Role Of Heparan Sulfate Proteoglycans Research Articles

Differential role of heparan sulfate proteoglycans on aggregated LDL uptake in human vascular smooth muscle cells and mouse embryonic fibroblasts.

Low density lipoprotein (LDL) receptor-related protein (LRP) binds and internalizes aggregated LDL (agLDL) in human vascular smooth muscle cells (VSMCs). To analyze the contribution of proteoglycans (PGs) to agLDL uptake in human VSMCs, in wild-type mouse embryonic fibroblasts (MEF line), and in LRP-deficient mouse embryonic fibroblasts (PEA13 line). PGs in the medium and cellular and extracellular matrix have been isolated by metabolic radiolabeling with [35S]Na2SO4 and characterized by selective digestion with heparinase I and III (4 U/mL each) and chondroitinase ABC (2 U/mL). To examine the contribution of PGs and LRPs to agLDL internalization, nonexpressing and LRP-expressing cells, treated or not with polysaccharidase, were incubated with agLDL (25, 50, and 100 micro g/mL) for 18 hours. In human VSMCs, agLDL was unable to induce cholesteryl ester (CE) accumulation in antisense LRP-oligodeoxynucleotide-treated cells, and heparan sulfate (HS)-PG depletion leads to a reduction of the CE accumulation. In mouse fibroblasts, PEA13 compared with MEF showed lower, but still considerable, CE accumulation, and HS-PG depletion almost completely inhibited CE accumulation. In MEF, HS-PGs can function alone as receptors that bind and internalize agLDL in the absence of LRP, but in human VSMCs, although HS-PGs facilitate agLDL binding to the cells, LRP is essential for agLDL internalization.

Developmental roles of heparan sulfate proteoglycans in Drosophila.

The formation of complex patterns in multi-cellular organisms is regulated by a number of signaling pathways. In particular, the Wnt and Hedgehog (Hh) pathways have been identified as critical organizers of pattern in many tissues. Although extensive biochemical and genetic studies have elucidated the central components of the signal transduction pathways regulated by these secreted molecules, we still do not understand fully how they organize gradients of gene activities through field of cells. Studies in Drosophila have implicated a role for heparan sulfate proteoglycans (HSPGs) in regulating the signaling activities and distribution of both Wnt and Hh. Here we review these findings and discuss various models by which HSPGs regulate the distributions of Wnt and Hh morphogens.

Heparan sulphate proteoglycans and spinal neurulation in the mouse embryo.

Heparan sulphate proteoglycans have been implicated in the binding and presentation of several growth factors to their receptors, thereby regulating cellular growth and differentiation. To investigate the role of heparan sulphate proteoglycans in mouse spinal neurulation, we administered chlorate, a competitive inhibitor of glycosaminoglycan sulphation, to cultured E8.5 embryos. Treated embryos exhibit accelerated posterior neuropore closure, accompanied by suppression of neuroepithelial bending at the median hinge point and accentuated bending at the paired dorsolateral hinge points of the posterior neuropore. These effects appear specific, as they can be prevented by addition of heparan sulphate to the culture medium, whereas heparitinase-treated heparan sulphate and chondroitin sulphate are ineffective. Both N- and O-sulphate groups appear to be necessary for the action of heparan sulphate. In situ hybridisation analysis demonstrates a normal distribution of sonic hedgehog mRNA in chlorate-treated embryos. By contrast, patched 1 transcripts are abnormally abundant in the notochord, and diminished in the overlying neuroepithelium, suggesting that sonic hedgehog signalling from the notochord may be perturbed by inhibition of heparan sulphation. Together, these results demonstrate a regulatory role for heparan sulphate in mouse spinal neurulation.

The uptake of lipoprotein-borne phylloquinone (vitamin K1) by osteoblasts and osteoblast-like cells: role of heparan sulfate proteoglycans and apolipoprotein E.

