Chondroitin sulfate-binding peptides block chondroitin 6-sulfate inhibition of cortical neurite growth

Butterfield, Karen Chao; Conovaloff, Aaron W.; Caplan, Michael R.; Panitch, Alyssa

doi:10.1016/j.neulet.2010.04.070

Cited by 28 publications

(21 citation statements)

References 29 publications

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“…Additional experiments such as rescue studies validating the role of C-6-S would be valuable but currently C-6-S is not commercially available. Both C-6-S and C-4-S disaccharides have been shown to inhibit neurite growth from embryonic neurons in cell culture however, their role with respect to adult neurons, such as those that innervate the adult IVD, are less clear [44, 45]. These studies suggest that free intact GAGs such as CS side-chains could be effective at inhibiting neurite growth and this is supported by work demonstrating that recombinant aggrecan depleted of chondroitin sulfated side-chains was unable to inhibit neurite extension [46].…”

Section: Discussionmentioning

confidence: 99%

Intact glycosaminoglycans from intervertebral disc-derived notochordal cell-conditioned media inhibit neurite growth while maintaining neuronal cell viability

et al. 2015

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BACKGROUND CONTEXT Painful human intervertebral discs (IVDs) exhibit nerve growth deep into the IVD. Current treatments for discogenic back pain do not address the underlying mechanisms propagating pain and are often highly invasive or only offer temporary symptom relief. The notochord produces factors during development that pattern the spine and inhibit growth of dorsal root ganglia (DRG) axons into the IVD. We hypothesize that notochordal cell (NC) conditioned medium (NCCM) includes soluble factors capable of inhibiting neurite growth and may represent a future therapeutic target. PURPOSE To test if NCCM can inhibit neurite growth and to determine if NC-derived Glycosaminoglycans (GAGs) are necessary candidates for this inhibition. STUDY DESIGN Human neuroblastoma (SH-SY5Y) cells and rat dorsal root ganglion (DRGs) cells were treated with NCCM in 2D culture in vitro and digestion and mechanistic studies determined if specific GAGs were responsible for inhibitory effects. METHODS NCCM was generated from porcine nucleus pulposus tissue that was cultured in DMEM for 4 days. A dose study was performed using SH-SY5Y cells that were seeded in basal medium for 24hrs and neurite outgrowth and cell viability were assessed following treatment with basal or NCCM (10% and 100%) media for 48hrs. GAGs from NCCM were characterized using multiple digestions and liquid chromatography mass spectroscopy (LC-MS). Neurite growth was assessed on both SH-SY5Y and DRG cells following treatment with NCCM with and without GAG digestion. RESULTS NCCM significantly inhibited neurite outgrowth from SH-SY5Y cells compared to Basal controls without dose or cytotoxic effects; % neurite expressing cells were 39.0±2.9, 27.3±3.6 & 30.2±2.7, and mean neurite length was 60.3±3.5μm, 50.8±2.4μm, 53.2±3.7μm for Basal, 10% NCCM & 100% NCCM, respectively. Digestions and LC-MS determined that Chondroitin-6-Sulfate was the major GAG chain in NCCM. Neurite growth from SH-SY5Y and DRG cells was not inhibited when cells were treated with NCCM with digested chondroitin sulfate (CS). CONCLUSIONS Soluble factors derived from NCCM were capable of inhibiting neurite outgrowth in multiple neural cell types without any negative effects on cell viability. Cleavage of GAGs via digestion was necessary to reverse the neurite inhibition capacity of NCCM. We conclude that intact GAGs such as CS secreted from NCs are potential candidates that could be useful to reduce neurite growth in painful IVDs.

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Section: Discussionmentioning

confidence: 99%

Intact glycosaminoglycans from intervertebral disc-derived notochordal cell-conditioned media inhibit neurite growth while maintaining neuronal cell viability

et al. 2015

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“…Although much evidence has accumulated suggesting that it is the GAG chain moieties that are recognized by nerve growth cones, fewer studies have specifically addressed the differential effects that individual GAG moities have on a particular nerve cell. 13,20,22,46,47,100,101 Here, we used a modified spot assay designed to mimic an in vivo environment whereby GAGs are immobilized and presented solely to nerve growth cones, but not to their neuronal cell bodies, similar to how GAGs would be encountered by growth cones in a developing embryo. Our results show that each GAG tested in the study displayed its own unique potential for inhibiting trigeminal nerve growth cone migration in vitro.…”

Section: Discussionmentioning

confidence: 99%

Corneal Sulfated Glycosaminoglycans and Their Effects on Trigeminal Nerve Growth Cone Behavior In Vitro: Roles for ECM in Cornea Innervation

Schwend

Deaton

Zhang

et al. 2012

Invest. Ophthalmol. Vis. Sci.

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“…Disaccharide analysis of rat brain and spinal cord CSPGs also revealed that CS-E content increased by 20% only when detergent extracted (Deepa et al 2006), indicating that CS-E is likely attached to the perineural net (PNN) associated isoforms of neurocan, brevican, aggrecan and versican. Recently, sulfation patterns of CS-disaccharides have been the subject of much scrutiny (Butterfield et al 2010; Gilbert et al 2005; Ito et al 2010; Properzi et al 2005; Wang et al 2008). Properzi et al (2005) reported that the inhibitory properties of proteoglycans upregulated after CNS injury were mediated by CS-C.…”

Section: Discussionmentioning

confidence: 99%

Targeted downregulation of N‐acetylgalactosamine 4‐sulfate 6‐O‐sulfotransferase significantly mitigates chondroitin sulfate proteoglycan‐mediated inhibition

Karumbaiah

Anand

Thazhath

et al. 2011

Glia

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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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Chondroitin sulfate-binding peptides block chondroitin 6-sulfate inhibition of cortical neurite growth

Cited by 28 publications

References 29 publications

Intact glycosaminoglycans from intervertebral disc-derived notochordal cell-conditioned media inhibit neurite growth while maintaining neuronal cell viability

Intact glycosaminoglycans from intervertebral disc-derived notochordal cell-conditioned media inhibit neurite growth while maintaining neuronal cell viability

Corneal Sulfated Glycosaminoglycans and Their Effects on Trigeminal Nerve Growth Cone Behavior In Vitro: Roles for ECM in Cornea Innervation

Targeted downregulation of N‐acetylgalactosamine 4‐sulfate 6‐O‐sulfotransferase significantly mitigates chondroitin sulfate proteoglycan‐mediated inhibition

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