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by claude@2026-07, 2026-07-03
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The paper studied astrocyte functional heterogeneity by using Microscoop Mint, a microscopy-guided spatial proteomics platform that profiles protein expression from subcellular, region-specific samples in paraformaldehyde-fixed, OCT-embedded mouse brain tissue via LC-MS/MS. In the cerebral cortex and hippocampus, it identified distinct regional-associated astrocyte proteomic signatures and proposed novel candidate protein markers, including preferential expression of MINK1 in hippocampal astrocytes and PLEKHB1 in cortical astrocytes, for later immunofluorescence validation. The main caveat is that the work is focused on protein profiling in fixed mouse brain tissue, with subsequent confirmation stated as a validation step rather than demonstrated for the candidate markers. The paper does not explicitly discuss endometriosis or adenomyosis; it was included in the corpus via a keyword match in the upstream search index.
Abstract
ABSTRACT Astrocytes are a subset of glial cells in the central nervous system (CNS) that support numerous processes essential for brain function. Their functional diversity is thought to arise from specialized subpopulations with distinct molecular profiles. Although single-cell and single-nucleus RNA sequencing (scRNA-seq and snRNA-seq) have greatly advanced our understanding of astrocyte transcriptomic heterogeneity, mRNA abundance does not always correlate with protein levels because of post-transcriptional and translational regulation. Therefore, studying protein profiles remains essential to accurately capture astrocyte functional states and heterogeneity. Here, we used Microscoop Mint, a microscopy-guided spatial proteomics platform that integrates subcellular, region-specific sample preparation with LC-MS/MS-based mass spectrometry, enabling direct protein profiling of astrocytes in paraformaldehyde-fixed, optimal cutting temperature (OCT)-embedded mouse brain tissue. By applying this approach, we uncovered distinct regional-associated astrocyte proteomic signatures in the cerebral cortex and hippocampus and selected novel candidate protein markers for subsequent validation by immunofluorescence. Notably, MINK1 and PLEKHB1 showed preferential expression in hippocampal and cortical astrocytes, respectively, highlighting their potential as region-specific astrocyte markers. Overall, this strategy enables high-precision, unbiased spatial proteomic discovery at subcellular resolution, providing a powerful framework for linking molecular diversity to functional specialization in astrocyte biology.
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ABSTRACT
Astrocytes are a subset of glial cells in the central nervous system (CNS) that support numerous processes essential for brain function. Their functional diversity is thought to arise from specialized subpopulations with distinct molecular profiles. Although single-cell and single-nucleus RNA sequencing (scRNA-seq and snRNA-seq) have greatly advanced our understanding of astrocyte transcriptomic heterogeneity, mRNA abundance does not always correlate with protein levels because of post-transcriptional and translational regulation. Therefore, studying protein profiles remains essential to accurately capture astrocyte functional states and heterogeneity. Here, we used Microscoop Mint, a microscopy-guided spatial proteomics platform that integrates subcellular, region-specific sample preparation with LC-MS/MS-based mass spectrometry, enabling direct protein profiling of astrocytes in paraformaldehyde-fixed, optimal cutting temperature (OCT)-embedded mouse brain tissue. By applying this approach, we uncovered distinct regional-associated astrocyte proteomic signatures in the cerebral cortex and hippocampus and selected novel candidate protein markers for subsequent validation by immunofluorescence. Notably, MINK1 and PLEKHB1 showed preferential expression in hippocampal and cortical astrocytes, respectively, highlighting their potential as region-specific astrocyte markers. Overall, this strategy enables high-precision, unbiased spatial proteomic discovery at subcellular resolution, providing a powerful framework for linking molecular diversity to functional specialization in astrocyte biology.
Competing Interest Statement
Patent applications related to the subject matter of this publication have been filed. All authors declare that they are current employees of Syncell Inc.
ABBREVIATIONS
- ROI
- region of interest
- FOV
- field of view
- GFAP
- glial fibrillary acidic protein
- CNS
- central nervous system
- DIA
- data-independent acquisition
- PBS
- phosphate buffered saline
- PBST
- phosphate buffered saline containing 0.1% Triton X-100
- DAPI
- 4′,6-diamidino-2-phenylindole.
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