Vitamin K is essential for the gamma-carboxylation of Gla-containing bone proteins such as osteocalcin and a suboptimal vitamin K status has been linked to osteoporosis but nothing is known of how the lipoprotein-borne vitamin accesses the bone matrix. We have studied the mechanism of transport of lipoproteins labeled with [3H]-phylloquinone (vitamin K1 [K1]) into osteoblasts using both tumor-derived cell lines and normal osteoblast-rich cell populations. We also investigated the effect of heparin in this model since long-term heparin treatment causes osteopenia and the anticoagulant is known to impair normal lipoprotein metabolism. Heparinase treatment, which removes heparan sulfate proteoglycans (HSPG), reduced uptake of [3H]-K1 from triglyceride-rich lipoproteins (TRL) and low-density lipoproteins (LDL). The effect of heparin in this model was complex depending on cell type, concentration, and time but, overall, the results were consistent with an inhibition of vitamin K uptake by osteoblasts. Anti-apolipoprotein E (apoE) antiserum reduced uptake of TRL-[3H]-K1 by 55 +/- 4% and LDL-[3H]-K1 uptake by 35 +/- 2%. Exogenous apoE4 increased uptake of TRL-[3H]-K1 by 90 +/- 1% compared with 53 +/- 11% for apoE3 and 52 +/- 5% for apoE2. Our findings show that HSPG on the cell surface and apoE in the lipoprotein particles contribute to lipoprotein-K1 uptake by osteoblasts as is known for lipoprotein uptake by hepatocytes. This mechanism is significant in view of the epidemiological association of both undercarboxylation of osteocalcin and the presence of an apo epsilon4 allele with increased fracture risk and reduced bone mineral density (BMD). The inhibition by heparin of lipoprotein-mediated carriage of vitamin K and possibly other lipids to bone may provide a basis for the future understanding of heparin-induced osteoporosis.

The Binding of the Circumsporozoite Protein to Cell Surface Heparan Sulfate Proteoglycans Is Required for PlasmodiumSporozoite Attachment to Target Cells

The major surface protein of malaria sporozoites, the circumsporozoite protein, binds to heparan sulfate proteoglycans on the surface of hepatocytes. It has been proposed that this binding event is responsible for the rapid and specific localization of sporozoites to the liver after their injection into the skin by an infected anopheline mosquito. Previous in vitro studies performed under static conditions have failed to demonstrate a significant role for heparan sulfate proteoglycans during sporozoite invasion of cells. We performed sporozoite attachment and invasion assays under more dynamic conditions and found a dramatic decrease in sporozoite attachment to cells in the presence of heparin. In contrast to its effect on attachment, heparin does not appear to have an effect on sporozoite invasion of cells. When substituted heparins were used as competitive inhibitors of sporozoite attachment, we found that sulfation of the glycosaminoglycan chains at both the N- and O-positions was important for sporozoite adhesion to cells. We conclude that the binding of the circumsporozoite protein to hepatic heparan sulfate proteoglycans is likely to function during sporozoite attachment in the liver and that this adhesion event depends on the sulfated glycosaminoglycan chains of the proteoglycans.

Syndecans: transmembrane modulators of adhesion and matrix assembly.

Syndecans are transmembrane heparan sulfate proteoglycans (HSPGs) that are present on most cell types. HSPGs have been known for some time to regulate a variety of biological processes, ranging from coagulation cascades, growth factor signaling, lipase binding and activity, cell adhesion to ECM and subsequent cytoskeletal organization, to infection of cells with microorganisms. They are complex molecules, with specific core protein to which a variable number of glycosaminoglycan (GAG) chains are attached. Not only the number of chains varies; although syndecans mainly bear heparan sulfate GAGs, some have additional chondroitin/dermatan sulfate chains. Furthermore, heparan sulfate chains can vary in length, epimerization of glucuronic acid to iduronic acid, overall sulfation of the chains, and position of sulfation of the monosaccharides. Early studies relied on structurally nonspecific approaches such as competition with heparin or treatment with heparinases or chlorate to remove GAG chains or prevent their sulfation, respectively. These approaches defined a number of interacting ligands for heparan sulfates including insoluble ECM components and soluble growth factors and cytokines, and they helped to implicate HSPGs in a variety of biological processes (1–5). Cell surface HSPGs, including syndecans, appear to play modulatory roles, such as presenting growth factors to their primary receptors or increasing the infectivity of viruses by interacting with their primary receptors. In addition, these molecules can modify adhesion mediated by the integrins, the major family of receptors for ECM. This last function establishes a parallel between the syndecans (as well as other cell surface HSPGs) and the secreted matricellular proteins discussed elsewhere in this Perspective series. This is an exciting time to be in the field of syndecan research. Recently, there has been a surge of interest in the role of HSPGs and in identifying both the specific sequences of monosaccharides and sulfation patterns involved in ligand binding. With the cloning of various core proteins, it has become possible to study the features of these proteins that affect the ligand-binding ability and subsequent biological responses (reviewed in ref. 5; Table ​Table1).1). Now that tools are available to analyze individual protein cores, and techniques are becoming available to determine the sequence of their GAG chains, it is becoming evident that the protein and carbohydrate components of these molecules each play specific biological roles. Recent advances in “sequencing” of GAG chains present on individual proteoglycans have begun to allow structure to be correlated with function in these complex molecules. For example, a decasaccharide containing 2-O-sulfated iduronic acid and 6-O-sulfated N-acetylglucosamine is needed to potentiate the interaction of FGF-2 with its receptor. In contrast, a less specific sequence of seven to eight N-sulfated disaccharides containing iduronic acid (with or without 2-O-sulfation) serves to bind the higher affinity heparin-binding domain of fibronectin (HepII; ref. 5). This Perspective will concentrate on the syndecan family of transmembrane HSPGs and their roles in adhesion to ECM and subsequent matrix modification. In addition, it will address how syndecans may be pivotal in coordinating growth factor and adhesion signaling mechanisms. For discussion of some recent advances concerning the roles of cell surface proteoglycans in other fundamental biological processes, see refs. 2, 5, 6, and 7 and references therein. Table 1 Known ligands for HSPGs, where sulfation/epimerization patterns have been determined

