Attachment to an endogenous laminin-like protein initiates sprouting by leech neurons.

Chiquet, Matthias; Masuda-Nakagawa, Liria M.; Beck, Konrad

doi:10.1083/jcb.107.3.1189

Cited by 71 publications

(57 citation statements)

References 36 publications

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“…We prepared an aqueous protein extract of the extracellular matrix of Hirudo medicinalis (20) and spread it on a coverslip. The dry protein was illuminated by a mercury high-pressure lamp through a mask of aluminum stripes (width 10 Am) on silica (P.F.…”

Section: Methodsmentioning

confidence: 99%

Cable properties of a straight neurite of a leech neuron probed by a voltage-sensitive dye.

Fromherz

Müller²

1994

Proc. Natl. Acad. Sci. U.S.A.

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We measured a time-resolved map of electrical activity in a thin straight neurite (1. (24). The 100 outputs were fed into current-voltage converters (OPA 121; Burr-Brown, Filderstadt, F.R.G., with 100-Ma feedback resistors, time constant 0.4 ms) and eventually into differential sample-and-hold amplifiers. Ten diodes were selected, and their signals were stored on a tape recorder (Racal, Bergisch Gladbach, F.R.G.).The protocol of a measurement was as follows: (i) Adjustment of a segment of the neurite along a row of 10 diodes, (ii) record baselines in the dark, (iii) record fluorescence with open shutter (after a 5-ms interval) and then switch to the sample-and-hold mode with the total fluorescence as a reference. (iv) Record and store fluorescence with electrical stimulation for 70 ms: A positive current (2-4 nA) was injected for 25 ms to elicit an action potential (amplitude 60-75 mV). After an interval of 10 ms, a negative Gaussian voltage (amplitude around -100 to -160 mV) was induced by a negative and positive current pulse. (v) After an interval of 5 s the fluorescence without stimulation was recorded for 70 ms and stored (baseline of bleaching). To overcome the limited range of the diode array (-85 pum) we displaced the neuron along a row of diodes several times-starting with the terminal segment-and repeated cycles (i-v). A complete set of data was sampled from the tape at an interval of0.1 ms and a resolution of 12 bit and read into a microcomputer. The fluorescence changes without stimulation were fitted by an exponential function and subtracted from the changes with stimulation; the results were divided by the total fluorescences. The transients of depolarization were corrected for the decreased amplitude of the action potential (from 75 to 60 mV) during a set of measurements with displaced neurites.Then all transients of depolarization were normalized to constant amplitude. The transients of hyperpolarization were scaled by the same factors. Finally the signals were smoothed Abbreviation: N cell, nociceptive neuron. 4604The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

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Section: Methodsmentioning

confidence: 99%

Cable properties of a straight neurite of a leech neuron probed by a voltage-sensitive dye.

Fromherz

Müller²

1994

Proc. Natl. Acad. Sci. U.S.A.

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“…After one month, the mice were boosted twice within two days with the same preparation (in NaCl/P, alone) which was injected into the tail vein. Fusion of spleen cells with PA-1 myeloma cells was carried out as described by Chiquet et al (1988). Hybridoma supernants were screened by ELISA (Chiquet et al, 1988) with DEAE-Sephadex-purified immunogen at 2 -4 pg/ml.…”

Section: Preparation Of Antibodiesmentioning

confidence: 99%

“…Fusion of spleen cells with PA-1 myeloma cells was carried out as described by Chiquet et al (1988). Hybridoma supernants were screened by ELISA (Chiquet et al, 1988) with DEAE-Sephadex-purified immunogen at 2 -4 pg/ml. About 70 positive hybridoma lines were further screened by immunoblotting.…”

Section: Preparation Of Antibodiesmentioning

confidence: 99%

“…Two-dimensional gels (non-reduced against reduced) were carried out as described by Chiquet et al (1988). After electrophoresis, the gels were either soaked in 1 M sodium salycilate and processed for fluorography (Chiquet and Fambrough, 1984b), or transferred to nitrocellulose for immunoblotting (Chiquet et al, 1988). …”

Section: Sds/page and Immunoblottingmentioning

confidence: 99%

“…Specimes were rotary shadowed (Shotton et al 1979) and analyzed using a Zeiss 109 electron microscope as described (Chiquet et al, 1988). Negative staining was performed as follows: Collodium-carbon grids were glow-discharged and wet with buffer (10 mM NaN3, 1 mM MgCl,, 10 mM Tris/ HCl, pH 7.0) for 45 s. Fluid was blotted off with a filter paper and grids were placed on top of a drop of Tris/NaCl solution containing the protein.…”

Section: Electron Microscopymentioning

confidence: 99%

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A major oligomeric fibroblast proteoglycan identified as a novel large form of type‐XII collagen

