophic congenic controls through Linear Trap Quadrupole - orbitrap MS/MS, to identify the significant differentially expressed proteins （DEPs）. Bioinformatics analyses, including Gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway annotation and upstream regulatory analysis, were then performed on these retina proteins. Finally, a Western blotting experiment was carried out to verify the difference in the abundance of transcript factor E2F1. RESULTS： In this study, we identified a total of 2375 protein groups from the retinal protein samples of RCS rats and non-dystrophic congenic controls. Four hundred thirty-four significantly DEPs were selected by Student＇s t-test. Based on the results of the bioinformatics analysis, we identified mitochondrial dysfunction and transcription factor E2F1 as the key initiation factors in early retinal degenerative process. CONCLUSION： We showed that the mitochondrial dysfunction and the transcription factor E2F1 substantially contribute to the disease etiology of RP. The results provide a new potential therapeutic approach for this retinal degenerative disease.
文摘目的:通过下丘脑弓状核乙酰胆碱表达的检测、图像处理、图像分析,应用阳性神经元面积百分比(APPN)和染色灰度相对强度值(RISGL)相结合的方法,探讨免疫组化(IHC)图像定量分析的特点。方法:样本来源于运动性免疫抑制过程中SD大鼠下丘脑弓状核胆碱能阳性神经元乙酰胆碱表达的免疫组化切片,包括第0、2、4、6周与每周Control、Immediately after exercise、3 hours after exercise三组共计12组(n=6)。采用免疫组化技术测定其乙酰胆碱(ACh)的表达,并根据ACh阳性神经元总面积(TAPN)、阳性神经元灰度平均强度值(AISGL)、APPN、RISGL、APPN/RISGL参数进行免疫组化图像的定量分析,从而比较APPN和RISGL在定量分析中与传统参数的不同及其优势。结果:APPN与TAPN的变化呈现几乎一致的变化特征,能发现相应差异的显著性意义(P<0.05),但APPN的敏感性和抗干扰性更强。RISGL与AISGL呈现的结果并不完全一致,而且APPN结合RISGL比单参数更能反映阳性表达情况。结论:APPN和RISGL免疫组化图像分析参数能较可靠地、准确地进行图像定量分析,APPN与RISGL的结合不仅可反映阳性表达的数量情况,还可以帮助分析其作用机理,优于传统分析参数。
