In situ molecular imaging of proteins in tissues using mass spectrometry

Hardesty, William M.; Caprioli, Richard M.

doi:10.1007/s00216-008-1972-5

Cited by 37 publications

(29 citation statements)

References 12 publications

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“…Mass spectral analysis of a trypsin digest of purified recombinant CYP2C9 identified diagnostic peptides at m/z 1491 (ANRGFGIVFSNGKK), 2415 (VQEEIERVIGR), and 3017 (YALLLLLKHPEVTAKVQ EEIERVIGR). Scanning was done at these diagnostic m/z values, and two-dimensional images were acquired, normalized, and visualized with FlexImaging (38).…”

Section: Methodsmentioning

confidence: 99%

“…For mass spectral imaging, serial sections of the renal carcinoma used for immunofluorescence (12 m each) were thawmounted onto gold-coated MALDI plates, washed with ethanol (38), and stored ϳ2 h under vacuum. A robotic spotter (Portrait, LabCyte) was used to deposit in fifty cycles a solution of porcine trypsin (0.8 mg/ml) in 16 mM HOAc, 84 mM NH 4 HCO 3 (one drop per cycle, fly-by mode) across the tissue section at a spot center-to-center spacing of 300 m, followed by 30 cycles of 40 mg/ml 2,5-dihydroxybenzoic acid matrix in 50:50:0.1% (v/v) acetonitrile:water:trifluoroacetic acid.…”

Section: Methodsmentioning

confidence: 99%

“…However, although the relative contributions of these isoforms to the biosynthesis of pro-angiogenic EETs remain undetermined, the studies summarized above point to CYP2C9 as a pro-angiogenic epoxygenase and a potential target for the anti-tumorigenic effects of ligand activated human PPAR␣. Therefore, to further document the relevance of our mouse studies to human cancer, we utilized mass spectrometry imaging techniques (38) to identify and map the expression and distribution of CYP2C9 in a human clear cell renal carcinoma. This type of carcinoma was selected for these studies, because its frozen sections retain most of their morphological details after in situ proteolysis.…”

Section: Activation Of the Human Ppar␣ In Ppar␣-humanized Mice Inhibimentioning

confidence: 99%

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The Anti-tumorigenic Properties of Peroxisomal Proliferator-activated Receptor α Are Arachidonic Acid Epoxygenase-mediated

Pozzi

Popescu

Yang

et al. 2010

Journal of Biological Chemistry

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104

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Despite advances in diagnosis and treatment, cancer continues to pose major clinical challenges and is the focus of efforts to develop therapies that combine effectiveness with low toxicity. Among these, angiogenesis is a current target of attempts aimed to reduce tumor vascularization and growth (1, 2), because it holds a promise for more effective and better tolerated approaches for cancer treatment. The peroxisomal proliferator-activated nuclear receptors (PPARs), 2 i.e. PPAR␣, PPAR␥, and PPAR␦, control the transcription of genes mostly involved in the regulation of lipid metabolism and energy homeostasis. Although several roles for the PPAR␦ and -␥ isotypes in the pathophysiology of cancer have been proposed (3-5), recent animal studies have identified PPAR␣ ligands as effective inhibitors of tumor angiogenesis and growth (6 -8).The potential significance of these findings has been emphasized by epidemiological data (9 -12), and by studies in human cancer cell lines (13-16), suggesting that PPAR␣ ligands such as Fenofibrate and Bezafibrate may have beneficial effects in the prognosis of human cancer. A unique feature of these PPAR␣ ligands is that, through their long history of clinical use as hypolipidemic drugs, they have been shown to be well tolerated and to have limited side effects and/or toxicity, indicating that they could serve as targets for the development of novel, safer, and with low toxicity anti-cancer treatments.The PPAR␣-mediated transcriptional regulation of members of the CYP2C gene subfamily of cytochrome P450s is well established (17, 18), as is the role of these enzymes in the metabolism and bio-activation of arachidonic acid (AA) (19). The CYP2C epoxygenases metabolize AA to 5,8,11,and 14,, and these metabolites have been characterized as pro-angiogenic lipids in vitro (20 -22) and in vivo (23). The demonstration that the anti-tumorigenic effects of PPAR␣ ligand activation were associated with reductions in the endothelial expression of the murine Cyp2c44 epoxygenase and in the levels of plasma and endothelial EETs (6), suggested pro-angiogenic and pro-tumorigenic roles for this epoxygenase, and pointed to this enzyme as a target of the anti-tumorigenic effects resulting from PPAR␣ activation (6). The murine Cyp2c44 epoxygenase generates 11,12-and 14,15-EET as its major products (24), is expressed in endothelial cells (6), and is under PPAR␣ transcriptional control (6). Similarly, human CYP2C8 and CYP2C9, catalytic homologues of murine Cyp2c44 (25), have been identified as endothelial epoxygenases (21,22), and their participation in

