Barnhill Alison E, Hecker Laura A, Kohutyuk Oksana, Buss Janice E, Honavar Vasant G, Greenlee Heather West
Interdepartmental Neuroscience Program, Iowa State University, Ames, IA USA.
BMC Res Notes. 2010 Jan 27;3:25. doi: 10.1186/1756-0500-3-25.
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.
视杆光感受器的发生、细胞命运决定和分化过程复杂且受多种因素影响。先前的研究已经定义了一种光感受器分化模型,该模型依赖于假定的光感受器细胞内的内在变化以及细胞外部周围组织的变化。我们采用蛋白质组学方法来鉴定在小鼠视网膜视杆细胞发生和分化过程中动态表达的蛋白质。
使用从E13到P5的六个发育阶段来定义视杆光感受器发生和早期分化过程中视网膜蛋白质表达的变化。视网膜蛋白质通过等电点和分子量进行分离。分析凝胶在发育时间内蛋白质斑点强度的变化。从凝胶中挑选出在E17、P0和P5表达达到峰值的蛋白质斑点进行鉴定。基于它们在视杆光感受器最大发生和分化发育时期的动态表达,挑选出239个斑点进行鉴定。在这239个斑点中,有60个被可靠鉴定并代表单一蛋白质。10种蛋白质由多个斑点代表,表明它们经过了翻译后修饰。在鉴定出的42种独特的动态表达蛋白质中,有16种先前已报道与发育中的视网膜相关。
我们的结果代表了首次对包括产前发育在内的发育中小鼠视网膜进行的蛋白质组学研究。我们在发育中小鼠视网膜中鉴定出26种动态表达的蛋白质,其表达先前未与视网膜发育相关联。