Characterization of the retinal proteome during rod photoreceptor genesis.
Characterization of the retinal proteome during rod photoreceptor genesis.
复制标题
DOI：
10.1186/1756-0500-3-25
复制
发表时间：
2010-01-27
影响因子：
1.8
通讯作者：
Greenlee HW
中科院分区：
文献类型：
--
作者：
Barnhill AE;Hecker LA;Kohutyuk O;Buss JE;Honavar VG;Greenlee HW
视杆细胞的发生、细胞命运的决定和分化是一个复杂的多因素过程。以前的研究已经定义了感光细胞分化的模型，该模型依赖于假定感光细胞内的内在变化以及细胞外部的周围组织的变化。我们已经使用了蛋白质组学的方法，以确定蛋白质的动态表达在小鼠视网膜杆的发生和分化。从E13到P5的一系列六个发育年龄被用来定义视杆细胞发生和早期分化过程中视网膜蛋白表达的变化。用等电聚焦点法和分子量法分离视网膜蛋白。分析凝胶在整个发育时间内蛋白质斑点强度的变化。从凝胶中挑取在E17、P0和P5表达达到峰值的蛋白质点用于鉴定。根据其在视杆细胞最大发生和分化发育期的动态表达，筛选出239个位点进行鉴定。在239个点中，其中60个被可靠地鉴定并代表单一蛋白质。10个蛋白质由多个斑点表示，表明它们是后修饰的。在鉴定的42种独特的动态表达蛋白中，有16种先前已报道与发育中的视网膜相关。我们的研究结果代表了第一个蛋白质组学研究的发展小鼠视网膜，包括产前发展。我们确定了26个动态表达的蛋白质在发育中的小鼠视网膜，其表达以前没有与视网膜发育。
The process of rod photoreceptor genesis, cell fate determination and differentiation is complex and multi-factorial. Previous studies have defined a model of photoreceptor differentiation that relies on intrinsic changes within the presumptive photoreceptor cells as well as changes in surrounding tissue that are extrinsic to the cell. We have used a proteomics approach to identify proteins that are dynamically expressed in the mouse retina during rod genesis and differentiation. A series of six developmental ages from E13 to P5 were used to define changes in retinal protein expression during rod photoreceptor genesis and early differentiation. Retinal proteins were separated by isoelectric focus point and molecular weight. Gels were analyzed for changes in protein spot intensity across developmental time. Protein spots that peaked in expression at E17, P0 and P5 were picked from gels for identification. There were 239 spots that were picked for identification based on their dynamic expression during the developmental period of maximal rod photoreceptor genesis and differentiation. Of the 239 spots, 60 of them were reliably identified and represented a single protein. Ten proteins were represented by multiple spots, suggesting they were post-translationally modified. Of the 42 unique dynamically expressed proteins identified, 16 had been previously reported to be associated with the developing retina. Our results represent the first proteomics study of the developing mouse retina that includes prenatal development. We identified 26 dynamically expressed proteins in the developing mouse retina whose expression had not been previously associated with retinal development.