Cholinesterases regulation in the absence of ColQ.
Normal physiological activity of the neuromuscular junction (NMJ) requires that key molecules are clustered at the synapse. One of these molecules is acetylcholinesterase (AChE) that regulates acetylcholine levels. This enzyme exists under different isoforms but the predominant form at the NMJ is a collagen-tailed enzyme. The collagen associated to AChE (ColQ) fulfills two functions. It anchors and accumulates AChE in the extracellular matrix. Mutations in ColQ lead to faint or no activity of AChE in the synaptic cleft. As a consequence, normal NMJ functioning is impaired and myasthenic syndromes are observed in patients bearing these mutations. Here, we investigated the effects of ColQ deficiency on cholinesterases mRNA levels and cluster formation. We show that overexpression of AChE but not ColQ in muscle cells is sufficient to drive the formation of AChE clusters. The absence of ColQ in muscle cells in vitro and in vivo leads to an increase in AChE(R) and AChE(T) mRNAs, corresponding to two isoforms of AChE. However, AChE activity is decreased in the medium of ColQ-deficient cells suggesting that AChE secretion is impaired. Butyrylcholinesterase (BChE) mRNAs are also upregulated in vivo. Since AChE and BChE can associate with PRiMA, a membrane anchor, we explored the pattern of expression of PRiMA in vitro and in vivo. The level of PRiMA transcripts is downregulated in the absence of ColQ. Therefore, AChE, BChE and PRiMA mRNA level modifications found in the absence of ColQ cannot compensate for the physiological defects observed at the ColQ-deficient NMJs.
Laboratoire de biologie des jonctions neuromusculaires normales et pathologiques, Université Paris Descartes, INSERM U686, 45, rue des Saints Pères, 75270 Paris, France.
Séverine M
SM
Sigoillot
Francine
F
Bourgeois
Claire
C
Legay
D000110:Acetylcholinesterase / Q000737:chemistry / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D002091:Butyrylcholinesterase / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D003094:Collagen / Q000172:deficiency* / Q000235:genetics; D015536:Down-Regulation; D014644:Genetic Variation; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D009132:Muscles / Q000166:cytology; D009419:Nerve Tissue Proteins / Q000378:metabolism; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D012995:Solubility; D015854:Up-Regulation
D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; C486718:prima1 protein, mouse; D003094:Collagen; D000110:Acetylcholinesterase; C533412:ColQ protein, rat; D002091:Butyrylcholinesterase