Vesicle exocytosis stimulated by alpha-latrotoxin is mediated by latrophilin and requires both external and stored Ca2+.
alpha-Latrotoxin (LTX) stimulates massive neurotransmitter release by two mechanisms: Ca2+-dependent and -independent. Our studies on norepinephrine secretion from nerve terminals now reveal the different molecular basis of these two actions. The Ca2+-dependent LTX-evoked vesicle exocytosis (abolished by botulinum neurotoxins) is 10-fold more sensitive to external Ca2+ than secretion triggered by depolarization or A23187; it does not, however, depend on the cation entry into terminals but requires intracellular Ca2+ and is blocked by drugs depleting Ca2+ stores and by inhibitors of phospholipase C (PLC). These data, together with binding studies, prove that latrophilin, which is linked to G proteins and inositol polyphosphate production, is the major functional LTX receptor. The Ca2+-independent LTX-stimulated release is not inhibited by botulinum neurotoxins or drugs interfering with Ca2+ metabolism and occurs via pores in the presynaptic membrane, large enough to allow efflux of neurotransmitters and other small molecules from the cytoplasm. Our results unite previously contradictory data about the toxin's effects and suggest that LTX-stimulated exocytosis depends upon the co-operative action of external and intracellular Ca2+ involving G proteins and PLC, whereas the Ca2+-independent release is largely non-vesicular.
Biochemistry Department, Imperial College, Exhibition Road, London SW7 2AY, UK.
B A
BA
Davletov
F A
FA
Meunier
A C
AC
Ashton
H
H
Matsushita
W D
WD
Hirst
V G
VG
Lelianova
G P
GP
Wilkin
J O
JO
Dolly
Y A
YA
Ushkaryov
D000818:Animals; D001905:Botulinum Toxins / Q000494:pharmacology; D019556:COS Cells; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000502:physiology*; D004963:Estrenes / Q000494:pharmacology; D005089:Exocytosis / Q000187:drug effects*; D006023:Glycoproteins; D007476:Ionophores / Q000494:pharmacology; D008297:Male; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D009469:Neuromuscular Junction / Q000378:metabolism; D009479:Neuropeptides; D009638:Norepinephrine / Q000378:metabolism*; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D011760:Pyrrolidinones / Q000494:pharmacology; D011893:Rana esculenta; D051381:Rats; D018000:Receptors, Peptide / Q000378:metabolism*; D012110:Reserpine / Q000494:pharmacology; D013111:Spider Venoms / Q000378:metabolism / Q000494:pharmacology*; D013574:Synaptosomes; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors / Q000502:physiology
D004963:Estrenes; D006023:Glycoproteins; D007476:Ionophores; D009419:Nerve Tissue Proteins; D009479:Neuropeptides; D010726:Phosphodiesterase Inhibitors; D011760:Pyrrolidinones; D018000:Receptors, Peptide; D013111:Spider Venoms; C034800:alpha-latrotoxin receptor; C112162:neurexophilin; C060229:1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione; C096325:botulinum toxin type D; D000001:Calcimycin; C020763:alpha-latrotoxin; D012110:Reserpine; D010738:Type C Phospholipases; D001905:Botulinum Toxins; C096324:botulinum toxin type C; D002118:Calcium; D009638:Norepinephrine