Simultaneous demonstration of 3 synaptic markers: nerve endings, localization of AChE and AChR clusters at neuromuscular junctions of the rat in vitro

Abstract

Our triple labelling method allows the AChE and AChR distributions and the nerve endings to be stained in the same preparation. This method which provides a detailed visualization of the synaptic contacts and is capable of revealing a weak AChE activity is particularly valuable in studies of synaptogenesis.

Effects of electroacupuncture on recovery of the electrophysiological properties of the rabbit gastrocnemius after contusion: an in vivo animal study

“…Newly formed (i.e., recovering) NMJs have been shown to have the capacity for endplate potentials; however, contraction of myofibers is not achieved [ 28 ], that is, the NMJ did not perform its original function, despite the completion of nerve regeneration and sufficient contact between myofibers and neurons. Moreover, original reconstructed NMJ has been shown to exhibit incomplete function, and the reconstruction of NMJ function (i.e., the impulse) still requires the auxiliary function of nerve and muscle signaling molecules [ 29 - 32 ].…”

“…Thus, it is possible that treatment with EA promoted the recovery of the electrophysiological properties of the GM in addition to enhancing nerve repair and myofiber regeneration. Recovery of NMJ function can also be achieved by increased synthesis of Ach, AchE, and AchR, which accumulate steadily at the postsynaptic membrane [ 32 ].…”

“…Its physiological function is to quickly hydrolyze the neurotransmitter Ach, ultimately causing Ach to activate cholinergic receptors while maintaining the sensitivity of nerve impulses. Therefore, the presence of AchE at the postsynaptic membrane reflects the recovery of NMJ function [ 32 ]. In our study, we examined AchE as a tissue marker to verify the role of EA in facilitating accumulation of AchE at the postsynaptic membrane.…”

“…Agrin also plays a key role in the physiological processes of the NMJ, especially the postsynaptic membrane, which is involved in the formation, differentiation, maturation, and consistency of the NMJ [ 34 ]. The Agrin/muscle-specific receptor tyrosine kinase (MuSK)/Rapsyn signaling pathway has been shown to be critical for postsynaptic gathering and decomposition of the AchR [ 32 ]. Within this pathway, Agrin activates postsynaptic MuSK by interacting with Lrp4 receptors.…”

In Vitro Innervation as an Experimental Model to Study the Expression and Functions of Acetylcholinesterase and Agrin in Human Skeletal Muscle

“…Evidence supporting neuronal origin of AChE in the NMJ was also obtained in co-cultures of embryonic rat spinal cord and human skeletal muscle cells, which provided an alternative approach to tackle the question of the cellular origin of AChE in the NMJ. Unlike some other experimental approaches, in which neuronal or muscular component was removed [ 170 , 186 ] or NMJs were more immature [ 195 ], the rat-human co-cultures contain well-differentiated NMJs comprising the presynaptic and postsynaptic components of rat and human origin, respectively [ 24 ]. By using species-specific antibodies, we were able to distinguish between muscular (i.e., human) and neuronal (i.e., rat) AChE.…”