Restricted localization of proline-rich membrane anchor (PRiMA) of globular form acetylcholinesterase at the neuromuscular junctions - contribution and expression from motor neurons

• Published in: The FEBS journal 2009-06, Vol.276 (11), p.3031-3042
• Main Authors: Leung, K. Wing, Xie, Heidi Q, Chen, Vicky P, Mok, Mokka K.W, Chu, Glanice K.Y, Choi, Roy C.Y, Tsim, Karl W.K
• Format: Article
• Language: English
• Subjects: acetylcholinesterase; Acetylcholinesterase - chemistry; Acetylcholinesterase - metabolism; Alternative Splicing; Animals; Biochemistry; Blotting, Western; Cell Line; Enzyme-Linked Immunosorbent Assay; Enzymes; Gene Expression Regulation, Developmental; Humans; Membrane Proteins - genetics; Membrane Proteins - metabolism; Membranes; Mice; Microscopy, Confocal; molecular form; Motor Neurons - metabolism; Muscle Denervation; muscle fiber type; Muscle Fibers, Fast-Twitch - metabolism; Muscle Fibers, Slow-Twitch - metabolism; Muscle, Skeletal - growth & development; Muscle, Skeletal - innervation; Muscle, Skeletal - metabolism; Muscular system; Nerve Tissue Proteins - genetics; Nerve Tissue Proteins - metabolism; neuromuscular junction; Neuromuscular Junction - metabolism; proline-rich membrane anchor; Protein Binding; Rats; Rats, Sprague-Dawley; Reverse Transcriptase Polymerase Chain Reaction; Rodents; Spinal Cord - growth & development; Spinal Cord - metabolism
• ISSN: 1742-464X; 1742-4658
• DOI: 10.1111/j.1742-4658.2009.07022.x

The expression and localization of the proline-rich membrane anchor (PRiMA), an anchoring protein of tetrameric globular form acetylcholinesterase (G₄ AChE), were studied at vertebrate neuromuscular junctions. Both muscle and motor neuron contributed to this synaptic expression pattern. During the development of rat muscles, the expression of PRiMA and AChET and the enzymatic activity increased dramatically; however, the proportion of G₄ AChE decreased. G₄ AChE in muscle was recognized specifically by a PRiMA antibody, indicating the association of this enzyme with PRiMA. Using western blot and ELISA, both PRiMA protein and PRiMA-linked G₄ AChE were found to be present in large amounts in fast-twitch muscle (e.g. tibialis), but in relatively low abundance in slow-twitch muscle (e.g. soleus). These results indicate that the expression level of PRiMA-linked G₄ AChE depends on muscle fiber type. In parallel, the expression of PRiMA, AChET and G₄ AChE also increased in the spinal cord during development. Such expression in motor neurons contributed to the synaptic localization of G₄ AChE. After denervation, the expression of PRiMA, AChET and G₄ AChE decreased markedly in the spinal cord, and in fast- and slow-twitch muscles.