ABSTRACT
Lumican is a keratan sulfate proteoglycan that belongs to the small leucine-rich proteoglycan family. Research has lifted the veil on the versatile roles of lumican in the pathogenesis of eye diseases. Lumican has pivotal roles in the maintenance of physiological tissue homogenesis and is often upregulated in pathological conditions, e.g., fibrosis, scar tissue formation in injured tissues, persistent inflammatory responses and immune anomaly, etc. Herein, we will review literature regarding the role of lumican in pathogenesis of inherited congenital and acquired eye diseases, e.g., cornea dystrophy, cataract, glaucoma and chorioretinal diseases, etc.
Subject(s)
Eye Diseases , Lumican , Humans , Chondroitin Sulfate Proteoglycans/physiology , Cornea/pathology , Eye Diseases/metabolism , Eye Diseases/pathology , Keratan Sulfate/physiology , Proteoglycans/physiologyABSTRACT
Composition of the brain extracellular matrix changes in time as maturation proceeds. Chondroitin sulfate proteoglycan 5 (CSPG-5), also known as neuroglycan C, has been previously associated to differentiation since it shapes neurite growth and synapse forming. Here, we show that this proteoglycan persists in the postnatal rat brain, and its expression is higher in cortical regions with plastic properties, including hippocampus and the medial prefrontal cortex at the end of the second postnatal week. Progressively accumulating after birth, CSPG-5 typically concentrates around glutamatergic and GABAergic terminals in twelve-week old rat hippocampus. CSPG-5-containing perisynaptic matrix rings often appear at the peripheral margin of perineuronal nets. Electron microscopy and analysis of synaptosomal fraction showed that CSPG-5 accumulates around, and is associated to synapses, respectively. In vitro analyses suggest that neurons, but less so astrocytes, express CSPG-5 in rat primary neocortical cultures, and CSPG-5 produced by transfected neuroblastoma cells appear at endings and contact points of neurites. In human subjects, CSPG-5 expression shifts in brain areas of the default mode network of suicide victims, which may reflect an impact in the pathogenesis of psychiatric diseases or support diagnostic power.
Subject(s)
Cerebellar Cortex/metabolism , Chondroitin Sulfate Proteoglycans/physiology , Membrane Proteins/physiology , Neurites/metabolism , Proteoglycans/physiology , Synapses/metabolism , Animals , Cell Line , Humans , Male , Rats , Rats, WistarABSTRACT
Chondroitin sulfate (CS), a type of glycosaminoglycan (GAG), is a linear acidic polysaccharide comprised of repeating disaccharides, modified with sulfate groups at various positions. Except for hyaluronan (HA), GAGs are covalently bound to core proteins, forming proteoglycans (PGs). With highly negative charges, GAGs interact with a variety of physiologically active molecules, including cytokines, chemokines, and growth factors, and control cell behavior during development and in the progression of diseases, including cancer, infections, and inflammation. Heparan sulfate (HS), another type of GAG, and HA are well reported as regulators for leukocyte migration at sites of inflammation. There have been many reports on the regulation of immune cell function by HS and HA; however, regulation of immune cells by CS has not yet been fully understood. This article focuses on the regulatory function of CS in antigen-presenting cells, including macrophages and dendritic cells, and refers to CSPGs, such as versican and biglycan, and the cell surface proteoglycan, syndecan.
Subject(s)
Adaptive Immunity , Antigen-Presenting Cells/drug effects , Chondroitin Sulfate Proteoglycans/physiology , Chondroitin Sulfates/physiology , Dendritic Cells/drug effects , Immunity, Innate , Macrophages/drug effects , Antigen-Presenting Cells/immunology , Biglycan/physiology , Carbohydrate Conformation , Carbohydrate Sequence , Chondroitin Sulfate Proteoglycans/pharmacology , Chondroitin Sulfates/pharmacology , Dendritic Cells/immunology , Humans , Hyaluronan Receptors/physiology , Macrophages/immunology , Receptor-Like Protein Tyrosine Phosphatases, Class 2/physiology , Structure-Activity Relationship , Syndecans/physiology , Toll-Like Receptors/physiology , Versicans/physiologyABSTRACT
Proteoglycans, a class of carbohydrate-modified proteins, often modulate growth factor signaling on the cell surface. However, the molecular mechanism by which proteoglycans regulate signal transduction is largely unknown. In this study, using a recently developed glycoproteomic method, we found that Windpipe (Wdp) is a novel chondroitin sulfate proteoglycan (CSPG) in Drosophila. Wdp is a single-pass transmembrane protein with leucin-rich repeat (LRR) motifs and bears three CS sugar chain attachment sites in the extracellular domain. Here we show that Wdp modulates the Hedgehog (Hh) pathway. In the wing disc, overexpression of wdp inhibits Hh signaling, which is dependent on its CS chains and the LRR motifs. The wdp null mutant flies show a specific defect (supernumerary scutellar bristles) known to be caused by Hh overexpression. RNA interference knockdown and mutant clone analyses showed that loss of wdp leads to the up-regulation of Hh signaling. Altogether, our study demonstrates a novel role of CSPGs in regulating Hh signaling.