Koch

Bernasconi

Chiquet

1992

European Journal of Biochemistry

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Cultured chick embryo skin fibroblasts release a major component with a native molecular mass of about 1 MDa, which resolves into three polypeptide bands of about 300, 350 and 600 kDa upon reduction. We report here the purification of this oligomeric protein and show, by means of polyclonal and monoclonal antibodies, that its three polypeptide constituents are closely related. The 600-kDa polypeptide is likely to be a dimer of two smaller subunits which are cross-linked by non-reducible bonds. By electron microscopy, isolated oligomeric molecules exhibit a novel cruciform structure with a large central globular domain. One arm has the shape of a thin rod about 70 nm in length. The three other arms are thicker, longer (90 nm) and flexible, and carry a prominent double globule at their distal ends. Collagenase treatment of the oligomeric fibroblast protein yields two resistant fragments of about 270 kDa and 320 kDa. The intact 350-kDa and 600-kDa (but not the 300-kDa) polypeptides are chondroitinase sensitive and labeled by metabolic incorporation of [35S]sulfate; collagenase treatment does not remove any [35S]sulfate. Hence, the intact fibroblast protein has glycosaminoglycan chains attached to its non-collagenous domain. Three amino acid sequences obtained from chymotryptic fragments of the fibroblast protein correspond to sequences predicted for chick type-XI1 collagen from its full-length cDNA [Yamagata, M., Yamada, K. M., Yamada, S. S., Shinomura, T., Tanaka, H., Nishida, Y., Obara, M. & Kimata, K. (1991) J. Cell Biol. 115, 209-2211. However, the novel fibroblast protein described here differs significantly from previously isolated forms of type-XI1 collagen: its subunits are larger by one third, and it is a proteoglycan.FACIT collagens (fibril-associated collagens with interrupted triple helix) belong to a recently discovered class of extracellular matrix molecules. (Gordon et al., 1987;Dublet and van der Rest, 1987; for review, see Shaw and Olson, 1991). Its members (collagens type IX, XI1 and XIV) are thought to bind to the surface of fibrillar collagen bundles via short collagenous domains, while their large N-terminal non-collagenous parts project outwards to interact with other matrix components or cell surfaces (Vaughan et al., 1988;Keene et al., 1991). The FACIT prototype, type-IX collagen, is a heterotrimeric (Huber et al., 1986) molecule carrying a large non-collagenous glycoprotein domain (Vasios et al., 1988) as well as a chondroitin sulfate chain (Vaughan et al., 1985;Huber et al., 1986), and it is attached to the surface of type-I1 collagen fibrils in cartilage (Vaughan et al., 1988). In type-XI1 collagen purified from chick tendon, the very large Nterminal, non-collagenous domains form a structure with three arms, each 60 nm, which accounts for more than 80% of the mass of the molecule . Recent cDNA sequencing has shown that the large non-collagenous extensions of type-XI1 collagen contain domains homologous

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Outgrowth patterns and directed growth of identified neurons induced by native substrates in culture

Fernández-de-Miguel

1997

J. Comp. Neurol.

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In this study, we have tested how various identified leech neurons in culture grow on surfaces that they normally contact in situ. Neurons were cultured either on ganglion capsules from which neurons had been removed or on skin. On these substrates, outgrowth patterns were characteristic for each cell type. Retzius cells plated on capsules extended bundles of thick, fasciculated processes with few branching points and in the opposite direction a tangle of fine neurites. Anterior pagoda (AP) neurons plated on capsules extended two single processes in opposite directions but failed to grow on skin. Sensory P and N neurons on capsules extended multiple processes. On skin, P neurons extended only two long branches in opposite directions over the superficial body wall. N neurons on skin extended multiple processes. Varicosities were common in the processes of P and N neurons on capsules or skin. The branching patterns described here bore closer resemblance to those in the developing or adult nervous system than to those on Concanavalin A or laminin-enriched extract. Pairs of Retzius or AP neurons plated at a distance on the same capsule extended neurites from one neuron toward the other and formed contacts. Such directed growth failed in hybrid pairs of Retzius and AP neurons or in pairs plated on laminin-enriched extract or Concanavalin A. Our results suggest that multiple growth-promoting molecules anchored to the extracellular matrix may cooperate in regulating the branching pattern of neurons, fasciculation, and direction of growth.

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Attachment to an endogenous laminin-like protein initiates sprouting by leech neurons.

Cited by 71 publications

References 36 publications

Cable properties of a straight neurite of a leech neuron probed by a voltage-sensitive dye.

Cable properties of a straight neurite of a leech neuron probed by a voltage-sensitive dye.

A major oligomeric fibroblast proteoglycan identified as a novel large form of type‐XII collagen

Outgrowth patterns and directed growth of identified neurons induced by native substrates in culture

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