Heparan sulfate proteoglycans in the nervous system: their diverse roles in neurogenesis, axon guidance, and synaptogenesis

Development of the mammalian nervous system involves generation of neurons from neural stem cells, migration of generated neurons toward genetically determined locations, extension of axons and dendrites, and establishment of neuronal connectivity. Recent progresses revealed diverse role of heparan sulfate proteoglycans in these processes. This article reviews our current knowledge about the functional roles of heparan sulfate proteoglycans in three critical events in mammalian neural development, namely neurogenesis, axon guidance, and synapse development.

Testosterone-Induced Growth of S115 Mouse Mammary Tumor Cells Is Dependent on Heparan Sulfate

The androgen-induced proliferation of S115 mouse mammary tumor cells has been suggested to involve autocrinic fibroblast growth factor signaling. Heparan sulfate proteoglycans are required for fibroblast growth factor signaling, presumably due to their ability to alter binding of fibroblast growth factors to their receptors. We have investigated the role of heparan sulfate proteoglycans in the testosterone-induced proliferation of S115 cells. We demonstrate that when the cells are treated with sodium chlorate, which inhibits the sulfation of endogenous heparan sulfate proteoglycans, cell growth becomes dependent on exogenous heparin. The shortest heparin oligosaccharides supporting cell growth were octasaccharides, whereas dodecasaccharides were almost as effective as native heparin. The N-, 2-O-, and 6-O-sulfate groups of heparin were all required for full testosterone response. Treatment of S115 cells with chlorate or testosterone did not alter the expression of fibroblast growth factor receptors 1 or 3, whereas the expression of fibroblast growth factor receptor 2 was down-regulated. We have previously shown that overexpression of syndecan-1 heparan sulfate proteoglycan renders S115 cells insensitive to testosterone and now demonstrate that this effect can be overcome by sodium chlorate treatment in combination with exogenous heparin. Our results suggest that heparin-like molecules are intimately involved in the androgen-mediated proliferation of S115 cells.

Role of heparan sulphate proteoglycans as potential receptors for non-piliated Pseudomonas aeruginosa adherence to non-polarised airway epithelial cells.

Tight junctions seal polarised surface epithelial respiratory cells so as to prevent the passage of bacteria and toxins through the epithelial sheet. Disruption of tight junctions, which may occur during injury and repair processes of airway epithelium, favours potential bacterial interaction with receptors from cell basolateral membranes. Earlier studies reported that non-polarised and untight epithelial respiratory cells are highly susceptible to Pseudomonas aeruginosa adherence and internalisation. As heparan sulphate proteoglycans (HSP) from cell basolateral membranes in epithelial cells without tight junctions may become accessible to bacterial ligands, the present study investigated their role as potential receptors for non-piliate P. aeruginosa ligands. Treatment of cells with heparitinase I and II significantly reduced (51.2% and 51.7%, respectively) P. aeruginosa adherence to epithelial respiratory cells without tight junctions. The internalisation of bacteria was not affected by treatment with heparitinases. Treatment of the bacteria with heparin and heparan sulphate also significantly reduced their adherence to respiratory cells (34.3% and 43.7%, respectively). Treatment of cells with other enzymes (trypsin, lipase and chondroitinase ABC) or treatment of bacteria with chondroitin-4-sulphate did not modify the adherence to respiratory cells significantly. Both affinity chromatography and Western blotting assays showed the interaction of different P. aeruginosa outer-membrane proteins (OMPs) with heparin. Several bacterial strains showed differences in their profile of heparin-binding OMPs, but all exhibited low mol. wt (< 30 kDa) reactive proteins. Reactivity of whole bacterial cells with heparin was also observed by transmission electron microscopy. These results suggest that HSP are potential receptors for P. aeruginosa adherence to non-polarised and untight epithelial respiratory cells.

