The mobilization of nutrient reserves into the ovaries of Aedes aegypti mosquitoes after sugar-feeding plays a vital role in the female's reproductive maturation. In the present work, three-dimensional secondary ion mass spectrometry imaging (3D-SIMS) was used to generate ultrahigh spatial resolution (~1 μm) chemical maps and study the composition and spatial distribution of lipids at the single ovarian follicle level (~100 μm in size). 3D-Mass Spectrometry Imaging (3D-MSI) allowed the identification of cellular types in the follicle (oocyte, nurse and follicular cells) using endogenous markers, and revealed that most of the triacyglycerides (TGs) were compartmentalized in the oocyte region. By comparing follicles from water-fed and sugar-fed females (n=2), 3D-MSI-Time of Flight-SIMS showed that TGs were more abundant in ovarian follicles of sugar-fed females; despite relative sample reproducibility per feeding condition, more biological replicates will better support the trends observed. While the current 3D-MSI-TOF-SIMS does not permit MS/MS analysis of the lipid species, complementary LC-MS/MS analysis of the ovarian follicles aided tentative lipid assignments of the SIMS data. The combination of these MS approaches is giving us a first glimpse of the distribution of functionally relevant ovarian lipid molecules at the cellular level. These new tools can be used to investigate the roles of different lipids on follicle fitness and overall mosquito reproductive output.

Biomolecular Sciences Institute Florida International University Miami Florida 33199 United States

Department of Biological Sciences Florida International University Miami Florida 33199 United States

Department of Chemistry and Biochemistry Florida International University Miami Florida 33199 United States

Institute of Parasitology Biology Centre CAS Ceske Budejovice Czech Republic; and

Zobrazit více v PubMed

Hu T, Zhang J-L, Mass-spectrometry-based lipidomics, Journal of Separation Science, 41 (2017) 351–372. PubMed

Rustam YH, Reid GE, Analytical Challenges and Recent Advances in Mass Spectrometry Based Lipidomics, Analytical Chemistry, 90 (2018) 374–397. PubMed

Bandu R, Mok HJ, Kim KP, Phospholipids as cancer biomarkers: Mass spectrometry-based analysis, Mass Spectrometry Reviews, 37 (2016) 107–138. PubMed

Ulmer CZ, Yost RA, Chen J, Mathews CE, Garrett TJ, Liquid chromatography-mass spectrometry metabolic and lipidomic sample preparation workflow for suspension-cultured mammalian cells using Jurkat T lymphocyte cells, J. Proteomics Bioinf, 8 (2015) 360/361–360/367. PubMed PMC

Veličković D, Chu RK, Carrell AA, Thomas M, Paša-Tolić L, Weston DJ, Anderton CR, Multimodal MSI in Conjunction with Broad Coverage Spatially Resolved MS2 Increases Confidence in Both Molecular Identification and Localization, Analytical Chemistry, 90 (2018) 702–707. PubMed

Van Berkel GJ, Kertesz V, Koeplinger KA, Vavrek M, Kong A-NT, Liquid microjunction surface sampling probe electrospray mass spectrometry for detection of drugs and metabolites in thin tissue sections, Journal of Mass Spectrometry, 43 (2007) 500–508. PubMed

Takáts Z, Wiseman JM, Gologan B, Cooks RG, Mass Spectrometry Sampling Under Ambient Conditions with Desorption Electrospray Ionization, Science, 306 (2004) 471. PubMed

Cooks RG, Ouyang Z, Takats Z, Wiseman JM, Ambient Mass Spectrometry, Science, 311 (2006) 1566. PubMed

Van Berkel GJ, Sanchez AD, Quirke JME, Thin-Layer Chromatography and Electrospray Mass Spectrometry Coupled Using a Surface Sampling Probe, Analytical Chemistry, 74 (2002) 6216–6223. PubMed

Campbell DI, Ferreira CR, Eberlin LS, Cooks RG, Improved spatial resolution in the imaging of biological tissue using desorption electrospray ionization, Analytical and Bioanalytical Chemistry, 404 (2012) 389–398. PubMed

