Extracellular matrix heparin induces alteration of the cell adhesion during brain development (original) (raw)
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Archives of Biochemistry and Biophysics, 2000
The effects of glycosaminoglycans (GAG) on cell-tosubstratum adhesion and neurite elongation were examined in primary cultures of fetal rat hippocampal neurons using tissue culture dishes coated with GAGs coupled to dipalmitoylphosphatidylethanolamine (PE), a novel probe for biological functions of GAGs. Both chondroitin sulfate conjugate to PE (CS-PE) and hyaluronic acid conjugate to PE (HA-PE) promoted neurite elongation from neurons in a dose-dependent manner when immobilized onto polylysine-coated dishes at various concentrations up to 1.0 g/ml. The coating of CS-PE or HA-PE at a concentration higher than 1.0 g/ml resulted in failure of neurite extension and adhesion of neurons to the substrata. In contrast, heparin conjugate to PE (HP-PE) did not exert any effects on neurite elongation or on cell attachment at these concentrations. These findings suggest that GAGs serve as a modulator for neurite elongation during neuronal network formation in the developing central nervous system.
The Journal of cell biology, 1994
We have previously shown that aggregation of microbeads coated with N-CAM and Ng-CAM is inhibited by incubation with soluble neurocan, a chondroitin sulfate proteoglycan of brain, suggesting that neurocan binds to these cell adhesion molecules (Grumet, M., A. Flaccus, and R. U. Margolis. 1993. J. Cell Biol. 120:815). To investigate these interactions more directly, we have tested binding of soluble 125I-neurocan to microwells coated with different glycoproteins. Neurocan bound at high levels to Ng-CAM and N-CAM, but little or no binding was detected to myelin-associated glycoprotein, EGF receptor, fibronectin, laminin, and collagen IV. The binding to Ng-CAM and N-CAM was saturable and in each case Scatchard plots indicated a high affinity binding site with a dissociation constant of approximately 1 nM. Binding was significantly reduced after treatment of neurocan with chondroitinase, and free chondroitin sulfate inhibited binding of neurocan to Ng-CAM and N-CAM. These results indica...
The Journal of biological chemistry, 1994
HB-GAM (heparin binding growth-associated molecule; pleiotrophin) is a secretory, extracellular matrix-associated protein that is strongly expressed in developing nervous tissues and belongs to a novel family of differentiation/growth factors. It promotes axonal growth from perinatal rat brain neurons and is suggested to be mitogenic for some cell types and to display cell-transforming activity. Since the receptors of HB-GAM in cells are unknown, we have started isolation of putative cell surface receptors from brain neurons and from perinatal rat brain. For this purpose, recombinant HB-GAM was produced with the aid of a baculovirus vector and used as an affinity matrix in receptor isolation. A detergent-solubilized component from cultured brain neurons and from brain was identified that binds specifically to HB-GAM and migrates on sodium dodecyl sulfate-polyacrylamide gel electrophoresis as a broad smear with an apparent molecular mass of about 200 kDa. This cell surface component ...
Multiple influences of a heparin-binding growth factor on neuronal development
Journal of Cell Biology, 1987
Heparin-binding growth factor-2 (HBGF-2; Peptides were synthesized as described in Materials and Methods and tested for their ability to block HBGF-2 binding and to stimulate adhesion. To determine the effect of the peptides on HBGF-2 binding, PC12 cells were incubated for 10 min at 4°C in the presence of the indicated synthetic peptides, followed by the addition of ~25I-HBGF-2. The amount of HBGF-2 binding was determined as in Materials and Methods and the data represented as the percent inhibition of binding by the peptides relative to the control cultures. Peptides were used at 100 I.tg/well. To assay the stimulation of cellular adhesion by synthetic peptide fragments of HBGF-2, the indicated peptides were adsorbed to plastic petri dishes at 300 I.tg/35-mm culture dish. All of the input peptides, except J, adhered to the plastic as determined by HPLC or protein analysis. The adhesion of input cells after 1-h incubation was determined as in and presented as the percent stimulation relative to neutral petri dish surfaces where 6-8% of the input cells adhered. All experiments were repeated at least three times with similar results. NS, not significant. A, amphiphilic.
Neuroscience Letters, 1995
Biochemical and cell biological studies have previously identified N-syndecan as a neuronal cell surface receptor in neurite outgrowth induced by heparin-binding growth-associated molecule (HB-GAM). In the present study we have compared temporal and spatial expression patterns of N-syndecan and HB-GAM using Northern and Western blotting, and immunohistochemistry. Expression of Nsyndecan mRNA and protein peaks during the perinatal developmental stage of the brain in the same manner as the expression of HB-GAM mRNA and protein. In addition, both proteins are preferentially localized to fiber tracts of developing brain. We suggest that HB-GAM and N-syndecan form ligand-receptor complexes in developing axon tracts of brain.
