The restricted N-glycome of neurons is programmed during differentiation DOI Creative Commons
Katherine Kiwimagi, Maxence Noël, Murat Çetinbaş

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Oct. 17, 2024

Abstract The protein glycome of individual cell types in the brain is unexplored, despite critical function these modifications development and disease. In aggregate, most abundant asparagine (N-) linked glycans adult are high mannose structures, specifically Man 5 GlcNAc 2 (Man-5), which normally exits ER for further processing Golgi. Mannose structures uncommon other organs often overlooked or excluded studies. To understand cell-specific contributions to unique N-glycome its abundance Man-5, we performed RNAseq MALDI-MS TOF N-glycomics at several timepoints during differentiation multiple types. this end, homogeneous cultures glutamatergic neurons, GABAergic brain-specific endothelial cells were generated from monoclonal human inducible pluripotent stem (hiPSCs) through cellular reprogramming. Small molecule induction stably integrated synthetic transcription units driving morphogen expression differentiated with distinct patterns mirroring intact tissue. Comparing uninduced hiPSCs each type revealed identical transcriptomic glycomic profiles before differentiation, low quantities Man-5. N-glycans became Man-5 immediate precursor Man-6, presence transcripts encoding enzymes their subsequent modification. Differentiation showed an opposite effect, displaying complex terminal late secretory pathway. These results confirm that restricted profile programmed into neuronal regulation independent transcriptome under tight evolutionary constraint.

Language: Английский

O-GalNAc glycans are enriched in neuronal tracts and regulate nodes of Ranvier DOI Creative Commons
Maxence Noël, Suttipong Suttapitugsakul, Richard D. Cummings

et al.

Proceedings of the National Academy of Sciences, Journal Year: 2025, Volume and Issue: 122(9)

Published: Feb. 25, 2025

Protein O-glycosylation is a critical modification in the brain, as genetic variants pathway are associated with common and severe neuropsychiatric phenotypes. However, little known about most abundant O-glycans mammalian which N-acetylgalactosamine (O-GalNAc) linked. Here, we determined spatial localization, protein carriers, cellular function of O-GalNAc glycans mouse brain. We observed striking enrichment neuronal tracts, specifically at nodes Ranvier, specialized structures involved signal propagation Glycoproteomic analysis revealed that more than half identified were present on chondroitin sulfate proteoglycans termed lecticans, display both domain regional heterogeneity. Inhibition synthesis neurons reduced binding Siglec-4, regulator neurite growth, shortened length Ranvier. This work establishes brain will inform future studies their role development disease.

Language: Английский

Citations

0
O-GalNAc glycans enrich in white matter tracts and regulate nodes of Ranvier DOI Creative Commons
Maxence Noël, Suttipong Suttapitugsakul, Richard D. Cummings

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: May 9, 2024

Abstract Protein O-glycosylation is a critical modification in the brain, as genetic variants pathway are associated with both common and severe neuropsychiatric phenotypes. However, little known about most abundant type of O-glycan mammalian which O-GalNAc linked. Here, we determined spatial localization, protein carriers, cellular function glycans mouse brain. We observed striking enrichment white matter tracts at nodes Ranvier. Glycoproteomic analysis revealed that more than half identified were present on chondroitin sulfate proteoglycans termed lecticans, display domain site-specific heterogeneity. Though ablation single cell failed to replicate phenotypes seen congenital disorders, inhibition synthesis neurons reduced binding Siglec-4, regulator neurite growth, shortened length This work highlights new brain will inform future studies their role development disease.

Language: Английский

Citations

0
Unique Glycans in Synaptic Glycoproteins in Mouse Brain DOI
Maxence Noël, Suttipong Suttapitugsakul, Mohui Wei

et al.

ACS Chemical Neuroscience, Journal Year: 2024, Volume and Issue: 15(21), P. 4033 - 4045

Published: Oct. 14, 2024

The synapse is an essential connection between neuronal cells in which the membrane and secreted glycoproteins regulate neurotransmission. post-translational modifications of with carbohydrates, although for their functions as well specific localization, are not understood. Oddly, whereas galactose addition to required functions, galactosylation severely restricted Asn-linked on N-glycans brain, genetic evidence highlights important roles brain development. To explore this novel glycosylation, we exploited orthogonal technology a biotinylated sialic acid derivative (CMP-biotin-Sia) transferred terminally galactosylated proteins by recombinant sialyltransferase (rST6Gal1). This approach allowed us identify carrier localization sections. Immunohistochemical analysis sections demonstrates that they largely positioned pre- postsynaptic membranes. Consistent positioning, glycoproteomic analyses labeled identified number them involved synaptic function, cell adhesion, extracellular matrix interactions. discovery these N-glycoproteins relative confinement synapses provide insights into unusual nature protein glycosylation brain.

Language: Английский

Citations

0
The restricted N-glycome of neurons is programmed during differentiation DOI Creative Commons
Katherine Kiwimagi, Maxence Noël, Murat Çetinbaş

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Oct. 17, 2024

Abstract The protein glycome of individual cell types in the brain is unexplored, despite critical function these modifications development and disease. In aggregate, most abundant asparagine (N-) linked glycans adult are high mannose structures, specifically Man 5 GlcNAc 2 (Man-5), which normally exits ER for further processing Golgi. Mannose structures uncommon other organs often overlooked or excluded studies. To understand cell-specific contributions to unique N-glycome its abundance Man-5, we performed RNAseq MALDI-MS TOF N-glycomics at several timepoints during differentiation multiple types. this end, homogeneous cultures glutamatergic neurons, GABAergic brain-specific endothelial cells were generated from monoclonal human inducible pluripotent stem (hiPSCs) through cellular reprogramming. Small molecule induction stably integrated synthetic transcription units driving morphogen expression differentiated with distinct patterns mirroring intact tissue. Comparing uninduced hiPSCs each type revealed identical transcriptomic glycomic profiles before differentiation, low quantities Man-5. N-glycans became Man-5 immediate precursor Man-6, presence transcripts encoding enzymes their subsequent modification. Differentiation showed an opposite effect, displaying complex terminal late secretory pathway. These results confirm that restricted profile programmed into neuronal regulation independent transcriptome under tight evolutionary constraint.

Language: Английский

Citations

0