Characterization and partial purification of a neuronal factor which increases acetylcholine receptor aggregation on cultured muscle cells

Bauer, H.; Daniels, Mathew P.; Pudimat, P. A.; Jacques, Loretta; Sugiyama, Hiroyuki; Christian, Clifford

doi:10.1016/0006-8993(81)90161-x

Cited by 54 publications

(23 citation statements)

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“…AChR-aggregating molecules have also been extracted from other sources, including rat brain (Podleski et al 1978;Kalcheim et al 1982), chick brain and spinal cord (Jessell et al 1979), conditioned medium from neuronal cell cultures (Christian et al 1978;Schaffner and Daniels 1982), and the electromotor system of Torpedo (Connolly et al 1982). The active components from rat brain and neuronal cellconditioned medium have been reported to be proteins over 50 kD (Podleski et al 1978;Bauer et al 1981), whereas the components from chick brain and Torpedo electromotor system appear to be peptides of less than 1 kD (Connolly et al 1982;Buc-Caron et al 1983). The low-molecular-weight peptides affect AChR number as well.…”

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confidence: 94%

Molecular Components of the Synaptic Basal Lamina That Direct Differentiation of Regenerating Neuromuscular Junctions

Nitkin¹,

Wallace²,

Spira³

et al. 1983

Cold Spring Harbor Symposia on Quantitative Biology

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Results of experiments outlined here provide evidence that components of the myofiber basal lamina sheath direct the formation of active zones in regenerating motor nerve terminals and the development of infoldings and the aggregation of AChRs in the plasma membrane of regenerating myofibers. As a step toward identifying the basal lamina molecules that aggregate AChRs, we are now studying an ECM fraction from the Torpedo electric organ that causes AChRs to aggregate on cultured myotubes. We have solubilized and purified the electric organ AChR-aggregating molecules over 1000-fold. Only nanogram amounts of the most purified extracts are required to cause detectable AChR aggregation. We have also shown that similar activity can be extracted in relatively small amounts from muscle. Antiserum raised against the partially purified electric organ material completely blocked and immunoprecipitated the AChR-aggregating activity in extracts of the electric organ and muscle and bound to components of the basal lamina of frog muscle fibers. Although several polypeptides are present in our most purified extracts, an antiserum against polypeptides in the range of 80 kD completely blocked AChR aggregation by soluble extracts of the electric organ. These findings demonstrate the feasibility of isolating molecules from the synapse-rich electric organ that cause AChR aggregation and comparing them by immunological techniques with those in basal lamina at the neuromuscular junction.

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confidence: 94%

Molecular Components of the Synaptic Basal Lamina That Direct Differentiation of Regenerating Neuromuscular Junctions

Nitkin¹,

Wallace²,

Spira³

et al. 1983

Cold Spring Harbor Symposia on Quantitative Biology

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“…Neuroblastoma X glioma hybrid cell line, NG108-15, can be regarded as one of the best models for neuronal cells. NGlO8-15 cells synthesize and release acetylcholine [I], synthesize an enkephalin-like peptide [2], form functional synapses with skeletal muscle cells in culture [ 1,3] synthesize the promoting factor of acetylcholine receptor aggregation in muscle cells [4], and have many kinds of receptors [5-81. Therefore, it is worthwhile to study membrane properties of NG108-15 cells in order to investigate the mechanisms of the differentiation of neuronal cells.…”

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confidence: 99%

The change in membrane fluidity of neuroblastoma × glioma hybrid cells upon cell differentiation

Kawasaki

Amano

1982

FEBS Letters

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“…Receptor aggregation is evident at chicken junctions, but, during the first 24 hr after contact, newly inserted receptors make up the great majority of those present (7). Several laboratories have characterized factors from neural tissue that alter the number or distribution of AcChoRs on cultured myotubes and that may, therefore, play a role in the accumulation of receptors at nascent synapses (8)(9)(10)(11)(12)(13)(14). Little is known, however, about the molecular mechanisms by which these factors act.…”

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Acetylcholine receptor-inducing factor from chicken brain increases the level of mRNA encoding the receptor alpha subunit.

Harris

Falls

Dill-Devor

et al. 1988

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

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A 42-kDa glycoprotein isolated from chicken brain, referred to as acetylcholine receptor-inducing activity (ARIA), that stimulates the rate of incorporation of acetyicholine receptors into the surface of chicken myotubes may play a role in the nerve-induced accumulation of receptors at developing neuromuscular synapses. Using nuclease-protection assays, we have found that ARIA causes a 2-to 16-fold increase in the level of mRNA encoding the a subunit of the receptor, with little or no change in the levels of y-and 8-subunit messengers. ARIA also increases the amount of a putative nuclear precursor of a-subunit mRNA, consistent with an activation of gene transcription. These results suggest that the concentration of a subunit may limit the rate of biosynthesis of the acetylcholine receptors in chicken myotubes. They also indicate that neuronal factors can regulate the expression of receptor subunit genes in a selective manner. Tetrodotoxin, 8-bromo-cAMP, and forskolin also increase the amount of a-subunit mRNA, with little change in the amount of v-and 6-subunit mRNAs. Unlike ARIA, however, these agents have little effect on the concentration of the a-subunit nuclear precursor.Motor neurons induce the accumulation of acetylcholine receptors (AcChoRs) at developing neuromuscularjunctions (reviewed in refs. 1-3). Two mechanisms have been found to contribute to this phenomenon. Motor neurons may promote the aggregation of receptors that were present on the myocyte surface before nerve-muscle contact, and they may increase the insertion of new receptors in the immediate vicinity of the synapse (4). Receptor aggregation is evident at chicken junctions, but, during the first 24 hr after contact, newly inserted receptors make up the great majority of those present (7). Several laboratories have characterized factors from neural tissue that alter the number or distribution of AcChoRs on cultured myotubes and that may, therefore, play a role in the accumulation of receptors at nascent synapses (8)(9)(10)(11)(12)(13)(14). Little is known, however, about the molecular mechanisms by which these factors act.We have purified a 42-kDa glycoprotein from chicken brain, called ARIA (for AcChoR-inducing activity), that stimulates the rate of insertion of AcChoRs into the membrane of chicken myotubes by as much as 5-fold at picomolar concentrations (15). We were interested in determining which steps of receptor biosynthesis were altered by this factor. Synthesis of the AcChoR is a complex process consisting of transcription of the genes encoding each of the four receptor subunits, translation of the corresponding mRNAs, covalent and conformational modification of the polypeptide chains, assembly of the subunits into a pentameric complex, and transport to the cell surface (reviewed in ref. 16). We report here that the 42-kDa ARIA increases the level of mRNA encoding the a but not the 'y or 8 subunits of the AcChoR. MATERIALS AND METHODSCell Culture. Mononucleated cells were dissociated from pectoral muscles of 11-to 12-day-ol...

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Characterization and partial purification of a neuronal factor which increases acetylcholine receptor aggregation on cultured muscle cells

Cited by 54 publications

References 10 publications

Molecular Components of the Synaptic Basal Lamina That Direct Differentiation of Regenerating Neuromuscular Junctions

Molecular Components of the Synaptic Basal Lamina That Direct Differentiation of Regenerating Neuromuscular Junctions

The change in membrane fluidity of neuroblastoma × glioma hybrid cells upon cell differentiation

Acetylcholine receptor-inducing factor from chicken brain increases the level of mRNA encoding the receptor alpha subunit.

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