Effect of Nerve Growth Factor on the Expression of Colchicine-Binding Activity and 14-3-2 Protein in an Established Line of Human Neuroblastoma

Kolber, Alan R.; Goldstein, Milton N.; Moore, Blake W.

doi:10.1073/pnas.71.10.4203

Cited by 43 publications

(10 citation statements)

References 17 publications

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“…The latter, however, show a higher density of microtubules than the undifferentiated cells, indicating that during neurite outgrowth new microtubules are assembled from a pool of preformed tubulin subunits (33). In contrast, the increase in the microtubule population in growing neurites of both human neuroblastoma (39) and chick embryo dorsal root ganglia cultures (41) after treatment with nerve growth factor (NGF) appeared to be correlated with increased synthesis of tubulin subunits, measured by increased colchicine-binding activity. Stimulation of neurite outgrowth by NGF does not occur in the presence of inhibitors of protein synthesis.…”

Section: Discussionmentioning

confidence: 82%

Enhanced labeling of a retinal protein during regeneration of optic nerve in goldfish.

Heacock¹,

Agranoff²

1976

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

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Goldfish retinas were examined for changes in the labeling pattern of protein during regeneration of retinal ganglion cell axons following unilateral optic nerve crush. At various times after optic nerve crush the normal retinas were incubated in vitro with [3H] Following intra-orbital crush of the optic nerve, in goldfish, the retinal ganglion cell must grow out a new axon several millimeters long. This process not only implies a substantial increase in the rate of protein synthesis but may also involve a selective increase in the synthesis of the proteins necessary for the renewal of axoplasm and replacement of structural elements of the axon. Formation of cell-specific markers involved in retinotectal recognition may be increased~during this period (10). There might, in addition, be an initial decrease in the synthesis of components necessary to maintain the functional integrity of the nerve ending until the growing nerve reaches the optic tectum (11).This report presents comparisons of the in vitro protein labeling pattern of normal goldfish retina with that of retinas removed at various times after optic nerve crush (postcrush retina). A double-labeling technique coupled with sodium dodecyl sulfate-urea polyacrylamide gel electrophoresis (12) revealed an increased amino-acid incorporation into a protein fraction with properties of the microtubule subunit, tubulin. MATERIALS AND METHODSGoldfish (Carassius auratus), 6-7 cm in body length, were anesthetized with tricaine methane sulfonate prior to an intraorbital crush of the right optic nerve. The left optic nerve remained intact so that the left retina served as a control. Following surgery, the goldfish were stored in groups of 50 to 80 in 30 gallon tanks at 20-22°, and were fed daily.Fish were dark-adapted for 30 min prior to removal of retinas in order to facilitate separation of the retina from the pigment epithelium. After hemisection of the eye, the retina was floated free from the pigment layer with 0.86% saline and detached by cutting at the optic disk. The retinas were then rinsed and stored in ice-cold saline or incubation medium for up to 1 hr prior to incubation.In were evaporated to dryness under nitrogen immediately before use. After evaporation, each was resuspended in a small volume of incubation medium. The purity of each isotopic preparation was checked by paper chromatography. The incubation medium was that of Dunlop et al. (13): N-2-bydroxyethylpiperazine-N'-2-ethanesulfonic acid (Hepes), 25 mM; MgSO4, 1.3 mM; CaCl2, 2.6 mM; K2HPO4, 1.2 mM; KCI, 5.9 mM; NaCl, 106.5 mM; glucose, 12 mM; and NaOH to pH 7.4.Retinas (in groups of 5 to 20) were pre-incubated for 2 min at 25°in 1.9 ml of incubation medium. Labeled precursor (0.1 ml) was then added (1.0-4.0 ,uCi per retina) and incubation was continued in air in a Dubnoff metabolic shaker at 250. All incubations using double-labeled methionine were carried out for 1 hr. The reaction was stopped by dilution with 2 volumes of ice-cold incubation medium containing 2 mM L-methionine. After c...

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Section: Discussionmentioning

confidence: 82%

Enhanced labeling of a retinal protein during regeneration of optic nerve in goldfish.

Heacock¹,

Agranoff²

1976

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

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“…This enzyme belongs to a new class of surface proteins that do not possess classical machinery for surface transport, but are transported on the cell surface by an unknown mechanism 15 . Enolase has been recently reported to be a multifunctional protein, having such important roles as a neurotrophic factor 16 , a hypoxic stress protein, and a strong plasminogen binding protein 17 . It has also been identified as one of the most frequently identified differentially expressed brain proteins in human and animal tissues 18 .…”

Section: Resultsmentioning

confidence: 99%

Quantitative proteomic analysis of differentially expressed proteins in AÎ²(17-42) treated synaptosomes

et al. 2012

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“…Reference Kolber et al (1974) Brodeur and Goldstein (1976) Goldstein and Brodeur (1974) Perez-Polo et al (1979) Haskell et al (1983) Schulze and Perez-Polo (1982) Reynolds and Perez-Polo (1981) Prasad and Kumar (1975) Goldstein and Brodeur (1974) Goldstein and Brodeur (1974) Prasad and Kumar (1975) Prasad and Kumar (1975) Prasad and Kumar (1975) Prasad and Kumar (1975) Prasad and Kumar (1975) Miyoke et al (1975) Prasad and Kumar (1975) Prasad and Kumar (1975) The purpose of assay systems is to provide an efficient screening paradigm for the copurification of NTFs and neuronotrophic activity prior to further characterization of the biological effects of an NTF. The advantages and disadvantages of tissue explants vs. dissociated cell cultures have been extensively described elsewhere (Barde et al, 1983).…”

Section: Imr-32mentioning

confidence: 99%

Cell Culture in the Neurosciences

Bottenstein¹,

Sato²

1985

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Effect of Nerve Growth Factor on the Expression of Colchicine-Binding Activity and 14-3-2 Protein in an Established Line of Human Neuroblastoma

Abstract: Purified nerve growth factor induced the outgrowth of neurites from cultured human neuroblastoma cells (NJB line) and a concomitant increase in colchicine-binding activity in extracts from these cultures.

Cited by 43 publications

References 17 publications

Enhanced labeling of a retinal protein during regeneration of optic nerve in goldfish.

Enhanced labeling of a retinal protein during regeneration of optic nerve in goldfish.

Quantitative proteomic analysis of differentially expressed proteins in AÎ²(17-42) treated synaptosomes

Cell Culture in the Neurosciences

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