Lamont L, Eijkel GB, Jones EA, Flinders B, Ellis SR, Porta T, Heeren RMA, Vreeken RJ, Targeted Drug and Metabolite Imaging: Desorption Electrospray Ionization combined with Triple Quadrupole Mass Spectrometry, Analytical Chemistry, DOI 10.1021/acs.analchem.8b03857(2018). PubMed DOI PMC

Yin R, Kyle J, Burnum-Johnson K, Bloodsworth KJ, Sussel L, Ansong C, Laskin J, High Spatial Resolution Imaging of Mouse Pancreatic Islets Using Nanospray Desorption Electrospray Ionization Mass Spectrometry, Analytical Chemistry, 90 (2018) 6548–6555. PubMed PMC

Zavalin A, Yang J, Hayden K, Vestal M, Caprioli RM, Tissue protein imaging at 1 μm laser spot diameter for high spatial resolution and high imaging speed using transmission geometry MALDI TOF MS, Analytical and Bioanalytical Chemistry, 407 (2015) 2337–2342. PubMed PMC

Karas M, Hillenkamp F, Laser desorption ionization of proteins with molecular masses exceeding 10,000 daltons, Analytical Chemistry, 60 (1988) 2299–2301. PubMed

Caprioli RM, Farmer TB, Gile J, Molecular Imaging of Biological Samples: Localization of Peptides and Proteins Using MALDI-TOF MS, Anal. Chem, 69 (1997) 4751–4760. PubMed

Baker TC, Han J, Borchers CH, Recent advancements in matrix-assisted laser desorption/ionization mass spectrometry imaging, Curr. Opin. Biotechnol, 43 (2017) 62–69. PubMed

Gemperline E, Rawson S, Li L, Optimization and Comparison of Multiple MALDI Matrix Application Methods for Small Molecule Mass Spectrometric Imaging, Analytical Chemistry, 86 (2014) 10030–10035. PubMed PMC

Vanbellingen QP, Castellanos A, Rodriguez-Silva M, Paudel I, Chambers JW, Fernandez-Lima FA, Analysis of Chemotherapeutic Drug Delivery at the Single Cell Level Using 3D-MSI-TOF-SIMS, Journal of The American Society for Mass Spectrometry, 27 (2016) 2033–2040. PubMed PMC

Henss A, Otto S-K, Schaepe K, Pauksch L, Lips KS, Rohnke M, High resolution imaging and 3D analysis of Ag nanoparticles in cells with ToF-SIMS and delayed extraction, Biointerphases, 13 (2018) 03B410. PubMed

Sämfors S, Ståhlman M, Klevstig M, Borén J, Fletcher JS, Localised lipid accumulation detected in infarcted mouse heart tissue using ToF-SIMS, Int. J. Mass Spectrom, DOI 10.1016/j.ijms.2017.09.012(2017). DOI

Tian H, Sparvero LJ, Amoscato AA, Bloom A, Bayır H, Kagan VE, Winograd N, Gas Cluster Ion Beam Time-of-Flight Secondary Ion Mass Spectrometry High-Resolution Imaging of Cardiolipin Speciation in the Brain: Identification of Molecular Losses after Traumatic Injury, Analytical Chemistry, 89 (2017) 4611–4619. PubMed PMC

Chandra S, Ausserer WA, Morrison GH, Evaluation of matrix effects in ion microscopic analysis of freeze-fractured, freeze-dried cultured cells, Journal of microscopy, 148 (1987) 223–229. PubMed

Slodzian G, Étude d’une méthode d’analyse locale chimique et isotopique utilisant l’émission ionique secondaire, Ann. Phys, 13 (1964) 591–648.

Brunelle A, Touboul D, Laprevote O, Biological tissue imaging with time-of-flight secondary ion mass spectrometry and cluster ion sources, Journal of Mass Spectrometry, 40 (2005) 985. PubMed

Touboul D, Halgand F, Brunelle A, Kersting R, Tallarek E, Hagenhoff B, Laprévote O, Tissue Molecular Ion Imaging by Gold Cluster Ion Bombardment, Analytical Chemistry, 76 (2004) 1550–1559. PubMed

Seah MP, Havelund R, Gilmore IS, Universal Equation for Argon Cluster Size-Dependence of Secondary Ion Spectra in SIMS of Organic Materials, The Journal of Physical Chemistry C, 118 (2014) 12862–12872.

