Evaluation of 2‐DE protein patterns from pre‐ and postnatal stages of the mouse brain

Abstract

Brains of the mouse from three developmental stages, embryo day 16 (Ed16), postnatal stage one week (1W) and eight weeks (8W), were distributed to different laboratories for a collaborative proteome analysis (The Human Brain Proteome Project). As one of the laboratories involved in this project, we separated total protein extracts of the brains by large gel 2-DE. From the 2-DE protein patterns a section was evaluated for each of the three stages according to resolution, reproducibility and quantitative changes using an image analysis software. The evaluated pattern section was selected to allow comparisons of 2-DE patterns between different laboratories on the basis of optimum separation. Changes in protein expression were analysed within two phases of development: Stage Ed16 versus stage 1W and stage 1W versus stage 8W. Out of the 200 protein spots evaluated 5-6% showed quantitative changes in the range of > or = 30% between two stages. The relationship in the frequency of up- and down-regulated protein spots differed between the two investigated phases. Most of the protein spots which showed altered expression between two stages were identified by MS. High quality in protein separation and evaluation is demonstrated.

Brief Alteration of NMDA or GABAA Receptor-mediated Neurotransmission Has Long Term Effects on the Developing Cerebral Cortex

“…1). As reported previously by our group, 30 -40% of protein spots in 2-DE gels of whole brain samples change between embryonic day 16 and P7 as well as between P7 and P56, and the number of up-regulated protein isoforms are balanced against the number of down-regulated proteins during postnatal brain development (23). Protein regulation levels may be even higher than can be shown by quantitative measurements of silver-stained gels.…”

FD-LC-MS/MS Method for Determining Protein Expression and Elucidating Biochemical Events in Tissues and Cells

“…For example, only 40 µg of brain segment/injection was used in the study, while hundreds of micrograms of brain segment was required in general brain proteome analysis using 2-DE methods. [26][27][28][29][30] The existence of 28 regionally-specific age-related proteins in mouse brain was demonstrated in this study, seven of which have been reported for the first time. Figure 2 presents a comparison of the peak heights of each of the age-related altered proteins among three aging stages (4, 12 and 20 weeks) in Ctx.…”

Proteomic Studies on the Development of the Central Nervous System and Beyond

“…There have also been other similar HUPO BPP programs [8][9][10][11]. These studies have identified a total of 1,999 spots matched between all gels, among which 206 spots were differentially expressed between the different stages: 122 spots were highest in intensity in embryonic stage E16, 26 were highest in the juvenile group (P7) and 58 spots were highest in the adult stage (P56), revealing a pattern of temporal expression for the examined proteins [107].…”

“…Klose et al also conducted a HUPO BPP pilot study using mouse brains at different developmental stages. They identified about 200 protein spots, most of which showed altered expression between the two developmental stages and were further validated by MS, showing a high quality of protein separation and evaluation [11].…”