Purification of axonin-1, a protein that is secreted from axons during neurogenesis.

Rüegg, Markus A.; Stoeckli, Esther T.; Kühn, Thomas; Heller, Martin; Zuellig, Richard A.; Sonderegger, P.

doi:10.1002/j.1460-2075.1989.tb03348.x

Cited by 79 publications

(89 citation statements)

References 27 publications

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“…The anti-proteoglycan ⌬Di-6S monoclonal antibody 3-B-3 was obtained from Seikagaku. The monoclonal antibody RMO-270 against neurofilament-160 kDa (anti-NF-M) (Invitrogen) and goat polyclonal antibodies against axonin-1 ( Ruegg et al, 1989) were used for axon and growth cone labeling. Lectin staining of tissue sections was done with PNA and goat anti-PNA antibodies (both from Vector Laboratories).…”

Section: Methodsmentioning

confidence: 99%

Versican V0 and V1 Direct the Growth of Peripheral Axons in the Developing Chick Hindlimb

Dutt¹,

Cassoly²,

Dours-Zimmermann³

et al. 2011

J. Neurosci.

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Peanut agglutinin-binding disaccharides and chondroitin sulfate mark transient mesenchymal barriers to advancing motor and sensory axons innervating the hindlimbs during chick development. Here we show that the vast majority of these carbohydrates are at the critical stage and location attached to the versican splice variants V0 and V1. We reveal that the isolated isoforms of this extracellular matrix proteoglycan suppress axon extension at low concentrations and induce growth cone collapse and rapid retraction at higher levels. Moreover, we demonstrate that versican V0 and/or V1, recombinantly expressed in collagen-I gels or ectopically deposited in the hindlimbs of chicken embryos in ovo, cause untimely defasciculation and axon stalling. Consequently, severe disturbances of nerve patterning are observed in the versican-treated embryos. Our experiments emphasize the inhibitory capacity of versicans V0 and V1 in axonal growth and evidence for their function as basic guidance cues during development of the peripheral nervous system.

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Section: Methodsmentioning

confidence: 99%

Versican V0 and V1 Direct the Growth of Peripheral Axons in the Developing Chick Hindlimb

Dutt¹,

Cassoly²,

Dours-Zimmermann³

et al. 2011

J. Neurosci.

Self Cite

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“…The following antibodies were used: ␣Tag1 ) (mAb 4D7) at 1:6; ␣Lh2a/Lh2b (Liem et al, 1997) (L1, rabbit ␣pan (p) Lh2) at 1:1000; ␣Isl1/Isl2 (Tsuchida et al, 1994) (K5, rabbit ␣pIsl) at 1:1000; rabbit ␣Math1 (Helms and Johnson, 1998) at 1:500; rabbit ␣Axonin1 (Ruegg et al, 1989) …”

Section: Immunohistochemistrymentioning

confidence: 99%

BMP type I receptor complexes have distinct activities mediating cell fate and axon guidance decisions

2008

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The finding that morphogens, signalling molecules that specify cell identity, also act as axon guidance molecules has raised the possibility that the mechanisms that establish neural cell fate are also used to assemble neuronal circuits. It remains unresolved, however, how cells differentially transduce the cell fate specification and guidance activities of morphogens. To address this question, we have examined the mechanism by which the Bone morphogenetic proteins (BMPs) guide commissural axons in the developing spinal cord. In contrast to studies that have suggested that morphogens direct axon guidance decisions using noncanonical signal transduction factors, our results indicate that canonical components of the BMP signalling pathway, the type I BMP receptors (BMPRs), are both necessary and sufficient to specify the fate of commissural neurons and guide their axonal projections. However, whereas the induction of cell fate is a shared property of both type I BMPRs, axon guidance is chiefly mediated by only one of the type I BMPRs, BMPRIB. Taken together, these results indicate that the diverse activities of BMP morphogens can be accounted for by the differential use of distinct components of the canonical BMPR complex.

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“…ELISA was essentially done as described by Ruegg et al (1989) with the following modifications: mAb 5B1 (10/~g/ml) was immobilized on microtiter plates followed by incubation of c95 proteins at a concentration between 0.5 and 10 ng/ml. As a secondary antibody, purified rabbit anti-C95A4B, IgG at a concentration of 5.6 ~g/ml was used.…”

Section: Quantification Of Agrinmentioning

confidence: 99%

Acetylcholine receptor-aggregating activity of agrin isoforms and mapping of the active site.

Gesemann

Denzer

Rüegg

1995

The Journal of Cell Biology

222

228

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Abstract. Agrin is a basal lamina protein that induces aggregation of acetylcholine receptors (AChRs) and other molecules at the developing neuromuscular junction. Alternative splicing of chick agrin mRNA at two sites, A and B, gives rise to eight possible isoforms of which five are expressed in vivo. Motor neurons express high levels of isoforms with inserts at sites A and B, muscle cells synthesize isoforms that lack amino acids at the B-site. To obtain further insights into the mechanism of agrin-induced AChR aggregation, we have determined the ECs0 (effective concentration to induce half-maximal AChR clustering) of each agrin isoform and of truncation mutants. On chick myotubes, ECs0 of the COOH-terminal, 95-kD fragment of agrinA4a8 was ,'~35 pM, of agrinA4aZ9 ~110 pM and of agrinA4a, --5 nM. While some AChR clusters were observed with 64 nM of agrinA4aO, no activity was detected for agrinAoa0. Recombinant fulllength chick agrin and a 100-kD fragment of ray agrin showed similar ECso values. A 45-kD, COOH-terminal fragment of agrinA4u retained high activity (ECso ----130 pM) and a 21-kD fragment was still active, but required higher concentrations (ECs0 ---13 nM). Unlike the 45-kD fragment, the 21-kD fragment neither bound to heparin nor did heparin inhibit its capability to induce AChR aggregation. These data show quantitatively that agrinA4nS and agrinA4mg, expressed in motor neurons, are most active, while no activity is detected in agrinAoBo, the dominant isoform synthesized by muscle cells. Furthermore, our results show that a fragment comprising site Bs and the most COOHterminal G-like domain is sufficient for this activity, and that agrin domains required for binding to heparin and those for AChR aggregation are distinct from each other.T HE formation of chemical synapses in the nervous system requires exchange of local signals between preand postsynaptic cells. At the neuromuscular junction (NMJ) ~, muscle cells aggregate several proteins, such as acetylcholine receptors (AChRs), acetylcholinesterase, and heparan sulfate proteoglycans at the site of contact between nerve and muscle cell. This process is, at least in part, initiated by the release of molecules from the motor axon. Several lines of evidence suggest that agrin, a component of synaptic basal lamina, is such a molecule. On cultured myotubes, agrin induces aggregation of several molecules that are concentrated at the NMJ including AChRs (Nitkin et al., 1987;Wallace, 1989). Motor neurons synthesize agrin, transport it to the nerve terminal from where it is released to induce AChR aggregation (Magill

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Purification of axonin-1, a protein that is secreted from axons during neurogenesis.

Cited by 79 publications

References 27 publications

Versican V0 and V1 Direct the Growth of Peripheral Axons in the Developing Chick Hindlimb

Versican V0 and V1 Direct the Growth of Peripheral Axons in the Developing Chick Hindlimb

BMP type I receptor complexes have distinct activities mediating cell fate and axon guidance decisions

Acetylcholine receptor-aggregating activity of agrin isoforms and mapping of the active site.

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