Subject(s)
Chondroitin Sulfate Proteoglycans/physiology , Drosophila Proteins/physiology , Drosophila melanogaster/physiology , Hedgehog Proteins/physiology , Membrane Proteins/physiology , Amino Acid Sequence , Animals , Animals, Genetically Modified , Chondroitin Sulfates/metabolism , Drosophila Proteins/genetics , Drosophila melanogaster/genetics , Drosophila melanogaster/growth & development , Female , Gene Expression Regulation, Developmental , Gene Knockdown Techniques , Heparitin Sulfate/metabolism , Imaginal Discs/metabolism , Larva , Membrane Proteins/genetics , RNA Interference , RNA, Small Interfering/genetics , RNA, Small Interfering/pharmacology , Signal Transduction/physiology , Wings, Animal/growth & development , Wings, Animal/ultrastructureABSTRACT
Dermatopontin (DPT) is an extensively distributed non-collagenous component of the extracellular matrix predominantly found in the dermis of the skin, and consequently expressed in several tissues. In this study, we explored the role of DPT in myogenesis and perceived that it enhances the cell adhesion, reduces the cell proliferation and promotes the myoblast differentiation in C2C12 cells. Our results reveal an inhibitory effect with fibronectin (FN) in myoblast differentiation. We also observed that DPT and fibromodulin (FMOD) regulate positively to each other and promote myogenic differentiation. We further predicted the 3D structure of DPT, which is as yet unknown, and validated it using state-of-the-art in silico tools. Furthermore, we explored the in-silico protein-protein interaction between DPT-FMOD, DPT-FN, and FMOD-FN, and perceived that the interaction between FMOD-FN is more robust than DPT-FMOD and DPT-FN. Taken together, our findings have determined the role of DPT at different stages of the myogenic process.
Subject(s)
Chondroitin Sulfate Proteoglycans , Extracellular Matrix Proteins , Fibromodulin , Fibronectins , Muscle Development/physiology , Muscle, Skeletal/metabolism , Animals , Cell Adhesion/physiology , Cell Differentiation/physiology , Cell Line , Cell Proliferation/physiology , Chondroitin Sulfate Proteoglycans/chemistry , Chondroitin Sulfate Proteoglycans/physiology , Extracellular Matrix Proteins/chemistry , Extracellular Matrix Proteins/physiology , Fibromodulin/chemistry , Fibromodulin/metabolism , Fibronectins/chemistry , Fibronectins/metabolism , Mice , Protein BindingABSTRACT
Chondroitin sulfate proteoglycan (CSPG) is a candidate regulator of embryonic neurogenesis. The aim of this study was to specify the functional significance of CSPG in adult hippocampal neurogenesis using male mice. Here, we showed that neural stem cells and neuronal progenitors in the dentate gyrus were covered in part by CSPG. Pharmacological depletion of CSPG in the dentate gyrus reduced the densities of neuronal progenitors and newborn granule cells. 3D reconstruction of newborn granule cells showed that their maturation was inhibited by CSPG digestion. The novel object recognition test revealed that CSPG digestion caused cognitive memory impairment. Western blot analysis showed that expression of ß-catenin in the dentate gyrus was decreased by CSPG digestion. The amount of CSPG in the dentate gyrus was increased by enriched environment (EE) and was decreased by forced swim stress. In addition, EE accelerated the recovery of CSPG expression in the dentate gyrus from the pharmacological depletion and promoted the restoration of granule cell production. Conversely, the densities of newborn granule cells were also decreased in mice that lacked chondroitin sulfate N-acetylgalactosaminyltransferase 1 (CSGalNAcT1), a key enzyme for CSPG synthesis (T1KO mice). The capacity of EE to promote granule cell production and improve cognitive memory was impaired in T1KO mice. These findings indicate that CSPG is involved in the regulation of adult hippocampal neurogenesis and suggest that increased synthesis of CSPG by CSGalNacT1 may mediate promotion of granule cell production and improvement of cognitive memory in response to EE.SIGNIFICANCE STATEMENT Chondroitin sulfate proteoglycan (CSPG) is a candidate regulator of embryonic neurogenesis. Here, we specified the role of CSPG in adult neurogenesis in the mouse hippocampus. Digestion of CSPG in the dentate gyrus impaired granule cell production and cognitive memory. Enriched environment (EE) promoted the recovery of CSPG expression and granule cell production from the CSPG digestion. Additionally, adult neurogenesis was impaired in mice that lacked a key enzyme for CSPG synthesis (T1KO mice). The capacity of EE to promote granule cell production and cognitive memory was impaired in T1KO mice. Altogether, these findings indicate that CSPG underlies adult hippocampal neurogenesis and suggest that increased synthesis of CSPG may mediate promotion of granule cell production in response to EE.
