PMID- 25746216
OWN - NLM
STAT- MEDLINE
DCOM- 20160610
LR  - 20151217
IS  - 1095-564X (Electronic)
IS  - 0012-1606 (Linking)
VI  - 408
IP  - 2
DP  - 2015 Dec 15
TI  - Using Xenopus tissue cultures for the study of myasthenia gravis pathogenesis.
PG  - 244-51
LID - S0012-1606(15)00087-1 [pii]
LID - 10.1016/j.ydbio.2015.02.017 [doi]
AB  - Myasthenia gravis (MG), the most common autoimmune disease of neuromuscular 
      junction (NMJ), is heterogeneous in terms of pathophysiology, which is determined 
      by the pathogenic antigen of autoantibodies targeting to synaptic proteins at the 
      NMJs. Currently, patients suspected with MG are routinely screened for the 
      presence of autoantibodies against acetylcholine receptor (AChR) or 
      muscle-specific kinase (MuSK) using a cell-based assay (CBA) that involves the 
      expression of target synaptic membrane protein in heterologous cell lines. 
      However, some autoantibodies may only show reactivity for binding to densely 
      clustered AChR in the physiological conformation, while AChR clustering is known 
      to involve signaling events orchestrated by over a dozen of postsynaptic 
      proteins. To improve the existing serological diagnosis of MG, this study 
      explored the possibility of using the well-established Xenopus primary culture 
      system as a novel CBA for MG. Here, by examining the pathogenic effects of four 
      MG human plasma samples, we found that the samples from both seropositive and 
      seronegative MG patients effectively induced the disassembly of aneural AChR 
      clusters in cultured Xenopus muscle cells, as well as the nerve-induced AChR 
      clusters in the nerve-muscle co-cultures. Importantly, the disassembly of AChR 
      clusters was spatio-temporally correlated to the disappearance of actin 
      depolymerizing factor (ADF)/cofilin, an actin regulator involved in AChR 
      trafficking and clustering. Taken together, this study develops a reliable CBA 
      using Xenopus primary cultures for screening the pathogenicity of human MG plasma 
      samples, and providing a platform for investigating the pathogenic mechanisms 
      underlying the endocytic trafficking and degradation of AChRs at NMJs in MG 
      patients.
CI  - Copyright (c) 2015 Elsevier Inc. All rights reserved.
FAU - Yeo, Hwee Li
AU  - Yeo HL
AD  - Department of Physiology, Yong Loo Lin School of Medicine, National University of 
      Singapore, Singapore 117597, Singapore.
FAU - Lim, Jorain Yu Ni
AU  - Lim JY
AD  - Department of Physiology, Yong Loo Lin School of Medicine, National University of 
      Singapore, Singapore 117597, Singapore.
FAU - Fukami, Yuki
AU  - Fukami Y
AD  - Department of Medicine, Yong Loo Lin School of Medicine, National University of 
      Singapore, Singapore 117599, Singapore.
FAU - Yuki, Nobuhiro
AU  - Yuki N
AD  - Department of Physiology, Yong Loo Lin School of Medicine, National University of 
      Singapore, Singapore 117597, Singapore; Department of Medicine, Yong Loo Lin 
      School of Medicine, National University of Singapore, Singapore 117599, 
      Singapore.
FAU - Lee, Chi Wai
AU  - Lee CW
AD  - Department of Physiology, Yong Loo Lin School of Medicine, National University of 
      Singapore, Singapore 117597, Singapore. Electronic address: 
      leechiwai@nuhs.edu.sg.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20150303
PL  - United States
TA  - Dev Biol
JT  - Developmental biology
JID - 0372762
RN  - 0 (Actin Depolymerizing Factors)
RN  - 0 (Autoantibodies)
RN  - 0 (Receptors, Cholinergic)
RN  - EC 2.7.10.1 (MUSK protein, human)
RN  - EC 2.7.10.1 (Receptor Protein-Tyrosine Kinases)
SB  - IM
MH  - Actin Depolymerizing Factors/metabolism
MH  - Animals
MH  - Animals, Genetically Modified
MH  - Autoantibodies/blood
MH  - Humans
MH  - Myasthenia Gravis/*etiology/immunology/metabolism
MH  - Neuromuscular Junction/immunology/metabolism
MH  - Receptor Protein-Tyrosine Kinases/immunology/metabolism
MH  - Receptors, Cholinergic/immunology/metabolism
MH  - Tissue Culture Techniques
MH  - Xenopus/embryology/genetics
OTO - NOTNLM
OT  - ADF/cofilin
OT  - Acetylcholine receptor
OT  - Cell-based assay
OT  - Myasthenia gravis
OT  - Neuromuscular junction
OT  - Xenopus
EDAT- 2015/03/10 06:00
MHDA- 2016/06/11 06:00
CRDT- 2015/03/10 06:00
PHST- 2014/10/31 00:00 [received]
PHST- 2015/02/10 00:00 [revised]
PHST- 2015/02/20 00:00 [accepted]
PHST- 2015/03/10 06:00 [entrez]
PHST- 2015/03/10 06:00 [pubmed]
PHST- 2016/06/11 06:00 [medline]
AID - S0012-1606(15)00087-1 [pii]
AID - 10.1016/j.ydbio.2015.02.017 [doi]
PST - ppublish
SO  - Dev Biol. 2015 Dec 15;408(2):244-51. doi: 10.1016/j.ydbio.2015.02.017. Epub 2015 
      Mar 3.