Interaction of prion proteins with cell surface receptors, molecular chaperones, and other molecules

This chapter summarizes PrP interacting molecules that are relevant for PrP pathogenesis or transmissible spongiform encephalopathie (TSE) therapy. It describes putative prion protein receptors, including the role of heparan sulfate proteoglycans (HSPGs). A cellular model is presented that describes the possible role of prion receptors and prion proteins, including the recently identified PrP-like protein termed “doppel”. The model emphasizes the possible role of PrP and its receptor regarding PrP internalization as well as signal transduction and physiological function, in particular, the 37 kDa laminin receptor precursor (LRP), an up to now unidentified 66 kDa cell surface protein and cadherins, which are discussed as prion receptors that trigger the entry of PrP into scrapie infectable cells. The chapter describes the role of molecular chaperones, including chemical chaperones that catalyze or hamper the conversion process of PrP C to PrP Sc .

Role of heparan sulfate proteoglycans in cell signaling and cancer

Publisher Summary Heparan sulfate proteoglycans (HSPGs) are composed of a protein core modified on specific serine residues by the addition of heparan sulfate (HS) glucosaminoglycans (GAGs) synthesized in the Golgi. Three distinct types of proteins can serve as the HSPG cores: the transmembrane proteins encoded by the syndecan genes, the glycosylphosphatidylinositol (GPI) membrane-bound glypicans, and the extracellular matrix secreted perlecan proteins. The chapter presents the two major advances made in understanding the role of HSPGs in development and cancer. First, studies in Drosophila have identified many mutations in either the biosynthetic enzymes for example, Sugarless (Sgl), Sulfateless (Sfl), Tout velu (Ttv); or the protein cores for example, syndecans and glypicans. Analysis of the mutant phenotypes has revealed the critical roles of HSPGs in modulating various growth factor signaling pathways. Second, many mutations linked to human cancers have been isolated and shown to correspond to defects in the biosynthesis of HSPGs. The chapter explains the importance of HSPGs in cell signaling and provides insights into their functions. Modulating the activity of HSPGs could influence the development of cancers caused by aberrant cell signaling. The chapter concludes that HSPGs constitute important targets for therapeutics to treat some human tumors.

Cathepsin B Activity Regulation: HEPARIN-LIKE GLYCOSAMINOGLYCANS PROTECT HUMAN CATHEPSIN B FROM ALKALINE pH-INDUCED INACTIVATION

It has been shown that lysosomal cysteine proteinases, specially cathepsin B, has been implicated in a variety of diseases involving tissue remodeling states, such as inflammation, parasite infection, and tumor metastasis, by degradation of extracellular matrix components. Recently, we have shown that heparin and heparan sulfate bind to papain specifically; this interaction induces an increase of its alpha-helix content and stabilizes the enzyme structure even at alkaline pH (Almeida, P. C., Nantes, I. L., Rizzi, C. C. A., Júdice, W. A. S., Chagas, J. R., Juliano, L., Nader, H. B., and Tersariol, I. L. S. (1999) J. Biol. Chem. 274, 30433-30438). In the present work, a combination of circular dichroism analysis, affinity chromatography, cathepsin B mutants, and fluorogenic substrate assays were used to characterize the interaction of human cathepsin B with glycosaminoglycans. The nature of the cathepsin B-glycosaminoglycans interaction was sensitive to the charge and type of polysaccharide. Like papain, heparin and heparan sulfate bind cathepsin B specifically, and this interaction reduces the loss of cathepsin B alpha-helix content at alkaline pH. Our data show that the coupling of cathepsin B with heparin or heparan sulfate can potentiate the endopeptidase activity of the cathepsin B, increasing 5-fold the half-life (t(12)) of the enzyme at alkaline pH. Most of these effects are related to the interaction of heparin and heparan sulfate with His(111) residue of the cathepsin B occluding loop. These results strongly suggest that heparan sulfate may be an important binding site for cathepsin B at cell surface, reporting a novel physiological role for heparan sulfate proteoglycans.