Subject(s)
Chondroitin Sulfate Proteoglycans/physiology , Environment , Hippocampus/physiology , Neurogenesis , Neurons/physiology , Animals , Cognition/physiology , Hippocampus/cytology , Male , Memory/physiology , Mice, Inbred C57BL , Mice, Knockout , N-Acetylgalactosaminyltransferases/genetics , Neural Stem Cells/cytology , Neural Stem Cells/physiology , Neurons/cytology , Recognition, Psychology/physiologyABSTRACT
CSPGs are components of the extracellular matrix in the nervous system, where they serve as cues for axon guidance during development. After a peripheral nerve injury, CSPGs switch roles and become axon inhibitors and become diffusely distributed at the injury site. To investigate whether the spatial distribution of CSPGs affects their role, we combined in vitro DRG cultures with CSPG stripe or coverage assays to simulate the effect of a patterned substrate or dispersive distribution of CSPGs on growing neurites. We observed neurite steering at linear CSPG interfaces and neurite inhibition when diffused CSPGs covered the distal but not the proximal segment of the neurite. The repellent and inhibitory effects of CSPGs on neurite outgrowth were associated with the disappearance of focal actin filaments on growth cones. The application of an actin polymerization inducer, jasplakinolide, allowed neurites to break through the CSPG boundary and grow on CSPG-coated surfaces. The results of our study collectively reveal a novel mechanism that explains how the spatial distribution of CSPGs determines whether they act as a cue for axon guidance or as an axon-inhibiting factor. Increasing our understanding of this issue may promote the development of novel therapeutic strategies that regulate the spatial distributions of CSPGs to use them as an axon guidance cue.
Subject(s)
Actin Cytoskeleton/physiology , Chondroitin Sulfate Proteoglycans/physiology , Ganglia, Spinal/physiology , Nerve Regeneration/physiology , Signal Transduction/physiology , Actin Cytoskeleton/drug effects , Animals , Cells, Cultured , Depsipeptides/pharmacology , Dose-Response Relationship, Drug , Ganglia, Spinal/cytology , Ganglia, Spinal/drug effects , Nerve Regeneration/drug effects , Rats , Rats, Sprague-Dawley , Signal Transduction/drug effectsABSTRACT
The inability to recover functions lost after severe spinal cord injury has been recognized for millennia and was first attributed to a failure of spinal cord neural regeneration over 100 years ago. The last forty years have seen intense research into achieving such regeneration, but in spite of conceptual advances and many reports announcing successful interventions, progress has been slow and often controversial. Here, I examine consequential advances and setbacks, and critically consider assumptions underlying certain approaches. I argue that expanding mechanistic knowledge about multiple forms of neural regeneration, why they fail and how they can restore function will resolve conceptual contentions and push the field forward.
Subject(s)
Spinal Cord Regeneration/physiology , Animals , Astrocytes/pathology , Axons/pathology , Axons/physiology , Chondroitin Sulfate Proteoglycans/physiology , Gray Matter/physiology , Humans , Myelin Sheath/physiology , Neural Pathways/physiology , Neuroglia/pathologyABSTRACT
A subset of monoclonal anti-DNA autoantibodies enters a variety of living cells. Here, we aimed to identify the endocytic receptors recognized by an internalizing anti-nucleic acid autoantibody, the 3D8 single-chain variable fragment (scFv). We found that cell surface binding and internalization of 3D8 scFv were inhibited markedly in soluble heparan sulfate (HS)/chondroitin sulfate (CS)-deficient or -removed cells and in the presence of soluble HS and CS. 3D8 scFv colocalized intracellularly with either HS proteoglycans (HSPGs) or CSPGs in HeLa cells. 3D8 scFv was co-endocytosed and co-precipitated with representative individual HSPG and CSPG molecules: syndecan-2 (a transmembrane HSPG), glypican-3 (a glycosylphosphatidylinositol (GPI)-anchored HSPG); CD44 (a transmembrane CSPG); and brevican (a GPI-anchored CSPG). Collected data indicate that 3D8 scFv binds to the negatively charged sugar chains of both HSPGs and CSPGs and is then internalized along with these molecules, irrespective of how these proteoglycans are associated with the cell membrane. This is the first study to show that anti-DNA antibodies enter cells via both HSPGs and CSPGs simultaneously. The data may aid understanding of endocytic receptors that bind anti-DNA autoantibodies. The study also provides insight into potential cell membrane targets for macromolecular delivery.
