Molecular distinction between fetal and adult forms of muscle acetylcholine receptor

Mishina, Masayoshi; Takai, Toshiyuki; Imoto, Keiji; Noda, Masaharu; Takahashi, Tomoyuki; Numa, Shosaku; Methfessel, Christoph; Sakmann, Bert

doi:10.1038/321406a0

Cited by 942 publications

(498 citation statements)

References 41 publications

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“…Samples of RNA (1Opg each) were denatured in 1 M glyoxal and 50% dimethyl sulfoxide [lo], electrophoresed on 1.5% agarose gels and transferred+ [l l] to Biodyne nylon membranes (Pall). Hybridization was carried out according to Thomas [11] and as detailed by Mishina et al [2]. All hybridization probes were labelled with [cu-32P]dCTP using the random primed labelling kit from Boehringer (Mannheim) which followed labelling procedures based on the method of Feinberg and Vogelstein [12].…”

Section: Rna Isolation and Rna Blot Analysismentioning

confidence: 99%

“…Both subtypes are comprised of the CP, ,& and J-subunits and either the y-or the f-subunit [2-41. In bovine muscle the levels of the mRNA encoding the y-and the csubunit change during development and correlate with the presence of the two AChR subtypes in fetal and adult muscle, respectively [2]. A number of important issues remain unresolved, including…”

Section: Introductionmentioning

confidence: 99%

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Developmental regulation of five subunit specific mRNAs encoding acetylcholine receptor subtypes in rat muscle

et al. 1989

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The muscular content of the mRNAs encoding the five subunits of the nicotinic acetylcholine receptor was measured during postnatal development in the rat. Subunit specific mRNAs show differchtial regulation. The levels of the a-, yand &subunit specific mRNAs decrease steadily after birth, while the & and a-subunit mRNAs increase transiently and then decrease. The adult pattern of subunit specific mRNA levels is reached at 46 weeks postnatally. The content of y-and e-subunit mRNA changes in a reciprocal fashion during the first 2 postnatal weeks, supporting the view that differential regulation of y-and s-subunit mRNA during development is one mechanism mediating the appearance of the adult, s-subunit containing, subtype of end-plate channel. Denervation of neonatal muscle increases the levels of all subunitspecific mRNAs during further development. It prevents the postnatal decrease in y-subunit mRNA and enhances the initial increase in a-subunit mRNA. This makes it appear that the e-subunit gene is less sensitive to regulation by the nerve in the postnatal period than the y-subunit gene.

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Section: Rna Isolation and Rna Blot Analysismentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Developmental regulation of five subunit specific mRNAs encoding acetylcholine receptor subtypes in rat muscle

et al. 1989

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“…The autoantigen involved in MG is the nicotinic acetylcholine receptor (AChR), which is a transmembrane glycoprotein with five homologous subunits in the stoichiometry a2bcd or a2bed [2]. The anti-AChR antibodies in MG cause accelerated internalization and degradation of the AChR (antigenic modulation), complement-mediated destruction of the postsynaptic membrane and blockade of acetylcholine binding to the AChR [3].…”

Section: Introductionmentioning

confidence: 99%

Isolation and characterization of human anti-acetylcholine receptor monoclonal antibodies from transgenic mice expressing human immunoglobulin loci

Protopapadakis¹,

Kokla²,

Tzartos³

et al. 2005

Eur. J. Immunol.

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The isolation of human antibodies against muscle acetylcholine receptor (AChR), the autoantigen involved in myasthenia gravis (MG), is important for the development of therapeutically useful reagents. Monovalent antibody fragments from monoclonal antibodies against the main immunogenic region (MIR) of AChR protect the receptor from the destructive activity of MG autoantibodies. Human anti-AChR a-subunit antibody fragments with therapeutic potential have been isolated using phage display antibody libraries. An alternative approach for obtaining human mAb has been provided by the development of humanized mice. In this report, we show that immunization of transgenic mouse strains with the extracellular domain of the human AChR a-subunit results in antibody responses and isolation of hybridomas producing human mAb. Four specific IgM mAb were isolated and analyzed. mAb170 recognized the native receptor the best and was capable of inducing AChR antigenic modulation, suggesting its specificity for a pathogenic epitope. Moreover, the recombinant antigen-binding (Fab) fragment of this mAb competed with an anti-MIR mAb, revealing that its antigenic determinant lies in or near the MIR. Finally, Fab170 was able to compete with MG autoantibodies and protect the AChR against antigenic modulation induced by MG sera. This approach will be useful for isolating additional mAb with therapeutic potential against the other AChR subunits.

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“…For example, early studies with muscle-type nAChRs containing either the γ or ε subunits helped to explain differences between the ion channel properties of receptors found in embryonic and adult muscle [84]. Similar approaches with neuronal nAChRs have helped to demonstrate that both α and non-α subunits can influence ligand-binding and ion channel properties [144][145][146].…”

Section: The Influence Of Subunit Compositionmentioning

confidence: 99%

A review of experimental techniques used for the heterologous expression of nicotinic acetylcholine receptors

Millar

2009

Biochemical Pharmacology

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Nicotinic acetylcholine receptors (nAChRs) are members of the Cys-loop family of neurotransmitter-gated ion channels, a family that also includes receptors for γ-aminobutyric acid, glycine and 5-hydroxytryptamine. In humans, nAChRs have been implicated in several neurological and psychiatric disorders and are major targets for pharmaceutical drug discovery. In addition, nAChRs are important targets for neuroactive pesticides in insects and in other invertebrates. Historically, nAChRs have been one of the most intensively studied families of neurotransmitter receptors. They were the first neurotransmitter receptors to be biochemically purified and the first to be characterized by molecular cloning and heterologous expression. Although much has been learnt from studies of native nAChRs, the expression of recombinant nAChRs has provided dramatic advances in the characterization of these important receptors. This review will provide a brief history of the characterization of nAChRs by heterologous expression. It will focus, in particular, upon studies of recombinant nAChRs, work that has been conducted by many hundreds of scientists during a period of almost 30 years since the molecular cloning of nAChR subunits in the early 1980's.

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Molecular distinction between fetal and adult forms of muscle acetylcholine receptor

Cited by 942 publications

References 41 publications

Developmental regulation of five subunit specific mRNAs encoding acetylcholine receptor subtypes in rat muscle

Developmental regulation of five subunit specific mRNAs encoding acetylcholine receptor subtypes in rat muscle

Isolation and characterization of human anti-acetylcholine receptor monoclonal antibodies from transgenic mice expressing human immunoglobulin loci

A review of experimental techniques used for the heterologous expression of nicotinic acetylcholine receptors

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