Role of heparan sulfate proteoglycans in cell-cell signaling in Drosophila.

Heparan sulfate proteoglycans (HSPGs) are abundant molecules associated with the cell surface and extracellular matrix, and consist of a protein core to which heparan sulfate (HS) glycosaminoglycan (GAG) chains are attached. Although these molecules have been the focus of intense biochemical studies in vitro, their biological functions in vivo were unclear until recently. We have undertaken an in vivo functional study of HSPGs in Drosophila. Our studies, as well as others, demonstrate the critical roles of HSPGs in several major signaling pathways, including ibroblast growth factor (FGF), Wnt, Hedgehog (Hh) and TGF-beta. Our results also suggest that specific HS GAG chain modifications, as well as specific HSPG protein cores, are involved in specific signaling pathways.

Binding of rat thyroglobulin to heparan sulfate proteoglycans.

We previously showed that rat thyroglobulin (Tg) is a heparin-binding protein and that heparin inhibits Tg binding to megalin (gp330), an endocytic Tg receptor found on the apical surface of thyrocytes. Cooperation between cell surface receptors and heparin-like molecules, namely heparan sulfate proteoglycans (HSPGs), can facilitate cell surface binding of some heparin-binding proteins. Based on our previous findings indicating that heparin and megalin-binding sites of rat Tg are functionally related, here we investigated whether rat Tg binds to HSPGs, which are expressed by thyroid cells. We showed in solid phase assays that unlabeled rat Tg binds to a heparan sulfate (HS) preparation in a dose-dependent, saturable manner, with moderately high affinity (Kd approximately 19 nM, Ki approximately 25 nM). Binding was inhibited by heparin and by HS itself. We then studied the role of HSPGs in Tg binding to FRTL-5 cells, a differentiated Fisher rat thyroid cell line. As previously reported, after incubation of FRTL-5 cells with unlabeled rat Tg at 4 degrees C, heparin released virtually all the cell-bound Tg. Co-incubation of Tg with HS or with a preparation of HSPGs resulted in a reduction of binding by 35%-40%. When FRTL-5 cells were preincubated with heparitinase or heparinase I, which released 20%-30% of cell surface HSPGs, Tg binding was reduced to a similar extent. An antibody against a Tg heparin-binding site functionally related to a major megalin-binding site virtually abolished Tg binding to HS and to FRTL-5 cells, supporting the hypothesis that combined interactions of Tg with HSPGs and with megalin are involved in Tg binding to rat thyroid cells.

Glycosaminoglycans: use in treatment of diabetic nephropathy.

Glycosaminoglycans: use in treatment of diabetic nephropathy.

Role of heparan sulfate proteoglycan on placental thromboxane production

International Journal of Gynecology & ObstetricsVolume 70, Issue S3 p. C24-C24 Free communication Pregnancy - pathophysiology Role of heparan sulfate proteoglycan on placental thromboxane production M. Yamaguchi, M. YamaguchiSearch for more papers by this authorT. Ikenoue, T. IkenoueSearch for more papers by this authorN. Mori, N. MoriSearch for more papers by this author M. Yamaguchi, M. YamaguchiSearch for more papers by this authorT. Ikenoue, T. IkenoueSearch for more papers by this authorN. Mori, N. MoriSearch for more papers by this author First published: 10 December 2003 https://doi.org/10.1016/S0020-7292(00)81481-2AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat No abstract is available for this article. Volume70, IssueS32000Pages C24-C24 RelatedInformation

Cell surface heparan sulfate proteoglycans and lipoprotein metabolism.

Cell surface heparan sulfate proteoglycans are involved in several aspects of the lipoprotein metabolism. Most of the biological activities of these proteoglycans are mediated via interactions of their heparan sulfate moieties with various protein ligands, including lipoproteins and lipases. The binding of lipoproteins to heparan sulfate is largely determined by their apoprotein composition, and apoproteins B and E display the highest affinity for heparan sulfate. Interactions of lipoproteins with heparan sulfate are important for the cellular uptake and turnover of lipoproteins, in part by enhancing the accessibility of lipoproteins to lipoprotein receptors and lipases. Apoprotein B may interact with receptors without involving heparan sulfate. Heparan sulfate has been further implicated in presentation and stabilization of lipoprotein lipase and hepatic lipase on cell surfaces and in the transport of lipoprotein lipase from extravascular cells to the luminal surface of the endothelia. In atherosclerosis, heparan sulfate is intimately involved in several events important to the pathophysiology of the disease. Heparan sulfate thus binds and regulates the activity of growth factors, cytokines, superoxide dismutase and antithrombin, which contribute to aberrant cell proliferation, migration and matrix production, scavenging of reactive oxygen radicals and thrombosis. In this review we discuss the various roles of heparan sulfate proteoglycans in vascular biology, with emphasis on interactions of heparan sulfate with lipoproteins and lipases and the molecular basis of such interactions.