Subject(s)
Chondroitin Sulfate Proteoglycans/physiology , Heparan Sulfate Proteoglycans/physiology , Animals , Antibodies, Antinuclear/physiology , CD13 Antigens/immunology , Cell Membrane/metabolism , Chondroitin Sulfate Proteoglycans/metabolism , Chondroitin Sulfates/metabolism , Cytoplasm/metabolism , Endocytosis/physiology , Glycosaminoglycans/metabolism , Glypicans/immunology , HeLa Cells , Heparan Sulfate Proteoglycans/metabolism , Heparitin Sulfate/metabolism , Humans , Hyaluronan Receptors/immunology , Nucleic Acids/metabolism , Transport VesiclesABSTRACT
Abnormal epigenetic regulation can cause the nervous system to develop abnormally. Here, we sought to understand the mechanism by which this occurs by investigating the protein complex cohesin, which is considered to regulate gene expression and, when defective, is associated with higher-level brain dysfunction and the developmental disorder Cornelia de Lange syndrome (CdLS). We generated conditional Smc3-knockout mice and observed greater dendritic complexity and larger numbers of immature synapses in the cerebral cortex of Smc3+/- mice. Smc3+/- mice also exhibited more anxiety-related behavior, which is a symptom of CdLS. Further, a gene ontology analysis after RNA-sequencing suggested the enrichment of immune processes, particularly the response to interferons, in the Smc3+/- mice. Indeed, fewer synapses formed in their cortical neurons, and this phenotype was rescued by STAT1 knockdown. Thus, low levels of cohesin expression in the developing brain lead to changes in gene expression that in turn lead to a specific and abnormal neuronal and behavioral phenotype.
Subject(s)
Anxiety/etiology , Brain/physiopathology , Cell Cycle Proteins/deficiency , Chromosomal Proteins, Non-Histone/deficiency , Synapses/physiology , Animals , Anxiety/physiopathology , Brain/metabolism , Brain Chemistry/physiology , Cell Cycle Proteins/physiology , Chondroitin Sulfate Proteoglycans/physiology , Chromosomal Proteins, Non-Histone/physiology , Female , Gene Expression Regulation/physiology , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , CohesinsABSTRACT
Many current peripheral nerve repair strategies focus on delivering positive, growth promoting cues (e.g. extracellular matrix, ECM) while eliminating negative, growth inhibiting cues (e.g. chondroitin sulfate proteoglycans, CSPGs) at the injury site. We hypothesized that recapitulating the positive and negative cues of the peripheral nerve injury microenvironment would improve regeneration. First, we tested the effects of a characteristic CSPG, chondroitin sulfate A (CSA) on neurite dynamics of dissociated chick embryo dorsal root ganglion (DRG) neurons using time lapse video microscopy. DRG growth was recorded on different adhesive substrates, including a novel, porcine-derived spinal cord matrix (SCM). The SCM significantly increased frequency of neurite extension coordinated by a significant reduction in the neurites' time spent stalled. The SCM also mitigated inhibitory effects of CSA, producing longer neurites than the controls without CSA treatment. Next we aimed to elucidate receptors involved in mediating this behavior by testing the ability of CSA to upregulate cell-substrate binding receptors using flow cytometry. Our results showed a significant increase in syndecan-3 receptor expression in neurons treated with CSA. Furthermore, syndecans would most likely bind to the sulfated glycosaminoglycans measured in the SCM. Finally, we evaluated neurite growth on biomaterial scaffolds featuring CSA and SCM cues. Our results showed significantly increased neurite outgrowth on electrospun hyaluronic acid fibers with SCM and low levels of CSA. Higher incorporation of CSA maintained its inhibitory properties. Future work will evaluate coupling CSPGs with growth-permissive ECM to assess the combined effect on neurite outgrowth.
