Tissue Localization of Glycosphingolipid Accumulation in a Gaucher Disease Mouse Brain by LC-ESI-MS/MS and High-Resolution MALDI Imaging Mass Spectrometry

To better understand regional brain glycosphingolipid (GSL) accumulation in Gaucher disease (GD) and its relationship to neuropathology, a feasibility study using mass spectrometry and immunohistochemistry was conducted using brains derived from a GD mouse model (4L/PS/NA) homozygous for a mutant GCase (V394L [4L]) and expressing a prosaposin hypomorphic (PS-NA) transgene. Whole brains from GD and control animals were collected using one hemisphere for MALDI FTICR IMS analysis and the other for quantitation by LC-ESI-MS/MS. MALDI IMS detected several HexCers across the brains. Comparison with the brain hematoxylin and eosin (H&E) revealed differential signal distributions in the midbrain, brain stem, and CB of the GD brain versus the control. Quantitation of serial brain sections with LC-ESI-MS/MS supported the imaging results, finding the overall HexCer levels in the 4L/PS-NA brains to be four times higher than the control. LC-ESI-MS/MS also confirmed that the elevated hexosyl isomers were glucosylceramides rather than galactosylceramides. MALDI imaging also detected differential analyte distributions of lactosylceramide species and gangliosides in the 4L/PS-NA brain, which was validated by LC-ESI-MS/MS. Immunohistochemistry revealed regional inflammation, altered autophagy, and defective protein degradation correlating with regions of GSL accumulation, suggesting that specific GSLs may have distinct neuropathological effects.

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Defining Changes in the Spatial Distribution and Composition of Brain Lipids in the Shiverer and Cuprizone Mouse Models of Myelin Disease

Journal of Histochemistry & Cytochemistry ◽

10.1369/0022155418815860 ◽

2018 ◽

Vol 67 (3) ◽

pp. 203-219 ◽

Cited By ~ 2

Author(s):

Rajanikanth J. Maganti ◽

Xiaoping L. Hronowski ◽

Robert W. Dunstan ◽

Brian T. Wipke ◽

Xueli Zhang ◽

...

Keyword(s):

Mass Spectrometry ◽

Spatial Distribution ◽

Mouse Model ◽

Morphological Changes ◽

Imaging Mass Spectrometry ◽

Ionization Mass ◽

Brain Lipids ◽

Esi Ms ◽

Maldi Ims ◽

Lipid Species

Myelin is composed primarily of lipids and diseases affecting myelin are associated with alterations in its lipid composition. However, correlation of the spatial (in situ) distribution of lipids with the disease-associated compositional and morphological changes is not well defined. Herein we applied high resolution matrix-assisted laser desorption ionization imaging mass spectrometry (MALDI-IMS), immunohistochemistry (IHC), and liquid chromatography–electrospray ionization–mass spectrometry (LC-ESI-MS) to evaluate brain lipid alterations in the dysmyelinating shiverer (Shi) mouse and cuprizone (Cz) mouse model of reversible demyelination. MALDI-IMS revealed a decrease in the spatial distribution of sulfatide (SHexCer) species, SHexCer (d42:2), and a phosphatidylcholine (PC) species, PC (36:1), in white matter regions like corpus callosum (CC) both in the Shi mouse and Cz mouse model. Changes in these lipid species were restored albeit not entirely upon spontaneous remyelination after demyelination in the Cz mouse model. Lipid distribution changes correlated with the local morphological changes as confirmed by IHC. LC-ESI-MS analyses of CC extracts confirmed the MALDI-IMS derived reductions in SHexCer and PC species. These findings highlight the role of SHexCer and PC in preserving the normal myelin architecture and our experimental approaches provide a morphological basis to define lipid abnormalities relevant to myelin diseases.

