title: Pilot study of the Human Proteome Organisation Brain Proteome Project: applying different 2-DE techniques to monitor proteomic changes during murine brain development.
abstract: The Human Proteome Organisation Brain Proteome Project aims at coordinating neuroproteomic activities with respect to analysis of development, aging, and evolution in human and mice and at analysing normal aging processes as well as neurodegenerative diseases. Our group participated in the mouse pilot study of this project using two different 2-DE systems, to find out the optimal conditions for comprehensive gel-based differential proteome analysis. Besides the assessment of the best methodical conditions the question of "How many biological replicate analyses have to be performed to get reliable statistically validated results?" was addressed. In total 420 differences were detected in all analyses. Both 2-DE methods were found to be suitable for comprehensive differential proteome analysis. Nevertheless, each of the methods showed substantial advantages and disadvantages resulting in the fact that modification of both systems is essential. From our results we can draw the conclusions that for the future optimal quantitative differential gel-based brain proteome analyses the sample preparation has to be slightly changed, the resolution of the first as well as the second dimension has to be advanced, the number of experiments has to be increased and that the 2D-DIGE system should be applied.
authors.affiliation: Medizinisches Proteom-Center, Ruhr-University Bochum, Bochum, Germany. kai.stuehler@ruhr-uni.bochum.de
authors.forename: Kai
authors.initials: K
authors.lastname: Stühler
authors.forename: Kathy
authors.initials: K
authors.lastname: Pfeiffer
authors.forename: Cornelia
authors.initials: C
authors.lastname: Joppich
authors.forename: Christian
authors.initials: C
authors.lastname: Stephan
authors.forename: Klaus
authors.initials: K
authors.lastname: Jung
authors.forename: Michael
authors.initials: M
authors.lastname: Müller
authors.forename: Oliver
authors.initials: O
authors.lastname: Schmidt
authors.forename: Andre
authors.initials: A
authors.lastname: van Hall
authors.forename: Michael
authors.initials: M
authors.lastname: Hamacher
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Urfer
authors.forename: Helmut E
authors.initials: HE
authors.lastname: Meyer
authors.forename: Katrin
authors.initials: K
authors.lastname: Marcus
mesh_terms: D000375:Aging / Q000378:metabolism; D000818:Animals; D001921:Brain / Q000196:embryology / Q000254:growth & development / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D007091:Image Processing, Computer-Assisted; D051379:Mice; D008810:Mice, Inbred C57BL; D010865:Pilot Projects; D020543:Proteome / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D020543:Proteome