Subject(s)
Nerve Regeneration/physiology , Neurites/physiology , Animals , Biocompatible Materials/chemistry , Cellular Microenvironment/physiology , Chick Embryo , Chondroitin Sulfate Proteoglycans/physiology , Chondroitin Sulfates/physiology , Extracellular Matrix/physiology , Ganglia, Spinal/cytology , Materials Testing , Nanofibers/chemistry , Neurites/ultrastructure , Peripheral Nerve Injuries/physiopathology , Peripheral Nerve Injuries/therapy , Spinal Cord/physiology , Swine , Syndecan-3/physiology , Time-Lapse Imaging , Tissue Engineering/methods , Tissue Scaffolds/chemistryABSTRACT
The hypothalamus controls metabolism, stress responses, and instinctive behaviors for individual survival and species preservation. Recent studies suggest that hypothalamic neurons retain plasticity throughout adulthood, which enables these neurons to respond to various kinds of changes in environment, nutrients, and fluctuating hormones. One of the mechanisms underlying the regulation of neural plasticity is the formation of a stable extracellular matrix (ECM) structure called perineuronal nets (PNNs). PNNs are large aggregates of heterogeneous ECM molecules such as chondroitin sulfate proteoglycans (CSPGs), hyaluronan, their link proteins, and tenascin-R. PNNs surround the cell body and proximal dendrites of a subset of neurons and limit adult neural plasticity. This review describes the CSPG-based ECM, including the PNNs, with a special focus on the hypothalamus of mice. We first provide an overview of PNNs in terms of their structure, molecular components, and functions, most of which have been demonstrated by extrahypothalamic studies. Second, we show the presence or absence of PNNs within individual hypothalamic regions and then describe non-PNN-formed ECM containing CSPGs that can be observed in particular hypothalamic regions. Finally, we will introduce a newly identified mouse hypothalamic area that we named the perifornical area of the anterior hypothalamus (PeFAH), which contains a cluster of PNN-positive neurons. PeFAH neurons express enkephalin and have bidirectional connections with the lateral septum. The anterior hypothalamus and lateral septum are thought to regulate defensive behaviors; therefore, the PeFAH neurons and PNNs around them could be involved in the regulation of defensive behaviors.
Subject(s)
Extracellular Matrix/metabolism , Extracellular Matrix/physiology , Hypothalamus, Anterior/cytology , Hypothalamus, Anterior/physiology , Neurons/metabolism , Neurons/physiology , Animals , Behavior, Animal , Chondroitin Sulfate Proteoglycans/metabolism , Chondroitin Sulfate Proteoglycans/physiology , Chondroitin Sulfate Proteoglycans/ultrastructure , Defense Mechanisms , Enkephalins/metabolism , Extracellular Matrix/ultrastructure , Hypothalamus, Anterior/metabolism , Mice, Inbred C57BL , Neuronal PlasticityABSTRACT
CSPG4/NG2 is a multifunctional transmembrane protein with limited distribution in adult tissues including articular cartilage. The purpose of this study was to investigate the possible roles of CSPG4/NG2 in chondrosarcomas and to establish whether this molecule may have potential for targeted therapy. Stable knock-down of CSPG4/NG2 in the JJ012 chondrosarcoma cell line by shRNA resulted in decreased cell proliferation and migration as well as a decrease in gene expression of the MMP (matrix metalloproteinase) 3 protease and ADAMTS4 (aggrecanase). Chondrosarcoma cells in which CSPG4/NG2 was knocked down were more sensitive to doxorubicin than wild-type cells. The results indicate that CSPG4/NG2 has roles in regulating chondrosarcoma cell function in relation to growth, spread and resistance to chemotherapy and that anti-CSPG4/NG2 therapies may have potential in the treatment of surgically unresectable chondrosarcoma.
