Regional N-glycan and lipid analysis from tissues using MALDI-mass spectrometry imaging

Stanback, Alexandra E.; Conroy, Lindsey R.; Young, Lyndsay E.A.; Hawkinson, Tara R.; Markussen, Kia H.; Clarke, Harrison A.; Allison, Derek B.; Sun, Ramon C.

doi:10.1016/j.xpro.2021.100304

Cited by 23 publications

(29 citation statements)

References 6 publications

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“…The most common matrices for the MSI analysis of glycome are 2,5-dihydroxybenzoic acid (DHB) and α-cyano-4-hydroxycinnamic acid (CHCA). Usually, the matrices are sprayed on the tissue using an automated robotic sprayer such as HTX M5 Sprayer TM ( Stanback et al, 2021 ), HTX TM-Sprayer TM ( Shi et al, 2019 ), ImagePrep sprayer ( Powers et al, 2014 ). After matrix coating, the tissue sections are sent to an MSI instruments for data acquisition and analysis.…”

Section: Methodology For Glycome Msi In the Brainmentioning

confidence: 99%

“…They also reported that enormously high structural information of complex N-glycans was obtained directly from thin tissue sections in the human cerebellum and upon low-energy collision-induced dissociation tandem MS. However, MALDI-MSI techniques are capable of visualizing hundreds of glycome with spatial information in a single brain tissue followed by PNGase F treated or non-treated sample ( Figure 3A ; Stanback et al, 2021 ), which is limited by the typical antibody staining methods. Figure 3A showed the representative ion images of Figure 3A-iv Hex5HexNAc4Fuc1 at m/z 1809, ( Figure 3A-v ) Hex7HexNAc2 at m/z 1581, ( Figure 3A-vi ) Hex9HexNAc2 at m/z 1905, and ( Figure 3A-vii ) the overlay ion image of those N-glycans in FFPE and FF mouse brain tissues by the standard MALDI-MSI protocols described by Stanback et al, 2021 .…”

Section: Msi Analysis For Glycome In the Brainmentioning

confidence: 99%

“…Scale bar, 2 mm (upper panel), 500 μm (lower panel). The figures of panels (A,B) are reprinted and modified from Stanback et al (2021) and Andres et al (2020) , respectively, with free-reuse permission.…”

Section: Msi Analysis For Glycome In the Brainmentioning

confidence: 99%

“…They also reported that enormously high structural information of complex N-glycans was obtained directly from thin tissue sections in the human cerebellum and upon lowenergy collision-induced dissociation tandem MS. However, MALDI-MSI techniques are capable of visualizing hundreds of glycome with spatial information in a single brain tissue followed by PNGase F treated or non-treated sample (Figure 3A; Stanback et al, 2021), which is limited by the typical antibody staining methods.…”

Section: Msi Analysis For Glycome In the Brainmentioning

confidence: 99%

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Mass Spectrometry Imaging for Glycome in the Brain

et al. 2021

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Glycans are diverse structured biomolecules that play crucial roles in various biological processes. Glycosylation, an enzymatic system through which various glycans are bound to proteins and lipids, is the most common and functionally crucial post-translational modification process. It is known to be associated with brain development, signal transduction, molecular trafficking, neurodegenerative disorders, psychopathologies, and brain cancers. Glycans in glycoproteins and glycolipids expressed in brain cells are involved in neuronal development, biological processes, and central nervous system maintenance. The composition and expression of glycans are known to change during those physiological processes. Therefore, imaging of glycans and the glycoconjugates in the brain regions has become a “hot” topic nowadays. Imaging techniques using lectins, antibodies, and chemical reporters are traditionally used for glycan detection. However, those techniques offer limited glycome detection. Mass spectrometry imaging (MSI) is an evolving field that combines mass spectrometry with histology allowing spatial and label-free visualization of molecules in the brain. In the last decades, several studies have employed MSI for glycome imaging in brain tissues. The current state of MSI uses on-tissue enzymatic digestion or chemical reaction to facilitate successful glycome imaging. Here, we reviewed the available literature that applied MSI techniques for glycome visualization and characterization in the brain. We also described the general methodologies for glycome MSI and discussed its potential use in the three-dimensional MSI in the brain.

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Section: Methodology For Glycome Msi In the Brainmentioning

confidence: 99%

Section: Msi Analysis For Glycome In the Brainmentioning

confidence: 99%

Section: Msi Analysis For Glycome In the Brainmentioning

confidence: 99%

Section: Msi Analysis For Glycome In the Brainmentioning

confidence: 99%

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Mass Spectrometry Imaging for Glycome in the Brain

et al. 2021

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“…Formalin-fixed paraffin embedded lungs (FFPE) were processed as previously described 16 . In brief, FFPE lungs were sectioned at 5 μ m on glass slides.…”

Section: Methodsmentioning

confidence: 99%

Cervical Spinal Cord Injury Leads to Injury and Metabolic Dysfunction of the Lung

Huffman

Dong

Clarke

et al. 2021

Preprint

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High-cervical spinal cord injury (SCI) often disrupts respiratory motor pathways and disables breathing in the affected population. Moreover, cervically injured individuals are at risk for developing acute lung injury (ALI), which predicts substantial mortality rates. While the correlation between ALI and SCI has been found in the clinical setting, the field lacks an animal model to interrogate the fundamental biology of this relationship. To begin to address this gap, we performed an experimental cervical SCI and assessed lung injury in adult rats. We demonstrate that animals display signs of ALI two weeks post-SCI. We also observed aberrant N-glycan metabolism determined by matrix-assisted laser desorption/ionization mass spectrometry imaging. Collectively, we establish for the first time a model of ALI after SCI at an acute time point that can be used to monitor the progression of lung damage, as well as identify potential targets to ameliorate ALI.

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In situ spatial glycomic imaging of mouse and human Alzheimer's disease brains

Hawkinson

Clarke

Young

et al. 2021

Alzheimer's & Dementia

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N‐linked protein glycosylation in the brain is an understudied facet of glucose utilization that impacts a myriad of cellular processes including resting membrane potential, axon firing, and synaptic vesicle trafficking. Currently, a spatial map of N‐linked glycans within the normal and Alzheimer's disease (AD) human brain does not exist. A comprehensive analysis of the spatial N‐linked glycome would improve our understanding of brain energy metabolism, linking metabolism to signaling events perturbed during AD progression, and could illuminate new therapeutic strategies. Herein we report an optimized in situ workflow for enzyme‐assisted, matrix‐assisted laser desorption and ionization (MALDI) mass spectrometry imaging (MSI) of brain N‐linked glycans. Using this workflow, we spatially interrogated N‐linked glycan heterogeneity in both mouse and human AD brains and their respective age‐matched controls. We identified robust regional‐specific N‐linked glycan changes associated with AD in mice and humans. These data suggest that N‐linked glycan dysregulation could be an underpinning of AD pathologies.

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Regional N-glycan and lipid analysis from tissues using MALDI-mass spectrometry imaging

Cited by 23 publications

References 6 publications

Mass Spectrometry Imaging for Glycome in the Brain

Mass Spectrometry Imaging for Glycome in the Brain

Cervical Spinal Cord Injury Leads to Injury and Metabolic Dysfunction of the Lung

In situ spatial glycomic imaging of mouse and human Alzheimer's disease brains

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