L-type calcium channels are involved in fast endocytosis at the mouse neuromuscular junction.
We used fluorescence microscopy of FM dyes-labeled synaptic vesicles and electrophysiological recordings to examine the functional characteristics of vesicle recycling and study how different types of voltage-dependent Ca(2+) channels (VDCCs) regulate the coupling of exocytosis and endocytosis at mouse neuromuscular junction. Our results demonstrate the presence of at least two different pools of recycling vesicles: a high-probability release pool (i.e. a fast destaining vesicle pool), which is preferentially loaded during the first 5 s (250 action potentials) at 50 Hz; and a low-probability release pool (i.e. a slow destaining vesicle pool), which is loaded during prolonged stimulation and keeps on refilling after end of stimulation. Our results suggest that a fast recycling pool mediates neurotransmitter release when vesicle use is minimal (i.e. during brief high-frequency stimulation), while vesicle mobilization from a reserve pool is the prevailing mechanism when the level of synaptic activity increases. We observed that specific N- and L-type VDCC blockers had no effect on evoked transmitter release upon low-frequency stimulation (5 Hz). However, at high-frequency stimulation (50 Hz), L-type Ca(2+) channel blocker increased FM2-10 destaining and at the same time diminished quantal release. Furthermore, when L-type channels were blocked, FM2-10 loading during stimulation was diminished, while the amount of endocytosis after stimulation was increased. Our experiments suggest that L-type VDCCs promote endocytosis of synaptic vesicles, directing the newly formed vesicles to a high-probability release pool where they compete against unused vesicles.
Perissinotti
Paula P
PP
Departmento de Fisiología Biología Molecular y Celular, Universidad de Buenos Aires, Instituto de Fisiología Biología Molecular y Neurociencias, UBA CONICET, Argentina.
Giugovaz Tropper
Bárbara
B
Uchitel
Osvaldo D
OD
D000818:Animals; D002121:Calcium Channel Blockers / Q000494:pharmacology; D020746:Calcium Channels, L-Type / Q000502:physiology*; D004558:Electric Stimulation / Q000379:methods; D004705:Endocytosis / Q000187:drug effects / Q000502:physiology*; D008297:Male; D051379:Mice; D009469:Neuromuscular Junction / Q000187:drug effects / Q000502:physiology*; D017729:Presynaptic Terminals / Q000187:drug effects / Q000502:physiology; D013572:Synaptic Vesicles / Q000187:drug effects / Q000502:physiology; D013997:Time Factors
D002121:Calcium Channel Blockers; D020746:Calcium Channels, L-Type