title: Analysis of peptides secreted from cultured mouse brain tissue.
abstract: Peptides represent a major class of cell-cell signaling molecules. Most peptidomic studies have focused on peptides present in brain or other tissues. For a peptide to function in intercellular signaling, it must be secreted. The present study was undertaken to identify the major peptides secreted from mouse brain slices that were cultured in oxygenated buffer for 3-4h. Approximately 75% of the peptides identified in extracts of cultured slices matched the previously reported peptide content of heat-inactivated mouse brain tissue, whereas only 2% matched the peptide content of unheated brain tissue; the latter showed a large number of postmortem changes. As found with extracts of heat-inactivated mouse brain, the extracts of cultured brain slices represented secretory pathway peptides as well as peptides derived from intracellular proteins such as those present in the cytosol and mitochondria. A subset of the peptides detected in the extracts of the cultured slices was detected in the culture media. The vast majority of secreted peptides arose from intracellular proteins and not secretory pathway proteins. The peptide RVD-hemopressin, a CB1 cannabinoid receptor agonist, was detected in culture media, which is consistent with a role for RVD-hemopressin as a non-classical neuropeptide. Taken together with previous studies, the present results show that short-term culture of mouse brain slices is an appropriate system to study peptide secretion, especially the non-conventional pathway(s) by which peptides produced from intracellular proteins are secreted. This article is part of a Special Issue entitled: An Updated Secretome.
authors: Department of Neuroscience, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
authors: Julia S
authors: JS
authors: Gelman
authors: Sayani
authors: S
authors: Dasgupta
authors: Iryna
authors: I
authors: Berezniuk
authors: Lloyd D
authors: LD
authors: Fricker
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000378:metabolism*; D001923:Brain Chemistry; D005260:Female; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D009479:Neuropeptides / Q000032:analysis* / Q000378:metabolism*; D040901:Proteomics; D055571:Secretory Pathway; D046509:Tissue Culture Techniques
chemical_list: D009479:Neuropeptides
keywords: 4-trimethylammoniumbutyrate; 4-trimethylammoniumbutyryl-N-hydroxysuccinimide; Bioactive peptide; DMSO; Dimethyl Sulfoxide; LC/MS; MS/MS; Neuropeptide; Peptidomics; TMAB; TMAB-NHS; aCSF; artificial cerebrospinal fluid; liquid chromatography/mass spectrometry; m/z; mass charge ratio; tandem mass spectrometry
references: J Biol Chem. 2008 Apr 18;283(16):10347-56
references: 18272524
references: Annu Rev Anal Chem (Palo Alto Calif). 2008;1:451-83
references: 20636086
references: Mol Cell. 2010 Feb 26;37(4):529-40
references: 20188671
references: Methods Mol Biol. 2011;768:307-23
references: 21805251
references: J Neurochem. 2004 Nov;91(4):769-77
references: 15525330
references: J Proteome Res. 2011 Apr 1;10(4):1583-92
references: 21204522
references: J Neurochem. 1990 Aug;55(2):461-7
references: 2370548
references: J Neurochem. 2010 Jun;113(5):1275-84
references: 20367757
references: J Biol Chem. 2009 May 22;284(21):14105-16
references: 19282285
references: J Biol Chem. 2008 Sep 5;283(36):24448-59
references: 18617518
references: J Proteome Res. 2006 Dec;5(12):3368-75
references: 17137338
references: Traffic. 2011 Jul;12(7):799-805
references: 21585635
references: Prog Drug Res. 2003;61:1-37
references: 14674607
references: J Biochem. 2006 Jul;140(1):7-12
references: 16877763
references: J Neurochem. 2010 Jul;114(1):215-25
references: 20412386