Putting two and two together: crystal structure of the FGF-receptor complex.

The recently determined crystal structure of an FGF-receptor complex reveals a surprising architecture and a novel mode of receptor dimerization. The structure also elucidates the role of heparan sulfate proteoglycans in receptor activation, showing significant differences from previously proposed models.

Heparin-binding EGF-like growth factor interacts with mouse blastocysts independently of ErbB1: a possible role for heparan sulfate proteoglycans and ErbB4 in blastocyst implantation.

Blastocyst implantation requires molecular and cellular interactions between the uterine luminal epithelium and blastocyst trophectoderm. We have previously shown that heparin-binding EGF-like growth factor (HB-EGF) is induced in the mouse luminal epithelium solely at the site of blastocyst apposition at 16:00 hours on day 4 of pregnancy prior to the attachment reaction (22:00-23:00 hours), and that HB-EGF promotes blastocyst growth, zona-hatching and trophoblast outgrowth. To delineate which EGF receptors participate in blastocyst activation, the toxicity of chimeric toxins composed of HB-EGF or TGF-(&agr;) coupled to Pseudomonas exotoxin (PE) were used as measures of receptor expression. TGF-(&agr;) or HB-EGF binds to EGF-receptor (ErbB1), while HB-EGF, in addition, binds to ErbB4. The results indicate that ErbB1 is inefficient in mediating TGF-(&agr;)-PE or HB-EGF-PE toxicity as follows: (i) TGF-(&agr;)-PE was relatively inferior in killing blastocysts, 100-fold less than HB-EGF-PE, (ii) analysis of blastocysts isolated from cross-bred egfr+/- mice demonstrated that HB-EGF-PE, but not TGF-(&agr;)-PE, killed egfr-/- blastocysts, and (iii) blastocysts that survived TGF-(&agr;)-PE were nevertheless killed by HB-EGF-PE. HB-EGF-PE toxicity was partially mediated by cell surface heparan sulfate proteoglycans (HSPG), since a peptide corresponding to the heparin-binding domain of HB-EGF as well as heparitinase treatment protected the blastocysts from the toxic effects of HB-EGF-PE by about 40%. ErbB4 is a candidate for being an HB-EGF-responsive receptor since RT-PCR analysis demonstrated that day 4 mouse blastocysts express two different erbB4 isoforms and immunostaining with anti-ErbB4 antibodies confirmed that ErbB4 protein is expressed at the apical surface of the trophectoderm cells. It is concluded that (i) HB-EGF interacts with the blastocyst cell surface via high-affinity receptors other than ErbB1, (ii) the HB-EGF interaction with high-affinity blastocysts receptors is regulated by heparan sulfate, and (iii) ErbB4 is a candidate for being a high-affinity receptor for HB-EGF on the surface of implantation-competent blastocysts.

Expression patterns of transmembrane and released forms of neuregulin during spinal cord and neuromuscular synapse development

We mapped the distribution of neuregulin and its transmembrane precursor in developing, embryonic chick and mouse spinal cord. Neuregulin mRNA and protein were expressed in motor and sensory neurons shortly after their birth and levels steadily increased during development. Expression of the neuregulin precursor was highest in motor and sensory neuron cell bodies and axons, while soluble, released neuregulin accumulated along early motor and sensory axons, radial glia, spinal axonal tracts and neuroepithelial cells through associations with heparan sulfate proteoglycans. Neuregulin accumulation in the synaptic basal lamina of neuromuscular junctions occurred significantly later, coincident with a reorganization of muscle extracellular matrix resulting in a relative concentration of heparan sulfate proteoglycans at endplates. These results demonstrate an early axonal presence of neuregulin and its transmembrane precursor at developing synapses and a role for heparan sulfate proteoglycans in regulating the temporal and spatial sites of soluble neuregulin accumulation during development.