Journal article
Targeted downregulation of N-acetylgalactosamine 4-sulfate 6-O-sulfotransferase (GalNAc4S6ST) significantly mitigates chondroitin sulfate proteoglycan (CSPG) mediated inhibition
Glia, v 59(6), pp 981-996
Jun 2011
PMID: 21456043
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Chondroitin Sulfate-4,6 (CS-E) glycosaminoglycan (GAG) upregulation in astroglial scars is a major contributor to CS proteoglycan (CSPG) mediated inhibition (
Gilbert et al. 2005
). However, the role of N-acetylgalactosamine 4-sulfate 6-O-sulfotransferase (GalNAc4S6ST) catalyzed sulfation of CS-E, and its contribution to CSPG mediated inhibition of CNS regeneration remains to be fully elucidated. Here, we used
in situ
hybridization to show localized upregulation of GalNAc4S6ST mRNA after CNS injury. Using
in vitro
spot assays with immobilized CS-E, we demonstrate dose dependent inhibition of rat embryonic day 18 (E18) cortical neurons. To determine whether selective downregulation of CS-E affected the overall inhibitory character of extracellular matrix produced by reactive astrocytes, single [against (chondroitin 4) sulfotransferase 11 (C4ST1) or GalNAc4S6ST mRNA) or double (against C4ST1 and GalNAc4S6ST mRNA] siRNA treatments were conducted and assayed using quantitative real-time PCR (qRT-PCR) and high performance liquid chromatography (HPLC) to confirm the specific downregulation of CS-4S GAG (CS-A) and CS-E. Spot and Bonhoeffer stripe assays using astrocyte conditioned media (ACM) from siRNA treated rat astrocytes showed a significant decrease in inhibition of neuronal attachment and neurite extensions when compared to untreated and TGFα treated astrocytes. These findings reveal that selective attenuation of CS-E via siRNA targeting of GalNAc4S6ST significantly mitigates CSPG mediated inhibition of neurons, potentially offering a novel intervention strategy for CNS injury.
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Details
- Title
- Targeted downregulation of N-acetylgalactosamine 4-sulfate 6-O-sulfotransferase (GalNAc4S6ST) significantly mitigates chondroitin sulfate proteoglycan (CSPG) mediated inhibition
- Creators
- Lohitash Karumbaiah - Wallace H. Coulter Department of Biomedical Engineering at Georgia Institute of Technology and Emory University School of Medicine, Atlanta, GA 30332Sanjay Anand - Wallace H. Coulter Department of Biomedical Engineering at Georgia Institute of Technology and Emory University School of Medicine, Atlanta, GA 30332Rupal Thazhath - Department of Biology, Georgia Institute of Technology, Atlanta, GA 30332Yinghui Zhong - Wallace H. Coulter Department of Biomedical Engineering at Georgia Institute of Technology and Emory University School of Medicine, Atlanta, GA 30332Robert J McKeon - Department of Cell Biology, Emory University, GA 30322Ravi V Bellamkonda - Wallace H. Coulter Department of Biomedical Engineering at Georgia Institute of Technology and Emory University School of Medicine, Atlanta, GA 30332
- Publication Details
- Glia, v 59(6), pp 981-996
- Publisher
- Wiley
- Grant note
- R01 NS043486-05 || NS / National Institute of Neurological Disorders and Stroke : NINDS
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- School of Biomedical Engineering, Science, and Health Systems
- Web of Science ID
- WOS:000289674400012
- Scopus ID
- 2-s2.0-79954456797
- Other Identifier
- 991014877926304721
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- Collaboration types
- Domestic collaboration
- Web of Science research areas
- Neurosciences