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J Biol Chem, Vol. 275, Issue 11, 7764-7770, March 17, 2000
From the Rho, Rac, and Cdc42 monomeric GTPases are well
known regulators of the actin cytoskeleton and phosphoinositide
metabolism and have been implicated in hormone secretion in endocrine
cells. Here, we examine their possible implication in
Ca2+-dependent exocytosis of
neurotransmitters. Using subcellular fractionation procedures, we found
that RhoA, RhoB, Rac1, and Cdc42 are present in rat brain synaptosomes;
however, only Rac1 was associated with highly purified synaptic
vesicles. To determine the synaptic function of these
GTPases, toxins that impair Rho-related proteins were
microinjected into Aplysia neurons. We used lethal toxin
from Clostridium sordellii, which inactivates Rac; toxin B
from Clostridium difficile, which inactivates Rho, Rac, and Cdc42; and C3 exoenzyme from Clostridium botulinum and
cytotoxic necrotizing factor 1 from Escherichia coli, which
mainly affect Rho. Analysis of the toxin effects on evoked
acetylcholine release revealed that a member of the Rho family, most
likely Rac1, was implicated in the control of neurotransmitter release.
Strikingly, blockage of acetylcholine release by lethal toxin and toxin
B could be completely removed in <1 s by high frequency stimulation of
nerve terminals. Further characterization of the inhibitory action
produced by lethal toxin suggests that Rac1 protein regulates a late
step in Ca2+-dependent neuroexocytosis.
A Rho-related GTPase Is Involved in
Ca2+-dependent Neurotransmitter Exocytosis*
§,,
,,§§
Laboratoire de Neurobiologie
Cellulaire, CNRS, UPR 9009 and ¶ INSERM, U-338 Biologie de la
Communication Cellulaire, F-67084 Strasbourg Cédex,
** Toxines Microbiennes, Institut Pasteur, F-75724 Paris Cédex 15, and INSERM, U-452 Biologie Cellulaire et
Moléculaire des Microorganismes et de leurs Toxines,
F-06107 Nice Cédex 2, France
*
This work was supported by grants from the Association
Française contre les Myopathies and the Programme de Recherche
Fondamentale en Microbiologie et Maladies Infectieuses et Parasitaires
and by Direction des Systèmes des Forces et de la Prospective
Contract 99-34-038 (to B. P.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Present address: Dipt. di Farmacologia e Fisiologia Umana,
Facoltà di Medicina e Chirurgia, Università di Bari,
I-70124 Bari, Italy.
§§
To whom correspondence should be addressed: Lab. de Neurobiologie
Cellulaire, CNRS, 5 rue Blaise Pascal, F-67084 Strasbourg Cédex,
France. Tel.: 33-3-88-45-66-77; Fax: 33-3-88-60-16-64; E-mail:
poulain@neurochem.u-strasbg.fr.
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