The Journal of Cell Biology
Cellular interactions with fibronectin-treated substrata have a complex molecular basis involving multiple domains. A carboxy-terminal cell and heparin binding region of fibronectin (FN) is particularly interesting because it is a strong promoter of neurite outgrowth (Rogers, S. L., J. B. McCarthy, S. L. Palm, L. T. Furcht, and P. C. Letourneau, 1985. J. Neurosci. 5:369-378) and cell attachment (McCarthy, J. B., S. T. Hagen, and L. T. Furcht. 1986. J. Cell Biol. 102:179-188.). To further understand the molecular mechanisms of neuronal interactions with this region of FN, we screened two peptides from the 33-kD heparin binding fragment of the FN A chain, FN-C/H II (KNNQKSEPLIGRKKT) and CS1 (Humphries, M. J., A. Komoriya, S. K. Akiyama, K. Olden, and K. M. Yamada. 1987. J. Biol. Chem. 262:6886-6892), for their ability to promote B104 neuroblastoma cell-substratum adhesion and neurite outgrowth. Both FN-C/H U and CS1 promoted B104 cell attachment in a concentration-dependent and saturable manner, with attachment to FN-C/H II exceeding attachment to CS1. In solution, both exogenous FN-C/H II 1. Abbreviations used in this paper: CNS, central nervous system; ECM, extracellular matrix; FN, fibroneetin; KLH, keyhole limpet hemocyanin; OA, ovalbumin; PNS, peripheral nervous system.
Journal of Neuroscience Research, 1994
Cell-surface heparan sulfate proteoglycans (HSPGs) are potential mediators of neuronal cell adhesion, spreading, and neurite outgrowth on various extracellular matrix molecules. One possible site of HSPG attachment is a heparin binding domain of fibronectin, which is present in the synthetic peptide FN-C/H 11. In this study, HSPGs extracted from embryonic rat spinal cord by detergent were purified by ionexchange chromatography, gel filtration, and affinity chromatography on an agarose column coupled with FN-C/H I1 conjugated to ovalbumin (OA). Heparitinase treatment of the iodinated HSPG fraction led to the appearance of a major protein core with a molecular size of 72 kDa, as determined by reducing SDS-PAGE. The intact proteoglycan has a molecular size of approximately 150-1 65 kDa, containing heparan sulfate glycosaminoglycan chains of about 10-15 kDa. Anti-HSPG antibodies recognized the 72 kDa core protein by immunoblotting, and stained the surface of spinal cord neurons, oligodendrocytes, and a subset of astrocytes. These results identify a cell-surface HSPG that may mediate neuron-substratum or neuron-glia interactions in embryonic central nervous system.
Extracellular adhesive molecules in neurite growth
Bioscience Reports, 1989
This review deals with two topics: (1) the effects of fibronectin and laminin on neurite growth and the molecular mechanisms of these effects, and (2) isolation and properties of the adhesive molecule p30. This novel molecule is an abundant heparin-binding protein in perinatal rat brain, and is suggested to have a role in neuronal growth.
European Journal of Biochemistry, 2001
Polysialylation of the neural cell adhesion molecule (N-CAM) is known to destabilize cell-cell adhesion and to promote plasticity in cell-cell interactions. To gain more insights into the molecular mechanisms regulating the selective expression of polysialic acid on distinct glycan chains, the underlying core structures of polysialylated N-CAM glycans from newborn mouse brain were examined. Starting from low picomolar amounts of oligosaccharides, a multistep approach was used that was based on various mass spectrometric techniques with minimized sample consumption. Evidence could be provided that polysialylated murine N-CAM glycans comprise diantennary, triantennary and tetraantennary core structures carrying, in part, type-1 N-acetyllactosamine antennae, sulfate groups linked to terminal galactose or subterminal N-acetylglucosamine residues and, as a characteristic feature, a sulfated glucuronic acid unit which was bound exclusively to C3 of terminal galactose in Mana3-linked type-2 antennae. Hence, our results reveal that part of the murine N-CAM carbohydrates are modified within a single oligosaccharide by polysialic acid plus a HSO 3-GlcA-moiety, which is likely to represent a HNK1-epitope. As HNK1-carbohydrates are also known to modulate cell-cell interactions, the simultaneous presence of both carbohydrate epitopes may reflect a new mechanism involved in the fine-tuning of N-CAM functions.