Subject(s)
Bone Neoplasms/pathology , Chondroitin Sulfate Proteoglycans/physiology , Chondrosarcoma/pathology , Membrane Proteins/physiology , Adult , Aged , Antineoplastic Agents/pharmacology , Bone Neoplasms/genetics , Bone Neoplasms/metabolism , Cartilage, Articular/enzymology , Cell Adhesion/physiology , Cell Death/drug effects , Cell Line, Tumor , Cell Movement/physiology , Cell Proliferation/physiology , Chondroitin Sulfate Proteoglycans/genetics , Chondroitin Sulfate Proteoglycans/metabolism , Chondrosarcoma/genetics , Chondrosarcoma/metabolism , Docetaxel , Doxorubicin/pharmacology , Drug Resistance, Neoplasm/genetics , Female , Gene Expression Regulation, Neoplastic , Gene Knockdown Techniques/methods , Humans , Male , Membrane Proteins/genetics , Membrane Proteins/metabolism , Middle Aged , Neoplasm Invasiveness , Neoplasm Proteins/genetics , Neoplasm Proteins/metabolism , Neoplasm Proteins/physiology , Peptide Hydrolases/metabolism , RNA, Small Interfering/genetics , Taxoids/pharmacology , Young AdultABSTRACT
The presence of cancer stem cells (CSCs) or tumor-initiating cells can lead to cancer recurrence in a permissive cell-microenvironment interplay, promoting invasion in glioblastoma (GBM) and neuroblastoma (NB). Extracellular matrix (ECM) small leucine-rich proteoglycans (SLRPs) play multiple roles in tissue homeostasis by remodeling the extracellular matrix (ECM) components and modulating intracellular signaling pathways. Due to their pan-inhibitory properties against receptor tyrosine kinases (RTKs), SLRPs are reported to exert anticancer effects in vitro and in vivo. However, their roles seem to be tissue-specific and they are also involved in cancer cell migration and drug resistance, paving the way to complex different scenarios. The aim of this study was to determine whether the SLRPs decorin (DCN) and lumican (LUM) are recruited in cell plasticity and microenvironmental adaptation of differentiated cancer cells induced towards stem-like phenotype. Floating neurospheres were generated by applying CSC enrichment medium (neural stem cell serum-free medium, NSC SFM) to the established SF-268 and SK-N-SH cancer cell lines, cellular models of GBM and NB, respectively. In both models, the time-dependent synergistic activation of DCN and LUM was observed. The highest DCN and LUM mRNA/protein expression was detected after cell exposure to NSC SFM for 8/12 days, considering these cells as SLRP-expressing (SLRP+) CSC-like. Ultrastructural imaging showed the cellular heterogeneity of both the GBM and NB neurospheres and identified the inner living cells. Parental cell lines of both GBM and NB grew only in soft agar + NSC SFM, whereas the secondary neurospheres (originated from SLRP+ t8 CSC-like) showed lower proliferation rates than primary neurospheres. Interestingly, the SLRP+ CSC-like from the GBM and NB neurospheres were resistant to temozolomide (TMZ) at concentrations >750 µM. Our results suggest that GBM and NB CSC-like promote the activation of huge quantities of SLRP in response to CSC enrichment, simultaneously acquiring TMZ resistance, cellular heterogeneity, and a quiescent phenotype, suggesting a novel pivotal role for SLRP in drug resistance and cell plasticity of CSC-like, allowing cell survival and ECM/niche modulation potential.
Subject(s)
Brain Neoplasms/pathology , Chondroitin Sulfate Proteoglycans/physiology , Dacarbazine/analogs & derivatives , Decorin/physiology , Glioblastoma/pathology , Keratan Sulfate/physiology , Neoplastic Stem Cells/pathology , Neuroblastoma/pathology , Tumor Microenvironment , Dacarbazine/therapeutic use , Humans , Lumican , TemozolomideABSTRACT
Chondroitin sulfate proteoglycans (CSPGs) play important roles in the developing and mature nervous system, where they guide axons, maintain stable connections, restrict synaptic plasticity, and prevent axon regeneration following CNS injury. The chondroitin sulfate glycosaminoglycan (CS GAG) chains that decorate CSPGs are essential for their functions. Through these sugar chains, CSPGs are able to bind and regulate the activity of a diverse range of proteins. CSPGs have been found both to promote and inhibit neuronal growth. They can promote neurite outgrowth by binding to various growth factors such as midkine (MK), pleiotrophin (PTN), brain-derived neurotrophic factor (BDNF) and other neurotrophin family members. CSPGs can also inhibit neuronal growth and limit plasticity by interacting with transmembrane receptors such as protein tyrosine phosphatase σ (PTPσ), leukocyte common antigen-related (LAR) receptor protein tyrosine phosphatase, and the Nogo receptors 1 and 3 (NgR1 and NgR3). These CS-protein interactions depend on specific sulfation patterns within the CS GAG chains, and accordingly, particular CS sulfation motifs are upregulated during development, in the mature nervous system, and in response to CNS injury. Thus, spatiotemporal regulation of CS GAG biosynthesis may provide an important mechanism to control the functions of CSPGs and to modulate intracellular signaling pathways. Here, we will discuss these sulfation-dependent processes and highlight how the CS sugars on CSPGs contribute to neuronal growth, axon guidance, and plasticity in the nervous system.
