, 2005, Madison et al., 2005, Stevens et al., 2005 and Guan et al., 2008), but the mechanisms of Munc13 function in priming, and of the inactivation of Munc13 function by homodimerization, remain unclear. One possibility is that homodimeric Munc13 is inherently unstable and becomes degraded in RIM-deficient neurons, thereby accounting for the priming phenotype and the reduced Munc13 levels in RIM-deficient neurons (Figure 1; Schoch et al., 2002). However, overexpression of wild-type Munc13 did not rescue the priming phenotype in RIM-deficient neurons, suggesting that simply increasing Munc13 levels is not sufficient to rescue priming in RIM-deficient synapses. Another possibility is that homodimeric Munc13 is not

correctly targeted to synapses and becomes degraded if it is not in the correct location (Andrews-Zwilling et al., 2006 and Kaeser et al., 2009). Although

possible, this hypothesis check details appears rather unlikely given the rescue of the RIM- and Munc13-deficiency phenotypes by N-terminally truncated Munc13 (Figure 7 and Figure 8), which suggests that Munc13 is transported to synapses without RIM proteins and without binding to RIM proteins. Independent of which explanation will turn out to be correct, the mechanism of Munc13 activation we identify here is opposite to what is classically observed for signal transduction events; dimerization Dactolisib mouse is usually activating, whereas in our case it is inhibitory, suggesting a more diverse range of biological activation mechanisms than previously envisioned. The current study identifies a molecular mechanism involved in vesicle priming by the active zone but raises new questions. At a basic level, how is an active zone generated—what protein nucleates its assembly? The fact that the RIM Zn2+ finger alone is active suggests that it acts downstream of Munc13 targeting to active zones and cannot physically tether Munc13 to them; similarly, Munc13

is not essential for targeting other proteins to active zones and thus also not secondly involved in their recruitment to active zones. Clearly, despite its central function, RIM alone does not organize the active zone, an activity that may be carried out by an overlapping set of several proteins instead of a single master regulator. Another important question is how RIM proteins contribute to long-term synaptic plasticity—is this mediated by a coordination of their various functions or by one particular aspect? With the present results, we now know of two switches at the active zone that involve RIM and regulate synaptic neurotransmitter release: the GTP-dependent interaction of Rab3 with RIMs, and the Zn2+ finger mediated RIM-dependent monomerization of Munc13. Given the central roles of RIM and Rab3 in all known forms of long-term presynaptic plasticity (e.g., Castillo et al., 1997, Castillo et al., 2002, Chevaleyre et al., 2007, Fourcaudot et al., 2008 and Kaeser et al.