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MALDI Imaging Mass Spectrometry (MALDI-IMS)―Application of Spatial Proteomics for Ovarian Cancer Classification and Diagnosis

International Journal of Molecular Sciences ◽

10.3390/ijms12010773 ◽

2011 ◽

Vol 12 (1) ◽

pp. 773-794 ◽

Cited By ~ 69

Author(s):

Johan O. R. Gustafsson ◽

Martin K. Oehler ◽

Andrew Ruszkiewicz ◽

Shaun R. McColl ◽

Peter Hoffmann

Keyword(s):

Mass Spectrometry ◽

Ovarian Cancer ◽

Imaging Mass Spectrometry ◽

Cancer Classification ◽

Maldi Imaging ◽

Maldi Ims ◽

Maldi Imaging Mass Spectrometry

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Dual polarity MALDI imaging mass spectrometry on the same pixel points reveals spatial lipid localizations at high-spatial resolutions in rat small intestine

Analytical Methods ◽

10.1039/c8ay00645h ◽

2018 ◽

Vol 10 (21) ◽

pp. 2428-2435 ◽

Cited By ~ 9

Author(s):

Ibrahim Kaya ◽

Eva Jennische ◽

Stefan Lange ◽

Per Malmberg

Keyword(s):

Mass Spectrometry ◽

Imaging Mass Spectrometry ◽

Laser Desorption ◽

Maldi Imaging ◽

Jejunal Mucosa ◽

Rat Small Intestine ◽

Laser Desorption Ionization ◽

Desorption Ionization ◽

Maldi Ims ◽

Dual Polarity

Sensitive laser desorption/ionization obtained via a sublimation-coated 1,5 diaminonaphthalene (1,5-DAN) matrix allowed dual polarity MALDI-IMS analysis on the same pixel points across the jejunal mucosa.

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Optimized Workflow for High Spatial Resolution MALDI Imaging Mass Spectrometry of Fresh-Frozen Bone

10.1101/2021.10.01.462831 ◽

2021 ◽

Author(s):

Christopher J. Good ◽

Elizabeth K. Neumann ◽

Casey E. Butrico ◽

James E. Cassat ◽

Richard M. Caprioli ◽

...

Keyword(s):

Mass Spectrometry ◽

Bone Marrow ◽

Adipose Tissue ◽

Spatial Resolution ◽

High Spatial Resolution ◽

Imaging Mass Spectrometry ◽

Maldi Imaging ◽

Maldi Ims ◽

Fresh Frozen ◽

Frozen Bone

Bone and bone marrow are vital to mammalian structure, movement, and immunity. These tissues are also commonly subjected to pathological alterations giving rise to debilitating diseases like rheumatoid arthritis, osteoporosis, osteomyelitis, and cancer. Technologies such as matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry (IMS) enable the discovery of spatially resolved chemical information in biological tissue samples to help elucidate the complex molecular processes underlying pathology. Traditionally, preparation of native osseous tissue for MALDI IMS has been difficult due to the mineralized composition and heterogenous morphology of the tissue, and compensation for these challenges with decalcification and fixation protocols can remove or delocalize molecular species. Here, sample preparation methods were advanced to enable multimodal MALDI IMS of undecalcified, fresh-frozen murine femurs allowing the distribution of endogenous lipids to be linked to specific tissue structures and cell types. Adhesive-bound bone sections were mounted onto ITO coated glass slides with a microscopy-compatible glue and freeze-dried to minimize artificial bone marrow damage. Subliming matrix does not induce further bone marrow cracks, and recrystallizing the deposited matrix improves lipid signal. High spatial resolution (10 μm) MALDI IMS was leveraged to characterize lipid distributions in fresh-frozen bone, and complementary microscopy modalities aided tissue and cell assignments. For example, various phosphatidylcholines localize to bone marrow, adipose tissue, marrow adipose tissue, and muscle. Furthermore, we discovered that [sphingomyelin(42:1) + H]+ was abundant in megakaryocytes, whereas [sphingomyelin(42:2) + H]+ was diminished in this cell type. These data reflect the vast molecular and cellular heterogeneity indicative of the bone marrow and the soft tissue surrounding the femur. Therefore, this application of multimodal MALDI IMS has the potential to advance bone-related biomedical research by offering deep molecular coverage in a preserved native bone microenvironment.

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