references: Annu Rev Neurosci. 2003;26:105-31
references: 14527267
references: J Biol Chem. 2002 Oct 25;277(43):40368-74
references: 12171930
references: J Comp Neurol. 2009 Aug 10;515(5):538-47
references: 19479992
references: Proteomics. 2012 Aug;12(17):2668-81
references: 22740317
references: Brain Res Rev. 2007 Apr;54(1):34-66
references: 17222914
references: Biopolymers. 2009 Jul;91(7):505-13
references: 19226619
references: Curr Opin Chem Biol. 2009 Jun;13(3):284-90
references: 19553156
references: Mol Cell Proteomics. 2011 Mar;10(3):M900229MCP200
references: 20110281
references: Hum Reprod. 2008 Mar;23(3):635-41
references: 18216035
references: PLoS One. 2013;8(1):e53263
references: 23308178
references: Annu Rev Cell Dev Biol. 2008;24:287-308
references: 18590485
references: J Proteome Res. 2003 Mar-Apr;2(2):213-9
references: 12716136
references: Mass Spectrom Rev. 2006 Jan-Feb;25(1):77-98
references: 15937922
references: AAPS J. 2010 Sep;12(3):279-89
references: 20383670
references: J Am Soc Nephrol. 2008 Aug;19(8):1500-8
references: 18448584
references: ACS Chem Neurosci. 2010 Jan 8;1(4):306-314
references: 20495672
references: Curr Protoc Protein Sci. 2011 Aug;Chapter 18:Unit18.13
references: 21842468
references: Anal Chem. 2006 Oct 15;78(20):7267-72
references: 17037931
references: Mol Cell Proteomics. 2005 Sep;4(9):1391-405
references: 15970582
references: J Proteome Res. 2007 Dec;6(12):4667-76
references: 17979226
references: Proc Natl Acad Sci U S A. 2009 Sep 8;106(36):15454-9
references: 19717439
references: Proc Natl Acad Sci U S A. 2008 Aug 26;105(34):12527-32
references: 18719122
references: Mol Biosyst. 2010 Aug;6(8):1355-65
references: 20428524
references: Eukaryot Cell. 2006 Sep;5(9):1560-70
references: 16963638
references: Proteomics. 2007 Dec;7(24):4445-56
references: 18072205
references: J Neurochem. 2010 May;113(4):871-80
references: 20202081
references: Proc Natl Acad Sci U S A. 2010 Aug 10;107(32):14508-13
references: 20699386
references: Proc Natl Acad Sci U S A. 1999 Jun 8;96(12):6643-7
references: 10359765
references: Biopolymers. 2007;88(2):190-8
references: 17245751
references: Mass Spectrom Rev. 2006 Mar-Apr;25(2):327-44
references: 16404746
references: J Chromatogr B Analyt Technol Biomed Life Sci. 2003 Jul 15;792(1):33-48
references: 12828995
references: Immunity. 2004 Apr;20(4):495-506
references: 15084277
references: Nat Rev Mol Cell Biol. 2002 Feb;3(2):122-31
references: 11836514
references: Anal Chem. 2008 Dec 1;80(23):9298-309
references: 19551992
references: Proteomics. 2012 Aug;12(17):2641-55
references: 22740335
references: J Proteome Res. 2010 Oct 1;9(10):5047-52
references: 20681733
references: Proc Natl Acad Sci U S A. 2007 Dec 18;104(51):20588-93
references: 18077343
references: Methods Mol Biol. 2012;829:487-503
references: 22231834
references: J Biol Chem. 2006 Mar 3;281(9):5668-76
references: 16407281
references: FASEB J. 2009 Sep;23(9):3020-9
references: 19380512
references: Anal Chem. 2007 Jan 1;79(1):15-6, 18-21
references: 17262921
references: Int J Dev Biol. 2008;52(7):873-86
references: 18956317
references: J Neurosci. 2000 Jan 15;20(2):639-48
references: 10632593
references: J Proteome Res. 2012 Mar 2;11(3):1981-90
references: 22304392
references: FASEB J. 2010 Jun;24(6):1813-23
references: 20061535
references: Trends Mol Med. 2005 Sep;11(9):421-9
references: 16099219
references: Neuropharmacology. 2004;47 Suppl 1:345-58
references: 15464149
references: Trends Endocrinol Metab. 2009 Jan;20(1):25-33
references: 19056296
references: Science. 2010 Jul 16;329(5989):336-9
references: 20647469
references: J Neurochem. 2010 Mar;112(5):1168-79
references: 19968759
references: Comb Chem High Throughput Screen. 2001 Apr;4(2):207-17
references: 11281836