Tissue profiling of the mammalian central nervous system using human antibody-based proteomics.
A need exists for mapping the protein profiles in the human brain both during normal and disease conditions. Here we studied 800 antibodies generated toward human proteins as part of a Human Protein Atlas program and investigated their suitability for detailed analysis of various levels of a rat brain using immuno-based methods. In this way, the parallel, rather limited analysis of the human brain, restricted to four brain areas (cerebellum, cerebral cortex, hippocampus, and lateral subventricular zone), could be extended in the rat model to 25 selected areas of the brain. Approximately 100 antibodies (12%) revealed a distinct staining pattern and passed validation of specificity using Western blot analysis. These antibodies were applied to coronal sections of the rat brain at 0.7-mm intervals covering the entire brain. We have now produced detailed protein distribution profiles for these antibodies and acquired over 640 images that form the basis of a publicly available portal of an antibody-based Rodent Brain Protein Atlas database (www.proteinatlas.org/rodentbrain). Because of the systematic selection of target genes, the majority of antibodies included in this database are generated against proteins that have not been studied in the brain before. Furthermore optimized tissue processing and colchicine treatment allow a high quality, more extended annotation and detailed analysis of subcellular distributions and protein dynamics.
Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden. j.mulder@abdn.ac.uk
Jan
J
Mulder
Erik
E
Björling
Kalle
K
Jonasson
Henrik
H
Wernérus
Sophia
S
Hober
Tomas
T
Hökfelt
Mathias
M
Uhlén
D000818:Animals; D000906:Antibodies / Q000378:metabolism*; D000918:Antibody Specificity; D001921:Brain / Q000033:anatomy & histology / Q000378:metabolism; D001931:Brain Mapping / Q000379:methods; D002490:Central Nervous System / Q000737:chemistry*; D003078:Colchicine / Q000378:metabolism; D030562:Databases, Protein; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D051381:Rats; D015203:Reproducibility of Results; D046888:Tissue Array Analysis / Q000379:methods*; D050257:Tubulin Modulators / Q000378:metabolism
D000906:Antibodies; D009419:Nerve Tissue Proteins; D050257:Tubulin Modulators; D003078:Colchicine