目的:检测分析多重剪接RNA结合蛋白2(RNA binding protein with multiple splicing 2,Rbpms2)在小鼠卵母细胞及早期胚胎中的表达及分布情况。方法:运用生物信息学资源分析Rbpms2基因表达的组织分布情况以及Rbpms2蛋白的功能结构域;通过...目的:检测分析多重剪接RNA结合蛋白2(RNA binding protein with multiple splicing 2,Rbpms2)在小鼠卵母细胞及早期胚胎中的表达及分布情况。方法:运用生物信息学资源分析Rbpms2基因表达的组织分布情况以及Rbpms2蛋白的功能结构域;通过免疫印迹法、免疫组织化学及免疫荧光检测,分析Rbpms2在小鼠卵母细胞及早期胚胎中的表达和分布。结果:生物信息学检索显示Rbpms2 mRNA在卵细胞及受精卵中高表达,其蛋白具有一个RNA识别结构域。免疫印记法证实Rbpms2蛋白在小鼠卵母细胞中高表达,同时免疫组织化学显示阳性信号主要集中在卵母细胞胞质中,免疫荧光显示在早期胚胎中,Rbpms2的信号集中在细胞核中。结论:Rbpms2在小鼠卵母细胞及早期胚胎中呈高水平表达,提示其可能在卵母细胞和早期胚胎的发育过程中发挥重要作用。展开更多
Univocal identification of retinal ganglion cells(RGCs) is an essential prerequisite for studying their degeneration and neuroprotection. Before the advent of phenotypic markers, RGCs were normally identified using re...Univocal identification of retinal ganglion cells(RGCs) is an essential prerequisite for studying their degeneration and neuroprotection. Before the advent of phenotypic markers, RGCs were normally identified using retrograde tracing of retinorecipient areas. This is an invasive technique, and its use is precluded in higher mammals such as monkeys. In the past decade, several RGC markers have been described. Here, we reviewed and analyzed the specificity of nine markers used to identify all or most RGCs, i.e., pan-RGC markers, in rats, mice, and macaques. The best markers in the three species in terms of specificity, proportion of RGCs labeled, and indicators of viability were BRN3A, expressed by vision-forming RGCs, and RBPMS, expressed by vision-and non-vision-forming RGCs. NEUN, often used to identify RGCs, was expressed by non-RGCs in the ganglion cell layer, and therefore was not RGC-specific. γ-SYN, TUJ1, and NF-L labeled the RGC axons, which impaired the detection of their somas in the central retina but would be good for studying RGC morphology. In rats, TUJ1 and NF-L were also expressed by non-RGCs. BM88, ERRβ,and PGP9.5 are rarely used as markers, but they identified most RGCs in the rats and macaques and ERRβ in mice. However, PGP9.5 was also expressed by non-RGCs in rats and macaques and BM88 and ERRβ were not suitable markers of viability.展开更多
文摘目的:检测分析多重剪接RNA结合蛋白2(RNA binding protein with multiple splicing 2,Rbpms2)在小鼠卵母细胞及早期胚胎中的表达及分布情况。方法:运用生物信息学资源分析Rbpms2基因表达的组织分布情况以及Rbpms2蛋白的功能结构域;通过免疫印迹法、免疫组织化学及免疫荧光检测,分析Rbpms2在小鼠卵母细胞及早期胚胎中的表达和分布。结果:生物信息学检索显示Rbpms2 mRNA在卵细胞及受精卵中高表达,其蛋白具有一个RNA识别结构域。免疫印记法证实Rbpms2蛋白在小鼠卵母细胞中高表达,同时免疫组织化学显示阳性信号主要集中在卵母细胞胞质中,免疫荧光显示在早期胚胎中,Rbpms2的信号集中在细胞核中。结论:Rbpms2在小鼠卵母细胞及早期胚胎中呈高水平表达,提示其可能在卵母细胞和早期胚胎的发育过程中发挥重要作用。
基金supported by the Spanish Ministry of Economy and Competitiveness(PID2019-106498GB-I0)Instituto de Salud Carlos III,Fondo Europeo de Desarrollo Regional“Una manera de hacer Europa”(PI19/00071)+2 种基金Fundación Séneca,Agencia de Ciencia y Tecnología Región de Murcia(19881/GERM/15)Spanish Ministry of Science and Innovation(PID 2019-106498 GB-I00)Intramural Research Program of the National Eye Institute,National Institutes of Health(NIH/NEI RO1 EY029087)。
文摘Univocal identification of retinal ganglion cells(RGCs) is an essential prerequisite for studying their degeneration and neuroprotection. Before the advent of phenotypic markers, RGCs were normally identified using retrograde tracing of retinorecipient areas. This is an invasive technique, and its use is precluded in higher mammals such as monkeys. In the past decade, several RGC markers have been described. Here, we reviewed and analyzed the specificity of nine markers used to identify all or most RGCs, i.e., pan-RGC markers, in rats, mice, and macaques. The best markers in the three species in terms of specificity, proportion of RGCs labeled, and indicators of viability were BRN3A, expressed by vision-forming RGCs, and RBPMS, expressed by vision-and non-vision-forming RGCs. NEUN, often used to identify RGCs, was expressed by non-RGCs in the ganglion cell layer, and therefore was not RGC-specific. γ-SYN, TUJ1, and NF-L labeled the RGC axons, which impaired the detection of their somas in the central retina but would be good for studying RGC morphology. In rats, TUJ1 and NF-L were also expressed by non-RGCs. BM88, ERRβ,and PGP9.5 are rarely used as markers, but they identified most RGCs in the rats and macaques and ERRβ in mice. However, PGP9.5 was also expressed by non-RGCs in rats and macaques and BM88 and ERRβ were not suitable markers of viability.
