Deletion of synaptotagmin-1 (Syt1) in forebrain neurons blocks fast synchronous release induced by isolated action potentials but retains an asynchronous,

slower, and facilitating form of release induced by bursts of action potentials (Geppert et al., 1994, Yoshihara and Littleton, 2002, Maximov and Südhof, 2005 and Xu et al., 2012). Although in most synapses asynchronous release becomes manifest only when Syt1 is deleted, in some synapses asynchronous release normally predominates. This is observed in GABAergic synapses formed by CCK-containing Nutlin-3 manufacturer dentate gyrus interneurons (Hefft and Jonas, 2005 and Daw et al., 2009) and inferior olive interneurons (Best and Regehr, 2009). The mammalian genome encodes 16 synaptotagmins, eight of which bind Ca2+ (reviewed in Gustavsson and Han, 2009). Of these eight synaptotagmins, Syt1, Syt2, and Syt9 are localized to synaptic and neuroendocrine vesicles and function as Ca2+ sensors for fast synaptic and neuroendocrine

exocytosis (Perin et al., 1990, Brose et al., 1992, Littleton et al., 1993, Geppert et al., 1994, Sørensen et al., 2003, Pang et al., 2006, Sun et al., 2007 and Xu et al., 2007). Syt10, in contrast, is localized to IGF-1-containing vesicles in olfactory bulb neurons and acts as Ca2+ sensor for IGF-1 secretion in these neurons (Cao et al., 2011 and Cao et al., 2013). Among the remaining four Ca2+-binding synaptotagmins, Syt7 is particularly interesting because it is highly expressed in neurons and enriched Thiamine-diphosphate kinase in synapses (Sugita et al., 2001 and Virmani check details et al., 2003). Surprisingly, Syt7 is dispensable for neurotransmitter release in cultured neurons (Maximov et al., 2008), although it contributes to asynchronous release in zebrafish neuromuscular junctions (Wen et al., 2010). At synapses, Syt7 was not detected in synaptic vesicles but in the synaptic plasma membrane, whereas Syt1 was found in synaptic

vesicles (Sugita et al., 2001, Han et al., 2004, Takamori et al., 2006 and Maximov et al., 2007). In neuroendocrine cells, however, Syt7 was colocalized with Syt1 on secretory granules and was shown to mediate Ca2+ triggering of exocytosis similar to Syt1 (Sugita et al., 2001, Shin et al., 2002, Fukuda et al., 2004, Tsuboi and Fukuda, 2007, Schonn et al., 2008, Gustavsson et al., 2008, Gustavsson et al., 2009, Li et al., 2009 and Segovia et al., 2010), albeit with a slower time course (Schonn et al., 2008). Thus, Syt7 is an evolutionarily conserved synaptotagmin highly expressed in brain that functions in neuroendocrine exocytosis but whose neural function is unclear. Despite its importance, the identity of the Ca2+ sensor mediating asynchronous release that becomes manifest in Syt1-deficient synapses has remained enigmatic. One study implicated Doc2A as a Ca2+ sensor for asynchronous release (Yao et al., 2011), but other studies failed to detect any role for Doc2 proteins in asynchronous release (Groffen et al., 2010 and Pang et al., 2011a).