Measuring the peptides in individual organelles with mass spectrometry

Rubakhin, Stanislav S.; Garden, Rebecca W.; Fuller, Robert R.; Sweedler, Jonathan V.

doi:10.1038/72622

Cited by 128 publications

(129 citation statements)

References 32 publications

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“…The Aplysia atrial gland is an exocrine organ producing as many as 20 different peptides that influence the reproductive behavior of the animal. The chemical composition of tissue homogenates, blots, and even single vesicles has been well characterized by MALDI MS [15]. This tissue was chosen for its ease of isolation, its well-characterized peptides, and more importantly, its reported homogenous nature that allowed sample preparation procedures to be easily assessed with a resolution of 100 m. The MALDI MS intensity image in Figure 4 of A-NTP (1397 m/z) obtained from a 30 m section of N 2 (l) frozen, SA electrosprayed atrial gland demonstrates several key points.…”

Section: Resultsmentioning

confidence: 99%

“…After the tissue was placed on the target, matrix was applied. The CHCA and SA matrices [15-20 mg/mL 3:2 (vol/vol) acetonitrile:0.3 % TFA] were deposited in multiple layers with electrospray using a modified homebuilt electrophoresis apparatus described previously [23] or by micro-spotting using a capillary micropipette device [15]. Briefly, for electrospray matrix deposition a syringe pump (Harvard Apparatus, South Natick, MA) delivered matrix to the tip of a 50 m i.d./150 m o.d.…”

Section: Sample Preparationmentioning

confidence: 99%

“…Even with the wealth of genomic information available, direct biochemical measures are often required to determine the presence and particular form of the neuropeptides present throughout the central nervous system (CNS). Direct MALDI MS analysis of molluscan and insect neurons [1][2][3][4][5][6][7][8][9][10][11][12][13], connective nerve tissues [14], and even single peptidergic vesicles [15] provides information about gene products and their processing to bioactive neuropeptides. Mass profiling individually isolated neurons is the most common method for obtaining spatial information such as the cell-specific distribution of peptides within brain regions known as ganglia.…”

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confidence: 99%

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Spatial profiling invertebrate ganglia using MALDI MS

Kruse

Sweedler

2003

J. Am. Soc. Mass Spectrom.

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108

100

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The ability of MALDI TOF MS to spatially map peptides and proteins directly from a tissue is an exciting advance to imaging mass spectrometry. Recent advances in instrumentation for MS have resulted in instruments capable of achieving several micron spatial resolution while acquiring high-resolution mass spectra. Currently, the ability to obtain high quality mass spectrometric images depends on sample preparation protocols that often result in limited spatial resolution. A number of sample preparation and matrix deposition protocols are evaluated for spatial profiling of Aplysia californica exocrine gland and neuronal tissues. Such samples are different from mammalian tissues, but make good targets for method optimization because of the wealth of biochemical information available on neuropeptide processing and distribution. Electrospray matrix deposition and a variety of freezing methods have been found to be optimum for these invertebrate tissues,

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Section: Resultsmentioning

confidence: 99%

Section: Sample Preparationmentioning

confidence: 99%

mentioning

confidence: 99%

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Spatial profiling invertebrate ganglia using MALDI MS

Kruse

Sweedler

2003

J. Am. Soc. Mass Spectrom.

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108

100

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“…For example, a single neuron from the snail Lymnaea stagnalis provided a sufficient quantity of material to identify all the peptides' products from both mRNA splice variants of the FMRFamide gene by MS (38). MALDI's compati- bility with heterogeneous solid samples allows the analysis of clusters of neurons; imaging of brain slices; and peptide profiling of smaller samples such as individual neurons, connectives, and even vesicles (33,39,40). How is spatial information on a particular peptide obtained?…”

Section: Neuropeptidesmentioning

confidence: 99%

Peer Reviewed: The Chemistry of Thought: Neurotransmitters in the Brain

Stuart¹,

Hummon²,

Sweedler³

2004

Anal. Chem.

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1 2 1 A I nside a neuron, clusters of molecules shuttle toward a cell membrane tightly packaged within vesicles. The vesicles fuse with the cell membrane, expelling the molecules into the extracellular space. As the molecules reach their targets, electrical currents fly, and the brain hums along.Without these chemical and electrical transformations, we would not remember the smell of coffee, recognize our child's face, or appreciate a delicate melody. The intricate signals and cascades induce thought, behavior, and memory. Analytical chemists are in on the act of understanding the compounds that drive human contemplation, including the reading of this sentence; what will be remembered after reading it; and even what emotions it will evoke. The goal of this article is to provide a general overview of the methods used to characterize neurotransmission in the brain, update recent specialized articles on brain chemistry, and complement articles about imaging MS and new electroanalytical methods that examine the role of dopamine in addiction (1-3).The brain is composed of nervous system cells, or neurons, which communicate with one another through ports called synapses. Cell-cell communication, or neurotransmission, is embodied through electrical impulses or chemical messengers. Although electrophysiological signaling is an important facet of brain function, we will focus on chemical signaling in the brain and the analytical methods used to study it.The Chemistry of Thought:

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“…Because the size of cells is quite limited, most MS approaches for single cells are based on MALDI-MS (23)(24)(25). MALDI-MS helped to discover hundreds of novel brain peptides, tens of which have been reported to be biologically active and related to animal behavior and functioning in models from arthropods and nematodes to mollusks (26).…”

mentioning

confidence: 99%

Single-neuron identification of chemical constituents, physiological changes, and metabolism using mass spectrometry

Zhu

Zou

Wang

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

105

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The use of single-cell assays has emerged as a cutting-edge technique during the past decade. Although single-cell mass spectrometry (MS) has recently achieved remarkable results, deep biological insights have not yet been obtained, probably because of various technical issues, including the unavoidable use of matrices, the inability to maintain cell viability, low throughput because of sample pretreatment, and the lack of recordings of cell physiological activities from the same cell. In this study, we describe a patch clamp/MS-based platform that enables the sensitive, rapid, and in situ chemical profiling of single living neurons. This approach integrates modified patch clamp technique and modified MS measurements to directly collect and detect nanoliter-scale samples from the cytoplasm of single neurons in mice brain slices. Abundant possible cytoplasmic constituents were detected in a single neuron at a relatively fast rate, and over 50 metabolites were identified in this study. The advantages of direct, rapid, and in situ sampling and analysis enabled us to measure the biological activities of the cytoplasmic constituents in a single neuron, including comparing neuron types by cytoplasmic chemical constituents; observing changes in constituent concentrations as the physiological conditions, such as age, vary; and identifying the metabolic pathways of small molecules.single neuron | mass spectrometry | metabolism | patch clamp

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Measuring the peptides in individual organelles with mass spectrometry

Cited by 128 publications

References 32 publications

Spatial profiling invertebrate ganglia using MALDI MS

Spatial profiling invertebrate ganglia using MALDI MS

Peer Reviewed: The Chemistry of Thought: Neurotransmitters in the Brain

Single-neuron identification of chemical constituents, physiological changes, and metabolism using mass spectrometry

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