Niehuis E, Möllers R, Rading D, Cramer HG, Kersting R, Analysis of organic multilayers and 3D structures using Ar cluster ions, Surface and Interface Analysis, 45 (2013) 158–162.

Seah MP, Universal Equation for Argon Gas Cluster Sputtering Yields, The Journal of Physical Chemistry C, 117 (2013) 12622–12632.

Fernandez-Lima FA, Post J, DeBord JD, Eller MJ, Verkhoturov SV, Della-Negra S, Woods AS, Schweikert EA, Analysis of Native Biological Surfaces Using a 100 kV Massive Gold Cluster Source, Analytical Chemistry, 83 (2011) 8448–8453. PubMed PMC

Adams KJ, DeBord JD, Fernandez-Lima F, Lipid specific molecular ion emission as a function of the primary ion characteristics in TOF-SIMS, Journal of Vacuum Science & Technology B, 34 (2016) 051804. PubMed PMC

Blain MG, Della-Negra S, Joret H, Le Beyec Y, Schweikert EA, Secondary-ion yields from surfaces bombarded with keV molecular and cluster ions, Physical Review Letters, 63 (1989) 1625. PubMed

Harris RD, Vanstipdonk NJ, Schweikert EA, keV Cluster Impacts: Prospects for Cluster-SIMS, Int. J. Mass Spectrom. Ion Proc., 174 (1998) 167–177.

Della-Negra S, Depauw J, Guillermier C, Schweikert EA, Massive clusters: Secondary emission from qkeV to qMeV. New emission processes? New SIMS probe?, Surface and Interface Analysis, 43 (2011) 62–65.

Fletcher JS, Lockyer NP, Vaidyanathan S, Vickerman JC, TOF-SIMS 3D Biomolecular Imaging of Xenopus laevis Oocytes Using Buckminsterfullerene (C60) Primary Ions, Analytical Chemistry, 79 (2007) 2199–2206. PubMed

Weibel D, Wong S, Lockyer N, Blenkinsopp P, Hill R, Vickerman JC, A C60 Primary Ion Beam System for Time of Flight Secondary Ion Mass Spectrometry:  Its Development and Secondary Ion Yield Characteristics, Analytical Chemistry, 75 (2003) 1754–1764. PubMed

van Stipdonk MJ, Harris RD, Schweikert EA, A Comparison of Desorption Yields from C+60 to Atomic and Polyatomic Projectiles at keV Energies, Rapid Communications in Mass Spectrometry, 10 (1996) 1987–1991.

Ninomiya S, Ichiki K, Yamada H, Nakata Y, Seki T, Aoki T, Matsuo J, Molecular depth profiling of multilayer structures of organic semiconductor materials by secondary ion mass spectrometry with large argon cluster ion beams, Rapid Communications in Mass Spectrometry, 23 (2009) 3264–3268. PubMed

Moritani K, Hashinokuchi M, Nakagawa J, Kashiwagi T, Toyoda N, Mochiji K, Extremely low-energy projectiles for SIMS using size-selected gas cluster ions, Appl. Surf. Sci, 255 (2008) 948–950.

Sheraz née Rabbani S, Barber A, Fletcher JS, Lockyer NP, Vickerman JC, Enhancing secondary ion yields in time of flight-secondary ion mass spectrometry using water cluster primary beams, Analytical chemistry, 85 (2013) 5654–5658. PubMed PMC

Tian H, Maciążek D, Postawa Z, Garrison BJ, Winograd N, CO2 Cluster Ion Beam, an Alternative Projectile for Secondary Ion Mass Spectrometry, Journal of the American Society for Mass Spectrometry, 27 (2016) 1476–1482. PubMed PMC