Subject(s)
Central Nervous System , Chondroitin Sulfate Proteoglycans/physiology , Nerve Regeneration/physiology , Neuronal Plasticity/physiology , Animals , Central Nervous System/cytology , Central Nervous System/growth & development , Central Nervous System/metabolism , Chondroitin Sulfate Proteoglycans/chemistry , HumansABSTRACT
NG2 is purportedly one of the most growth-inhibitory chondroitin sulfate proteoglycans (CSPGs) produced after spinal cord injury. Nonetheless, once the severed axon tips dieback from the lesion core into the penumbra they closely associate with NG2+ cells. We asked if proteoglycans play a role in this tight cell-cell interaction and whether overadhesion upon these cells might participate in regeneration failure in rodents. Studies using varying ratios of CSPGs and adhesion molecules along with chondroitinase ABC, as well as purified adult cord-derived NG2 glia, demonstrate that CSPGs are involved in entrapping neurons. Once dystrophic axons become stabilized upon NG2+ cells, they form synaptic-like connections both in vitro and in vivo. In NG2 knock-out mice, sensory axons in the dorsal columns dieback further than their control counterparts. When axons are double conditioned to enhance their growth potential, some traverse the lesion core and express reduced amounts of synaptic proteins. Our studies suggest that proteoglycan-mediated entrapment upon NG2+ cells is an additional obstacle to CNS axon regeneration.
Subject(s)
Antigens/physiology , Axons/physiology , Cell Communication/physiology , Nerve Regeneration/physiology , Proteoglycans/physiology , Spinal Cord Injuries/physiopathology , Synapses/physiology , Animals , Antigens/genetics , Axons/ultrastructure , Cell Tracking , Cells, Cultured , Chondroitin Sulfate Proteoglycans/physiology , Fibronectins/physiology , Ganglia, Spinal/physiopathology , Ganglia, Spinal/ultrastructure , Integrin beta1/physiology , Laminin/physiology , Mice , Mice, Knockout , Nerve Degeneration/physiopathology , Proteoglycans/geneticsABSTRACT
Mammalian target of rapamycin (mTOR) functions as a master sensor of nutrients and energy, and controls protein translation and cell growth. Deletion of phosphatase and tensin homolog (PTEN) in adult CNS neurons promotes regeneration of injured axons in an mTOR-dependent manner. However, others have demonstrated mTOR-independent axon regeneration in different cell types, raising the question of how broadly mTOR regulates axonal regrowth across different systems. Here we define the role of mTOR in promoting collateral sprouting of spared axons, a key axonal remodeling mechanism by which functions are recovered after CNS injury. Using pharmacological inhibition, we demonstrate that mTOR is dispensable for the robust spontaneous sprouting of corticospinal tract axons seen after pyramidotomy in postnatal mice. In contrast, moderate spontaneous axonal sprouting and induced-sprouting seen under different conditions in young adult mice (i.e., PTEN deletion or degradation of chondroitin proteoglycans; CSPGs) are both reduced upon mTOR inhibition. In addition, to further determine the potency of mTOR in promoting sprouting responses, we coinactivate PTEN and CSPGs, and demonstrate that this combination leads to an additive increase in axonal sprouting compared with single treatments. Our findings reveal a developmental switch in mTOR dependency for inducing axonal sprouting, and indicate that PTEN deletion in adult neurons neither recapitulates the regrowth program of postnatal animals, nor is sufficient to completely overcome an inhibitory environment. Accordingly, exploiting mTOR levels by targeting PTEN combined with CSPG degradation represents a promising strategy to promote extensive axonal plasticity in adult mammals.
Subject(s)
Axons/physiology , Brain Injuries/physiopathology , Nerve Regeneration/physiology , TOR Serine-Threonine Kinases/physiology , Aging/genetics , Aging/physiology , Animals , Brain Injuries/pathology , Chondroitin ABC Lyase/pharmacology , Chondroitin Sulfate Proteoglycans/antagonists & inhibitors , Chondroitin Sulfate Proteoglycans/physiology , Female , Male , Mice , Mice, Knockout , Mice, Transgenic , Nerve Regeneration/drug effects , PTEN Phosphohydrolase/antagonists & inhibitors , PTEN Phosphohydrolase/genetics , PTEN Phosphohydrolase/physiology , Pyramidal Tracts/drug effects , Pyramidal Tracts/injuries , Pyramidal Tracts/physiology , Sirolimus/pharmacology , TOR Serine-Threonine Kinases/antagonists & inhibitorsABSTRACT
Chondroitin sulfate proteoglycans (CSPGs) are a diverse family of extracellular matrix (ECM) molecules that make significant contributions to the patterning and routing of migrating neural cells and extending axons. Three distinct modes of migration mediation result from the relative abundance and positioning of expressed CSPGs, the profile of CSPG receptors expressed by the motile cell types, and the overall way in which the CSPGs integrate into and stabilize the neural ECM. Here we discuss recent findings that help to clarify the molecular mechanisms that underlie these distinct migration-regulating properties as they pertain to neural development, CNS injury, and gliomagenesis.