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Activation Of the Human Ppar␣ In Ppar␣-humanized Mice Inhibimentioning

confidence: 99%

See 1 more Smart Citation

The Anti-tumorigenic Properties of Peroxisomal Proliferator-activated Receptor α Are Arachidonic Acid Epoxygenase-mediated

Pozzi

Popescu

Yang

et al. 2010

Journal of Biological Chemistry

Self Cite

104

View full text Add to dashboard Cite

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“…Many of the advances in MALDI-MSI, developed by Caprioli and coworkers (5,6), have been applied to localization of peptides and proteins within tissues. This technique has been shown to generate abundant secondary ions from proteins in tissues, though only after lipids, which in many cases generate the most abundant ion current, were removed from the tissue.…”

mentioning

confidence: 99%

Imaging of lipid species by MALDI mass spectrometry

Murphy

Hankin

Barkley

2009

Journal of Lipid Research

267

241

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Recent developments in MALDI have enabled direct detection of lipids as intact molecular species present within cellular membranes. Abundant lipid-related ions are produced from the direct analysis of thin tissue slices when sequential spectra are acquired across a tissue surface that has been coated with a MALDI matrix. The lipid-derived ions can often be distinguished from other biomolecules because of the significant mass defect that these ions present due to the large number of covalently bound hydrogen atoms in hydrophobic molecules such as lipids. Collisional activation of the molecular ions can be used to determine the lipid family and often structurally define the molecular species. Specific examples in the detection of phospholipids, sphingolipids, and glycerolipids are presented with images of mouse brain and kidney tissue slices. Regional distribution of many different lipid molecular species and Na

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“…maging mass spectrometry is a widely used tool for detecting the molecular composition of sample surfaces [1][2][3][4][5][6] with a 2D resolution in the μm range [7]. The technology has been proven valuable for many fields of application, ranging from diagnostics in microchip production to analysis of tissue samples.…”

mentioning

confidence: 99%

Digital Imaging Mass Spectrometry

Bamberger

Renz

Bamberger

2011

J. Am. Soc. Mass Spectrom.

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Methods to visualize the two-dimensional (2D) distribution of molecules by mass spectrometric imaging evolve rapidly and yield novel applications in biology, medicine, and material surface sciences. Most mass spectrometric imagers acquire high mass resolution spectra spot-by-spot and thereby scan the object's surface. Thus, imaging is slow and image reconstruction remains cumbersome. Here we describe an imaging mass spectrometer that exploits the true imaging capabilities by ion optical means for the time of flight mass separation. The mass spectrometer is equipped with the ASIC Timepix chip as an array detector to acquire the position, mass, and intensity of ions that are imaged by matrix-assisted laser desorption/ionization (MALDI) directly from the target sample onto the detector. This imaging mass spectrometer has a spatial resolving power at the specimen of (84±35)μm with a mass resolution of 45 and locates atoms or organic compounds on a surface area up to~2 cm 2 . Extended laser spots of~5 mm 2 on structured specimens allows parallel imaging of selected masses. The digital imaging mass spectrometer proves high hit-multiplicity, straightforward image reconstruction, and potential for high-speed readout at 4 kHz or more. This device demonstrates a simple way of true image acquisition like a digital photographic camera. The technology may enable a fast analysis of biomolecular samples in near future.

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In situ molecular imaging of proteins in tissues using mass spectrometry

Cited by 37 publications

References 12 publications

The Anti-tumorigenic Properties of Peroxisomal Proliferator-activated Receptor α Are Arachidonic Acid Epoxygenase-mediated

The Anti-tumorigenic Properties of Peroxisomal Proliferator-activated Receptor α Are Arachidonic Acid Epoxygenase-mediated

Imaging of lipid species by MALDI mass spectrometry

Digital Imaging Mass Spectrometry

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