Bich C, Havelund R, Moellers R, Touboul D, Kollmer F, Niehuis E, Gilmore IS, Brunelle A, Argon Cluster Ion Source Evaluation on Lipid Standards and Rat Brain Tissue Samples, Anal. Chem. (Washington, DC, U. S.), 85 (2013) 7745. PubMed

Khalil SM, Roempp A, Pretzel J, Becker K, Spengler B, Phospholipid Topography of Whole-Body Sections of the Anopheles stephensi Mosquito, Characterized by High-Resolution Atmospheric-Pressure Scanning Microprobe Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Imaging, Anal. Chem. (Washington, DC, U. S.), 87 (2015) 11309–11316. PubMed

Phan Nhu TN, Fletcher John S, Sjövall P, Ewing Andrew G, ToF‐SIMS imaging of lipids and lipid related compounds in Drosophila brain, Surface and Interface Analysis, 46 (2014) 123–126. PubMed PMC

Bhandari D, Schott M, Römpp A, Vilcinskas A, Spengler B, Metabolite localization by atmospheric pressure high-resolution scanning microprobe matrix-assisted laser desorption/ionization mass spectrometry imaging in whole-body sections and individual organs of the rove beetle Paederus riparius, Analytical & Bioanalytical Chemistry, 407 (2015) 2189–2201. PubMed PMC

Kaftan F, Vrkoslav V, Kynast P, Kulkarni P, Böcker S, Cvačka J, Knaden M, Svatoš A, Mass spectrometry imaging of surface lipids on intact Drosophila melanogaster flies, Journal of Mass Spectrometry, 49 (2014) 223–232. PubMed

Zhou G, Pennington JE, Wells MA, Utilization of pre-existing energy stores of female Aedes aegypti mosquitoes during the first gonotrophic cycle, Insect Biochem. Mol. Biol, 34 (2004) 919–925. PubMed

Briegel H, Knusel I, Timmermann SE, Aedes aegypti: size, reserves, survival, and flight potential, J Vector Ecol, 26 (2001) 21–31. PubMed

Hagedorn HH, The Control of Vitellogenesis in the Mosquito, Aedes aegypti, American Zoologist, 14 (1974) 1207–1217.

Bhatt S, Gething PW, Brady OJ, Messina JP, Farlow AW, Moyes CL, Drake JM, Brownstein JS, Hoen AG, Sankoh O, Myers MF, George DB, Jaenisch T, Wint GRW, Simmons CP, Scott TW, Farrar JJ, Hay SI, The global distribution and burden of dengue, Nature, 496 (2013) 504. PubMed PMC

Weaver SC, Reisen WK, Present and future arboviral threats, Antiviral Research, 85 (2010) 328–345. PubMed PMC

Musso D, Gubler DJ, Zika Virus, Clinical Microbiology Reviews, 29 (2016) 487. PubMed PMC

Feachem RGA, Phillips AA, Hwang J, Cotter C, Wielgosz B, Greenwood BM, Sabot O, Rodriguez MH, Abeyasinghe RR, Ghebreyesus TA, Snow RW, Shrinking the malaria map: progress and prospects, The Lancet, 376 (2010) 1566–1578. PubMed PMC

Clements AN, The biology of mosquitoes Development, nutrition and reproduction, CABI Publishing, New York, NY, 1992.

Klowden MJ, Endocrine aspects of mosquito reproduction, Archives of Insect Biochemistry and Physiology, 35 (1997) 491–512.

Clifton ME, Noriega FG, Nutrient limitation results in juvenile hormone-mediated resorption of previtellogenic ovarian follicles in mosquitoes, Journal of Insect Physiology, 57 (2011) 1274–1281. PubMed PMC

Clifton ME, Noriega FG, The fate of follicles after a blood meal is dependent on previtellogenic nutrition and juvenile hormone in Aedes aegypti, Journal of Insect Physiology, 58 (2012) 1007–1019. PubMed PMC

Ziegler R, Ibrahim MM, Formation of lipid reserves in fat body and eggs of the yellow fever mosquito, Aedes aegypti, Journal of Insect Physiology, 47 (2001) 623–627. PubMed

Van Handel E, The obese mosquito, The Journal of Physiology, 181 (1965) 478–486. PubMed PMC

