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Characterization of Glycosaminoglycan Disaccharide Composition in Astrocyte Primary Cultures and the Cortex of Neonatal Rats

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Abstract

Astrocytes are major producers of the extracellular matrix (ECM), which is involved in the plasticity of the developing brain. In utero alcohol exposure alters neuronal plasticity. Glycosaminoglycans (GAGs) are a family of polysaccharides present in the extracellular space; chondroitin sulfate (CS)- and heparan sulfate (HS)-GAGs are covalently bound to core proteins to form proteoglycans (PGs). Hyaluronic acid (HA)-GAGs are not bound to core proteins. In this study we investigated the contribution of astrocytes to CS-, HS-, and HA-GAG production by comparing the makeup of these GAGs in cortical astrocyte cultures and the neonatal rat cortex. We also explored alterations induced by ethanol in GAG and core protein levels in astrocytes. Finally, we investigated the relative expression in astrocytes of CS-PGs of the lectican family of proteins, major components of the brain ECM, in vivo using translating ribosome affinity purification (TRAP) (in Aldh1l1-EGFP-Rpl10a mice. Cortical astrocytes produce low levels of HA and show low expression of genes involved in HA biosynthesis compared to the whole developing cortex. Astrocytes have high levels of chondroitin-0-sulfate (C0S)-GAGs (possibly because of a higher sulfatase enzyme expression) and HS-GAGs. Ethanol upregulates C4S-GAGs as well as brain-specific lecticans neurocan and brevican, which are highly enriched in astrocytes of the developing cortex in vivo. These results begin to elucidate the role of astrocytes in the biosynthesis of CS- HS- and HA-GAGs, and suggest that ethanol-induced alterations of neuronal development may be in part mediated by increased astrocyte GAG levels and neurocan and brevican expression.

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The authors confirm that the data support their published claims and comply with field standards.

Abbreviations

AMAC:

2-Aminoacridone

ARSB:

Arylsulfatase B

CS:

Chondroitin Sulfate

ECM:

Extracellular Matrix

FASD:

Fetal Alcohol Spectrum Disorders

GAGs:

Glycosaminoglycans

GALNS:

Galactosamine (N-acetyl)-6 Sulfatase

HA:

Hyaluronic Acid

HS:

Heparan Sulfate

NDST:

N-Deaceylase/N-sulfotransferases

PD:

Postnatal Day

PGs:

Proteoglycans

RRID:

Research Resource Identifier

SUMF1:

Sulfatase Modifying Factor 1

TRAP:

Translating ribosome affinity purification

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Acknowledgement

We thank Mr. Nicholas Margolies for the critical review of the manuscript and Ms. Shelley H. Bloom and Dr. Clare J. Wilhelm for their technical assistance with astrocyte cultures and treatments.

Funding

This work was supported by the Department of Veterans Affairs Merit Review BX001819, and National Institutes of Health R01AA022948 (MG) and the New York State Spinal Cord Injury Research Board (SCRIB) Grant DOH01-PART2-2017 (RJL).

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Authors and Affiliations

  1. Department of Behavioral Neuroscience, Oregon Health & Science University, Portland, OR, USA

    Xiaolu Zhang, Joel G. Hashimoto & Marina Guizzetti

  2. VA Portland Health Care System, R&D39, 3710 SW Veterans Hospital Road, Portland, OR, 97239, USA

    Xiaolu Zhang, Joel G. Hashimoto & Marina Guizzetti

  3. Chemistry and Chemical Biology, Biomedical Engineering, Chemical and Biological Engineering, and Biology; Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY, USA

    Xiaorui Han, Fuming Zhang & Robert J. Linhardt

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  1. Xiaolu Zhang
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  2. Joel G. Hashimoto
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  3. Xiaorui Han
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  4. Fuming Zhang
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  6. Marina Guizzetti
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Contributions

MG conceived the study; MG, XZ, and RJL designed experiments. Material preparation, data collection and analysis were performed by XZ, JGH, XH and FZ; MG and XZ wrote the paper with suggestions from JGH and RJL. All authors read and approved the final manuscript.

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Correspondence to Xiaolu Zhang or Marina Guizzetti.

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Zhang, X., Hashimoto, J.G., Han, X. et al. Characterization of Glycosaminoglycan Disaccharide Composition in Astrocyte Primary Cultures and the Cortex of Neonatal Rats. Neurochem Res 46, 595–610 (2021). https://doi.org/10.1007/s11064-020-03195-9

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