Subject(s)
Central Nervous System Diseases/metabolism , Central Nervous System , Chondroitin Sulfate Proteoglycans/physiology , Neoplasms/metabolism , Animals , Cell Movement/physiology , Central Nervous System/cytology , Central Nervous System/growth & development , Central Nervous System/metabolism , Humans , Models, Biological , Neurons/metabolism , Neurons/physiologyABSTRACT
BACKGROUND AND OBJECTIVE: Periodontal ligament (PDL) fibroblasts establish principal fibers of the ligament during tooth eruption, and maintain these fibers during occlusion. PDL development and occlusal adaptation includes changes in the orientation of PDL fibroblasts; however, the mechanism for these changes in orientation is unclear. The objective of this study was to compare PDL fibroblast orientation in different stages corresponding with first molar eruption and occlusion in CD44 wild-type (WT) and knockout (KO) mice. MATERIAL AND METHODS: CD44 WT and KO mice were raised to six postnatal stages corresponding with first molar (M1 ) eruption (postnatal day 8, 11, 14 and 18) and occlusion (postnatal day 26 and 41). Coronal sections of the first mandibular molar (M1 ) were prepared and the orientation of fibroblasts in the cervical root region was measured. Angle measurements were compared across developmental stages and between strains using Watson-Williams F-test (oriana software) and ANCOVA. RESULTS: PDL fibroblast orientation increased significantly in CD44 WT (9-87°) and KO mice (14-93°; p ≤ 0.05) between intraosseous eruption (day 11), mucosal penetration (day 14) and preocclusal eruption (day 18); however, the PDL fibroblast orientation did not change significantly with the onset of occlusion (day 26) or continued function (day 41). Within each strain, the variance in fibroblast orientation during preocclusal eruption (day 18) was significantly higher than the variance of all other time points (p < 0.0005). CD44 WT and KO mice showed a similar pattern of PDL development and eruption with a significant difference in CD44 WT vs. KO fibroblast orientations only during early function (day 26, 92° vs 116°; p = 0.05). CONCLUSIONS: The development of PDL fibroblast orientation is highly similar between CD44 WT and KO mice. Between early (day 11) and late (day 18) eruptive stages PDL fibroblast orientation increases, corresponding with the upward movement of M1 . The PDL fibroblast orientation established in preocclusal eruption (day 18) is maintained during early (day 26) and late (day 41) stages of occlusal function, suggesting that PDL cells adapt to mechanical loads in the oral cavity before M1 occlusion.
Subject(s)
Chondroitin Sulfate Proteoglycans/physiology , Fibroblasts/physiology , Periodontal Ligament/cytology , Receptors, Cell Surface/physiology , Tooth Eruption/physiology , Alveolar Process/cytology , Alveolar Process/physiology , Animals , Cell-Matrix Junctions/physiology , Chondroitin Sulfate Proteoglycans/genetics , Dental Occlusion , Extracellular Matrix/physiology , Fibroblasts/cytology , Mice , Mice, Inbred C57BL , Mice, Knockout , Molar/physiology , Receptors, Cell Surface/genetics , Time Factors , Tooth Cervix/cytology , Tooth Cervix/physiology , Tooth Crown/cytology , Tooth Crown/physiology , Tooth Root/cytology , Tooth Root/physiologyABSTRACT
After injury of the central nervous system (CNS) in higher vertebrates, neurons neither grow nor reconnect with their targets because their axons or dendrites cannot regenerate within the injured site. In the CNS, the signal from the environment regulating neurite regeneration is not exclusively generated by one molecular group. This signal is generated by the interaction of various types of molecules such as extracellular matrix proteins, soluble factors and surface membrane molecules; all these elements interact with one another generating the matrix's biological state: the extracellular balance. Proteins in the balanced extracellular matrix, support and promote cellular physiological states, including neuritic regeneration. We have reviewed three types of proteins of the extracellular matrix possessing an inhibitory effect and that are determinant of neuritic regeneration failure in the CNS: chondroitin sulfate proteoglycans, keratan sulfate proteoglycans and tenascin. We also review some of the mechanisms involved in the balance of extracellular proteins such as isomerization, epimerization, sulfation and glycosylation as well as the assemblage of the extracellular matrix, the interaction between the matrix and soluble factors and its proteolytic degradation. In the final section, we have presented some examples of the matrix's role in development and in tumor propagation.