Chandra S, Morrison GH, Wolcott CC, Imaging intracellular elemental distribution and ion fluxes in cultured cells using ion microscopy: a freeze-fracture methodology, Journal of microscopy, 144 (1986) 15–37. PubMed

Berman ESF, Fortson SL, Checchi KD, Wu L, Felton JS, Wu KJJ, Kulp KS, Preparation of single cells for imaging/profiling mass spectrometry, Journal of the American Society for Mass Spectrometry, 19 (2008) 1230–1236. PubMed

Malm J, Giannaras D, Riehle MO, Gadegaard N, Sjövall P, Fixation and Drying Protocols for the Preparation of Cell Samples for Time-of-Flight Secondary Ion Mass Spectrometry Analysis, Analytical Chemistry, 81 (2009) 7197–7205. PubMed

Fletcher JS, Rabbani S, Henderson A, Lockyer NP, Vickerman JC, Three-dimensional mass spectral imaging of HeLa-M cells – sample preparation, data interpretation and visualisation, Rapid Communications in Mass Spectrometry, 25 (2011) 925–932. PubMed

Atella GC, Gondim KC, Machado EA, Medeiros MN, Silva-Neto MAC, Masuda H, Oogenesis and egg development in triatomines: a biochemical approach, An. Acad. Bras. Cienc, 77 (2005) 405–430. PubMed

Van Handel E, Fuel metabolism of the mosquito (Culex quinquefasciatus) embryo, Journal of Insect Physiology, 39 (1993) 831–833.

Briegel H, Hefti M, DiMarco E, Lipid metabolism during sequential gonotrophic cycles in large and small female Aedes aegypti, Journal of Insect Physiology, 48 (2002) 547–554. PubMed

DeBord JD, Smith DF, Anderton CR, Heeren RM, Pasa-Tolic L, Gomer RH, Fernandez-Lima FA, Secondary Ion Mass Spectrometry Imaging of Dictyostelium discoideum Aggregation Streams, Plos One, 9 (2014) e99319. PubMed PMC

Chandra S, 3D subcellular SIMS imaging in cryogenically prepared single cells, Appl. Surf. Sci, 231–232 (2004) 467–469.

Robinson MA, Graham DJ, Castner DG, ToF-SIMS Depth Profiling of Cells: z-Correction, 3D Imaging, and Sputter Rate of Individual NIH/3T3 Fibroblasts, Analytical Chemistry, 84 (2012) 4880–4885. PubMed PMC

Newman CF, Havelund R, Passarelli MK, Marshall PS, Francis I, West A, Alexander MR, Gilmore IS, Dollery CT, Intracellular Drug Uptake—A Comparison of Single Cell Measurements Using ToF-SIMS Imaging and Quantification from Cell Populations with LC/MS/MS, Analytical Chemistry, 89 (2017) 11944–11953. PubMed

Rao AN, Vandencasteele N, Gamble LJ, Grainger DW, High resolution epifluorescence and TOF-SIMS chemical imaging comparisons of single DNA microarray spots, Analytical chemistry, 84 (2012) 10628–10636. PubMed PMC

May CJ, Canavan HE, Castner DG, Quantitative X-ray Photoelectron Spectroscopy and Time-of-Flight Secondary Ion Mass Spectrometry Characterization of the Components in DNA, Analytical Chemistry, 76 (2004) 1114–1122. PubMed

Perez MH, Noriega FG, Aedes aegypti pharate 1st instar quiescence affects larval fitness and metal tolerance, Journal of insect physiology, 58 (2012) 824–829. PubMed PMC

Boggs CL, Ross CL, The Effect of Adult Food Limitation on Life History Traits in Speyeria Mormonia (Lepidoptera: Nymphalidae), Ecology, 74 (1993) 433–441.

Coupling Stable Isotope Labeling and Liquid Chromatography-Trapped Ion Mobility Spectrometry-Time-of-Flight-Tandem Mass Spectrometry for De Novo Mosquito Ovarian Lipid Studies

Following de novo triglyceride dynamics in ovaries of Aedes aegypti during the previtellogenic stage