Comparative Analysis of the Expression of Chondroitin Sulfate Subtypes and Their Inhibitory Effect on Axonal Growth in t
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ORIGINAL ARTICLE
Comparative Analysis of the Expression of Chondroitin Sulfate Subtypes and Their Inhibitory Effect on Axonal Growth in the Embryonic, Adult, and Injured Rat Brains Moon Hang Kim1 • So Ra Park2 • Byung Hyune Choi3
Received: 1 August 2020 / Revised: 1 August 2020 / Accepted: 16 August 2020 Ó The Korean Tissue Engineering and Regenerative Medicine Society 2020
Abstract BACKGROUND: Chondroitin sulfate glycosaminoglycans (CS-GAGs) are the primary inhibitory GAGs for neuronal growth after central nervous system (CNS) injury. However, the inhibitory or permissive activity of CS-GAG subtypes is controversial and depends on the physiological needs of CNS tissues. In this study, we investigated the characteristics and effects of CS-GAGs on axonal growth, which was isolated from the brain cortices of normal rat embryo at E18, normal adult rat brain and injured adult rat brain. METHODS: Isolated CS-GAGs from embryo, normal adult, and injured adult rat brains were used for analyzing their effect on attachment and axonal growth using modified spot assay with dorsal root ganglion (DRG) explants and cerebellar granule neurons (CGNs). CS-GAGs were separated using high performance liquid chromatography (HPLC), and the subtypes of CS-GAGs were analyzed. RESULTS: CS-GAGs of all three groups inhibited CGN attachment and axonal growth of DRGs. However, CS-GAGs of normal adult rat brain exhibited higher inhibitory activity than those of the other groups in both assays. When subtypes of CS-GAGs were analyzed using HPLC, CS-A (4S) was the most abundant in all three groups and found in largest amount in normal adult rat brain. In contrast, unsulfated CS (CS0) and CS-C (6S) were more abundant by 3–4-folds in E18 group than in the two adult groups. CONCLUSION: When compared with the normal adult rat brain, injured rat brain showed relatively similar patterns to that of embryonic rat brain at E18 in the expression of CS subtypes and their inhibitory effect on axonal growth. This phenomenon could be due to differential expression of CS-GAGs subtypes causing decrease in the amount of CS-A and mature-type CS proteoglycan core proteins. Keywords Chondroitin sulfate glycosaminoglycan Brain injury CNS development Chondroitin sulfate proteoglycan
1 Introduction & Byung Hyune Choi [email protected] 1
Biomedical Research Institute, Seoul National University Hospital, 101 Daehak-ro, Jongno-gu, Seoul 03080, Republic of Korea
2
Department of Physiology, Inha University College of Medicine, 100 Inha-ro, Michuhol-gu, Incheon 22212, Republic of Korea
3
Department of Biomedical Sciences, Inha University College of Medicine, 100 Inha-ro, Michuhol-gu, Incheon 22212, Republic of Korea
Chondroitin sulfate proteoglycans (CSPGs) are the most abundant proteoglycans in the central nervous system (CNS). CSPGs are secreted into the extracellular matrix (ECM) and are responsible for various regulatory functions during development, adulthood, and injury [1]. During mice development, juvenile-type CSPGs such as neuroca
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