Human post-mortem synapse proteome integrity screening for proteomic studies of postsynaptic complexes
Por:
Bayes, A, Collins, MO, Galtrey, CM, Simonnet, C, Roy, M, Croning, MDR, Gou, G, van de Lagemaat, LN, Milward, D, Whittle, IR, Smith, C, Choudhary, JS, Grant, SGN
Publicada:
28 nov 2014
Resumen:
Background: Synapses are fundamental components of brain circuits and are disrupted in over 100 neurological and psychiatric diseases. The synapse proteome is physically organized into multiprotein complexes and polygenic mutations converge on postsynaptic complexes in schizophrenia, autism and intellectual disability. Directly characterising human synapses and their multiprotein complexes from post-mortem tissue is essential to understanding disease mechanisms. However, multiprotein complexes have not been directly isolated from human synapses and the feasibility of their isolation from post-mortem tissue is unknown.
Results: Here we establish a screening assay and criteria to identify post-mortem brain samples containing well-preserved synapse proteomes, revealing that neocortex samples are best preserved. We also develop a rapid method for the isolation of synapse proteomes from human brain, allowing large numbers of post-mortem samples to be processed in a short time frame. We perform the first purification and proteomic mass spectrometry analysis of MAGUK Associated Signalling Complexes (MASC) from neurosurgical and post-mortem tissue and find genetic evidence for their involvement in over seventy human brain diseases.
Conclusions: We have demonstrated that synaptic proteome integrity can be rapidly assessed from human post-mortem brain samples prior to its analysis with sophisticated proteomic methods. We have also shown that proteomics of synapse multiprotein complexes from well preserved post-mortem tissue is possible, obtaining structures highly similar to those isolated from biopsy tissue. Finally we have shown that MASC from human synapses are involved with over seventy brain disorders. These findings should have wide application in understanding the synaptic basis of psychiatric and other mental disorders.
Filiaciones:
Bayes, A:
Biomed Res Inst Sant Pau, IIB Sant Pau, Mol Physiol Synapse Lab, Barcelona 08025, Spain
Univ Autonoma Barcelona, Bellaterra 08193, Cerdanyola Del, Spain
Collins, MO:
Univ Sheffield, Dept Biomed Sci, Western Bank, Ctr Membrane Interact & Dynam, Sheffield S10 2TN, S Yorkshire, England
Galtrey, CM:
St George Hosp, Dept Neurol, London, England
Simonnet, C:
Univ Edinburgh, Ctr Clin Brain Sci, Genes Cognit Programme, Edinburgh EH16 4SB, Midlothian, Scotland
Univ Edinburgh, Ctr Neuroregenerat, Edinburgh EH16 4SB, Midlothian, Scotland
Roy, M:
Univ Edinburgh, Ctr Clin Brain Sci, Genes Cognit Programme, Edinburgh EH16 4SB, Midlothian, Scotland
Univ Edinburgh, Ctr Neuroregenerat, Edinburgh EH16 4SB, Midlothian, Scotland
Croning, MDR:
Univ Edinburgh, Ctr Clin Brain Sci, Genes Cognit Programme, Edinburgh EH16 4SB, Midlothian, Scotland
Univ Edinburgh, Ctr Neuroregenerat, Edinburgh EH16 4SB, Midlothian, Scotland
Gou, G:
Biomed Res Inst Sant Pau, IIB Sant Pau, Mol Physiol Synapse Lab, Barcelona 08025, Spain
Univ Autonoma Barcelona, Bellaterra 08193, Cerdanyola Del, Spain
van de Lagemaat, LN:
Univ Edinburgh, Ctr Clin Brain Sci, Genes Cognit Programme, Edinburgh EH16 4SB, Midlothian, Scotland
Univ Edinburgh, Ctr Neuroregenerat, Edinburgh EH16 4SB, Midlothian, Scotland
Milward, D:
Linguamatics, Cambridge CB4 0WG, England
Whittle, IR:
Univ Edinburgh, Ctr Clin Brain Sci, Genes Cognit Programme, Edinburgh EH16 4SB, Midlothian, Scotland
Univ Edinburgh, Ctr Neuroregenerat, Edinburgh EH16 4SB, Midlothian, Scotland
Smith, C:
Univ Edinburgh, Ctr Clin Brain Sci, Acad Dept Neuropathol, Edinburgh EH16 4SB, Midlothian, Scotland
Choudhary, JS:
Wellcome Trust Sanger Inst, Hinxton, Cambs, England
Grant, SGN:
Univ Edinburgh, Ctr Clin Brain Sci, Genes Cognit Programme, Edinburgh EH16 4SB, Midlothian, Scotland
Univ Edinburgh, Ctr Neuroregenerat, Edinburgh EH16 4SB, Midlothian, Scotland
Gold, Green Published, Green Accepted
|