Clathrin-mediated endocytosis in snake motor terminals is directly facilitated by intracellular Ca2+.
At the snake neuromuscular junction, low temperature (LT, 5-7 degrees C) blocks clathrin-mediated endocytosis (CME) while exocytosis is largely unaffected. Thus compensatory endocytosis that normally follows transmitter release is inhibited, or 'delayed' until the preparation is warmed to room temperature (RT). This delay was exploited to observe how changes in bulk [Ca(2+)](i) directly affect CME. Motor terminals were loaded with fura-2 to monitor [Ca(2+)](i). With brief stimulation at LT, [Ca(2+)](i) transiently increased but returned to baseline ( approximately 63 nm) in < 8 min. After 15 min at LT, [Ca(2+)](i) was altered by incubating preparations in the Ca(2+) ionophore ionomyocin. Preparations were then warmed to RT to initiate delayed endocytosis, which was quantified as uptake of the fluorescent optical probe sulforhodamine 101. Endocytosis was more rapid when [Ca(2+)](i) increased; the rate at 300 nm Ca(2+) was approximately double that under basal conditions. Thus the rate of CME - isolated from stimulation, transmitter release, and other forms of endocytosis - is directly influenced by intraterminal Ca(2+).
Teng
Haibing
H
Department of Cell Biology and Physiology, Washington University School of Medicine, 660 South Euclid Ave., Box 8228, St. Louis, MO 63110, USA.
Wilkinson
Robert S
RS
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002966:Clathrin / Q000378:metabolism*; D003080:Cold Temperature; D017812:Colubridae; D004558:Electric Stimulation; D004705:Endocytosis / Q000187:drug effects / Q000502:physiology*; D005089:Exocytosis / Q000502:physiology; D005456:Fluorescent Dyes; D016257:Fura-2; D015759:Ionomycin / Q000494:pharmacology; D007476:Ionophores / Q000494:pharmacology; D009469:Neuromuscular Junction / Q000502:physiology*
D002966:Clathrin; D005456:Fluorescent Dyes; D007476:Ionophores; D015759:Ionomycin; D002118:Calcium; D016257:Fura-2