Quantal Secretion and Nerve-Terminal Cable Properties at Neuromuscular Junctions in an Amphibian (Bufo marinus)

1999 ◽  
Vol 81 (3) ◽  
pp. 1135-1146 ◽  
Author(s):  
G. T. Macleod ◽  
L. Farnell ◽  
W. G. Gibson ◽  
M. R. Bennett

Quantal secretion and nerve-terminal cable properties at neuromuscular junctions in an amphibian ( Bufo marinus). The effect of a conditioning depolarizing current pulse (80–200 μs) on quantal secretion evoked by a similar test pulse at another site was examined in visualized motor-nerve terminal branches of amphibian endplates ( Bufo marinus). Tetrodotoxin (200 nM) and cadmium (50 μM) were used to block voltage-dependent sodium and calcium conductances. Quantal release at the test electrode was depressed at different distances (28–135 μm) from the conditioning electrode when the conditioning and test pulses were delivered simultaneously. This depression decreased when the interval between conditioning and test current pulses was increased, until, at an interval of ∼0.25 ms, it was negligible. At no time during several thousand test-conditioning pairs, for electrodes at different distances apart (28–135 μm) on the same or contiguous terminal branches, did the electrotonic effects of quantal release at one electrode produce quantal release at the other. Analytic and numerical solutions were obtained for the distribution of transmembrane potential at different sites along terminal branches of different lengths for current injection at a point on a terminal branch wrapped in Schwann cell, in the absence of active membrane conductances. Solutions were also obtained for the combined effects of two sites of current injection separated by different time delays. This cable model shows that depolarizing current injections of a few hundred microseconds duration produce hyperpolarizations at ∼30 μm beyond the site of current injection, with these becoming larger and occurring at shorter distances the shorter the terminal branch. Thus the effect of a conditioning depolarizing pulse at one site on a subsequent test pulse at another more than ∼30 μm away is to substantially decrease the absolute depolarization produced by the latter, provided the interval between the pulses is less than a few hundred microseconds. It is concluded that the passive cable properties of motor nerve terminal branches are sufficient to explain the effects on quantal secretion by a test electrode depolarization of current injections from a spatially removed conditioning electrode.

Author(s):  
Norman Robbins ◽  
Joseph Polak

Studies of identified synapses in living animals have revealed ongoing synaptic remodelling in the mature nervous system. We recently demonstrated the existence of filopodia and lamellipodia (structures associated with nerve outgrowth in developing or tissue culture systems) at the mature mouse neuromuscular junction (NMJ) . Evidence of motor nerve terminal retraction, regeneration and perisynaptic outgrowth indicated that even established synaptic contacts were not permanent. It now appears that variation in the stability of synaptic contacts, not the amount of nerve terminal outgrowth,is the basis for synaptic remodelling and extension. Although it has been documented that regenerating motor nerve terminals adhere to former synaptic matrix,and that the matrix contains information for inducing and aligning presynaptic and postsynaptic specializations,it was not clear whether the synaptic matrix was uniformly adhesive, or formed specialized adhesive foci with the preterminal membrane. To determine if there are specific sites of synaptic adhesion we utilized hypertonic fixatives to induce shrinkage at the NMJ.Male mice were anaesthetized, the soleus muscle was exposed, and the overlying skin was retracted to create a pool into which either 2% glutaraldehyde in HEPES Krebs (normal fixative) or 2% glutaraldehyde in HEPES Krebs with a 2X NaCl concentration (hypertonic fixative) was added continually for 30 minutes. The surface layer of each muscle was then removed and submerged in the same fixative for an additional 1.5 hours. Tissue was washed in buffer (pH 7.2) and immersed in cholinesterase stain for 5 minutes to localize endplate regions . Endplate regions were cut into 1 mm blocks, washed in buffer (pH 7.2) and post-fixed for 1 hour in HEPES Krebs buffered 2% osmium tetroxide (pH 7.2). Following fixation, specimens were dehydrated in a graded ethanol series and embedded in Polybed 812 (Polysciences). Mouse soleus muscle was stained for immunocytochemical localization of actin with a rabbit polyclonal antiserum against chicken gizzard actin (from Dr. James Lessard, University of Cincinnati) as previously described .


1991 ◽  
Vol 115 (3) ◽  
pp. 755-764 ◽  
Author(s):  
L Anglister

Acetylcholinesterase (AChE) in skeletal muscle is concentrated at neuromuscular junctions, where it is found in the synaptic cleft between muscle and nerve, associated with the synaptic portion of the myofiber basal lamina. This raises the question of whether the synaptic enzyme is produced by muscle, nerve, or both. Studies on denervated and regenerating muscles have shown that myofibers can produce synaptic AChE, and that the motor nerve may play an indirect role, inducing myofibers to produce synaptic AChE. The aim of this study was to determine whether some of the AChE which is known to be made and transported by the motor nerve contributes directly to AChE in the synaptic cleft. Frog muscles were surgically damaged in a way that caused degeneration and permanent removal of all myofibers from their basal lamina sheaths. Concomitantly, AChE activity was irreversibly blocked. Motor axons remained intact, and their terminals persisted at almost all the synaptic sites on the basal lamina in the absence of myofibers. 1 mo after the operation, the innervated sheaths were stained for AChE activity. Despite the absence of myofibers, new AChE appeared in an arborized pattern, characteristic of neuromuscular junctions, and its reaction product was concentrated adjacent to the nerve terminals, obscuring synaptic basal lamina. AChE activity did not appear in the absence of nerve terminals. We concluded therefore, that the newly formed AChE at the synaptic sites had been produced by the persisting axon terminals, indicating that the motor nerve is capable of producing some of the synaptic AChE at neuromuscular junctions. The newly formed AChE remained adherent to basal lamina sheaths after degeneration of the terminals, and was solubilized by collagenase, indicating that the AChE provided by nerve had become incorporated into the basal lamina as at normal neuromuscular junctions.


Neuron ◽  
1989 ◽  
Vol 3 (6) ◽  
pp. 677-688 ◽  
Author(s):  
Mark Rich ◽  
Jeff W. Lichtman

1990 ◽  
Vol 14 ◽  
pp. 227
Author(s):  
J TOMAS ◽  
R FENOLL ◽  
J BATLLE ◽  
M SANTAFE ◽  
V PIERA ◽  
...  

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