title: Microscopy ambient ionization top-down mass spectrometry reveals developmental patterning.
abstract: There is immense cellular and molecular heterogeneity in biological systems. Here, we demonstrate the utility of integrating an inverted light microscope with an ambient ionization source, nanospray electrospray desorption ionization, attached to a high-resolution mass spectrometer to characterize the molecular composition of mouse spinal cords. We detected a broad range of molecules, including peptides and proteins, as well as metabolites such as lipids, sugars, and other small molecules, including S-adenosyl methionine and glutathione, through top-down MS. Top-down analysis revealed variation in the expression of Hb, including the transition from fetal to adult Hb and heterogeneity in Hb subunits consistent with the genetic diversity of the mouse models. Similarly, temporal changes to actin-sequestering proteins β-thymosins during development were observed. These results demonstrate that interfacing microscopy with ambient ionization provides the means to perform targeted in situ ambient top-down mass spectral analysis to study the pattern of proteins, lipids, and sugars in biologically heterogeneous samples.
authors: Department of Chemistry and Biochemistry, Department of Medicine, and Division of Biological Sciences, University of California, San Diego, La Jolla, CA 92093.
authors: Cheng-Chih
authors: CC
authors: Hsu
authors: Nicholas M
authors: NM
authors: White
authors: Marito
authors: M
authors: Hayashi
authors: Eugene C
authors: EC
authors: Lin
authors: Tiffany
authors: T
authors: Poon
authors: Indroneal
authors: I
authors: Banerjee
authors: Ju
authors: J
authors: Chen
authors: Samuel L
authors: SL
authors: Pfaff
authors: Eduardo R
authors: ER
authors: Macagno
authors: Pieter C
authors: PC
authors: Dorrestein
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019521:Body Patterning; D050260:Carbohydrate Metabolism; D005260:Female; D006454:Hemoglobins / Q000235:genetics / Q000378:metabolism; D050356:Lipid Metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008813:Mice, Inbred ICR; D018345:Mice, Knockout; D008822:Mice, Transgenic; D008853:Microscopy / Q000295:instrumentation / Q000379:methods*; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000378:metabolism; D011247:Pregnancy; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation / Q000379:methods*; D013116:Spinal Cord / Q000196:embryology / Q000254:growth & development* / Q000378:metabolism*; D053719:Tandem Mass Spectrometry / Q000295:instrumentation / Q000379:methods; D013947:Thymosin / Q000235:genetics / Q000378:metabolism
chemical_list: D006454:Hemoglobins; D009419:Nerve Tissue Proteins; D013947:Thymosin
keywords: ambient MS; central nervous system; embryogenesis; light microscopy; top-down MS analysis
references: Anal Chem. 2012 Feb 7;84(3):1517-25
references: 22166099
references: Biochem Genet. 1982 Feb;20(1-2):199-208
references: 7092800
references: Proc Natl Acad Sci U S A. 1992 Jan 1;89(1):286-9
references: 1309610
references: Nature. 2003 Mar 13;422(6928):198-207
references: 12634793
references: Annu Rev Phys Chem. 2013;64:481-505
references: 23331308
references: ACS Nano. 2011 Jul 26;5(7):5526-31
references: 21639403
references: Expert Opin Biol Ther. 2012 Jan;12(1):37-51
references: 22074294
references: Biochim Biophys Acta. 1973 Mar 23;303(1):52-60
references: 4573825
references: Circ Res. 2012 Feb 3;110(3):456-64
references: 22158707
references: J Lipid Res. 2009 Nov;50(11):2290-8
references: 19429885
references: Science. 2006 Apr 14;312(5771):212-7
references: 16614208
references: Anal Chem. 2004 Jun 1;76(11):197A-203A
references: 15190879
references: Nat Rev Genet. 2000 Oct;1(1):20-9
references: 11262869
references: J Neurochem. 1991 Feb;56(2):457-61
references: 1988550
references: Cancer Invest. 2009 Dec;27(10):1016-22
references: 19909017
references: Nature. 1961 May 27;190:818-9
references: 13687338
references: Biochemistry. 2004 Nov 23;43(46):14792-801
references: 15544350
references: Ther Drug Monit. 2005 Dec;27(6):747-51
references: 16404815
references: J Mass Spectrom. 2012 Jan;47(1):29-33
references: 22282086
references: Science. 2013 Jan 11;339(6116):222-6
references: 23118012
references: Cell Motil Cytoskeleton. 2009 Oct;66(10):839-51
references: 19405116
references: Development. 2008 Mar;135(6):1097-106
references: 18272593
references: Endocrinology. 2008 Jul;149(7):3274-8
references: 18403475
references: J Am Chem Soc. 2002 Jan 30;124(4):672-8
references: 11804498
references: Nat Methods. 2007 Oct;4(10):828-33
references: 17901873
references: Vet Pathol. 2005 Jul;42(4):405-26
references: 16006601
references: J Biol Chem. 1993 Jan 5;268(1):502-9
references: 8416954
references: Anal Chem. 1997 Dec 1;69(23):4751-60
references: 9406525
references: Proc Natl Acad Sci U S A. 2008 Nov 25;105(47):18126-31
references: 18776051
references: Anal Bioanal Chem. 2012 Dec;404(10):2915-26
references: 23052870
references: Biochim Biophys Acta. 2013 Feb;1830(2):2354-60
references: 22986149
references: Nucleic Acids Res. 2004 Jul 1;32(Web Server issue):W340-5
references: 15215407
references: Mol Cell Neurosci. 2010 Dec;45(4):335-44
references: 20654717
references: Rapid Commun Mass Spectrom. 2013 Jul 15;27(13):1429-36
references: 23722677
references: Genes Dev. 1999 Jan 1;13(1):112-24
references: 9887104
references: Cell. 2006 Nov 3;127(3):635-48
references: 17081983
references: FEBS J. 2012 Feb;279(3):361-9
references: 22129306
references: Science. 2004 Oct 15;306(5695):471-3
references: 15486296
references: Anal Chem. 1994 Sep 15;66(18):2801-8
references: 7978294
references: Proc Natl Acad Sci U S A. 2012 Jun 26;109(26):E1743-52
references: 22586093
references: J Mol Biol. 1967 Jul 14;27(1):9-16
references: 5340470
references: J Mass Spectrom. 2010 Jul;45(7):703-14
references: 20623627
references: Science. 2006 Oct 6;314(5796):65-6
references: 17023639
references: Anal Bioanal Chem. 2012 Jan;402(2):625-45
references: 21769551
references: Cell. 2012 Jan 20;148(1-2):273-84
references: 22265416
references: J Proteome Res. 2008 Dec;7(12):5314-26
references: 19367708
references: Int J Biochem Cell Biol. 2001 Mar;33(3):205-20
references: 11311852
references: Nature. 1999 Apr 15;398(6728):622-7
references: 10217145
references: Anal Chem. 2012 Jan 3;84(1):141-8
references: 22098105
references: Brain Res Mol Brain Res. 1990 Jul;8(2):129-35
references: 2169566
references: Anal Chem. 2009 Nov 1;81(21):9153-7
references: 19788281