Membrane recycling due to low and high rates of nerve stimulation at release sites in the amphibian (Bufo marinus) neuromuscular junction.
The activity-dependent labelling of motor nerve terminals with the dye FM1-43 has been used to estimate the relative levels of membrane recycling (due to synaptic vesicle exocytosis and recovery) at release sites in response to 1,200 nerve stimulations delivered at either low (0.5 Hz) or high (30 Hz) frequency. Dye in terminals appears as fluorescent spots distributed along the terminal branches; each spot is thought to be a cluster of labelled vesicles associated with a release site. Relative fluorescence in spots was quantified from images obtained with a confocal microscope. Spot intensities varied widely within branches following labelling at both frequencies, but the distribution was highly skewed towards lower intensities at low frequency stimulation; at high frequency, more spots had stronger fluorescence. Both weak and strongly stained spots were uniformly distributed along the length of terminal branches after low frequency stimulation; however, there was a gradual decline in all spot intensities towards the distal end of branches loaded with dye at high frequency stimulation. Antibody staining for synaptic vesicles was, on average, uniformly distributed along the branches. The increase in number of more strongly FM1-43-labelled spots in terminal branches stimulated at high compared with low frequency suggests that more release sites are active at high rates of nerve stimulation. This "recruitment" of release sites at high frequency stimulation occurs mostly in the proximal half of terminal branches and is not related to the abundance of synaptic vesicles in the terminal.
Everett
A W
AW
Department of Physiology, The University of Western Australia, Nedlands, Australia. aeverett@cyllene.uwa.edu.au
Packard
S J
SJ
Cosby
M
M
Milne
R K
RK
D000818:Animals; D002024:Bufo marinus; D005089:Exocytosis / Q000502:physiology; D005456:Fluorescent Dyes; D008565:Membrane Proteins / Q000378:metabolism; D018613:Microscopy, Confocal; D009046:Motor Neurons / Q000737:chemistry / Q000378:metabolism*; D009469:Neuromuscular Junction / Q000737:chemistry / Q000378:metabolism*; D011726:Pyridinium Compounds; D000644:Quaternary Ammonium Compounds; D009435:Synaptic Transmission / Q000502:physiology*; D013572:Synaptic Vesicles / Q000737:chemistry / Q000378:metabolism
C073804:FM1 43; D005456:Fluorescent Dyes; D008565:Membrane Proteins; D011726:Pyridinium Compounds; D000644:Quaternary Ammonium Compounds