Serotonin released within the dorsal raphe nucleus (DR) induces feedback inhibition of serotonin neuron activity and consequently regulates mood-controlling serotonin release throughout the forebrain. Serotonin packaged in vesicles is released in response to action potentials by the serotonin neuron soma and terminals, but the potential for release by dendrites is unknown. Here three-photon (3P) microscopy imaging of endogenous serotonin in living rat brain slice, immunofluorescence and immuno-gold electron microscopy detection of VMAT2 (vesicular monoamine transporter 2) establish the presence of vesicular serotonin within DR dendrites. Furthermore, activation of glutamate receptors is shown to induce vesicular serotonin release from dendrites. However, unlike release from the soma and terminals, dendritic serotonin release is independent of action potentials, relies on L-type Ca2+ channels, is induced preferentially by NMDA, and displays distinct sensitivity to the selective serotonin reuptake inhibitor (SSRI) antidepressant fluoxetine. The unique control of dendritic serotonin release has important implications for DR physiology and the antidepressant action of SSRIs, dihydropyridines and NMDA receptor antagonists.

Peptide release from single dense-core vesicles (DCVs) was detected at a native nerve terminal for the first time. Synaptic release is slow, dynamin dependent, and mediated exclusively by kiss-and-run exocytosis. Presynaptic peptide release is independent of DCV arrival time, bouton location, and transport direction into the synapse.

Synaptic neuropeptide and neurotrophin stores are maintained by constitutive bidirectional capture of dense-core vesicles (DCVs) as they circulate in and out of the nerve terminal. Activity increases DCV capture to rapidly replenish synaptic neuropeptide stores following release. However, it is not known whether this is due to enhanced bidirectional capture. Here experiments at the Drosophila neuromuscular junction, where DCVs contain neuropeptides and a bone morphogenic protein, show that activity-dependent replenishment of synaptic neuropeptides following release is evident after inhibiting the retrograde transport with the dynactin disruptor mycalolide B or photobleaching DCVs entering a synaptic bouton by retrograde transport. In contrast, photobleaching anterograde transport vesicles entering a bouton inhibits neuropeptide replenishment after activity. Furthermore, tracking of individual DCVs moving through boutons shows that activity selectively increases capture of DCVs undergoing anterograde transport. Finally, upregulating fragile X mental retardation 1 protein (Fmr1, also called FMRP) acts independently of futsch/MAP-1B to abolish activity-dependent, but not constitutive, capture. Fmr1 also reduces presynaptic neuropeptide stores without affecting activity-independent delivery and evoked release. Therefore, presynaptic motoneuron neuropeptide storage is increased by a vesicle capture mechanism that is distinguished from constitutive bidirectional capture by activity dependence, anterograde selectivity, and Fmr1 sensitivity. These results show that activity recruits a separate mechanism than used at rest to stimulate additional synaptic capture of DCVs for future release of neuropeptides and neurotrophins.

The Kv4.3 transient outward current (Ito) channel, which produces early repolarization in human cardiomyocytes, is downregulated with cardiac pathology. This is evident in cultured neonatal rat cardiomyocytes in which Angiotensin II (Ang II) acts via p38 mitogen-activated protein kinase (p38K) to increase apoptosis and induce Kv4.3 mRNA destabilization to downregulate the channel protein. However, it is not understood how p38K activation, which is activated transiently for minutes, induces downstream effects hours later. Here we show that there is a second phase of p38K activation. Inhibiting this delayed p38K activation eliminated Kv4.3 mRNA destabilization. Furthermore, inhibiting endosome generation left the transient activation of p38K intact, but blocked delayed p38K activation and the Kv4.3 effect. CamKII was also found to be required for delayed p38K activation and Kv4.3 mRNA destabilization. Finally, CamKII methionine oxidation and activation are biphasic, with the delayed phase requiring endosomes. Hence, in addition to participating in channel traffic, cardiomyocyte endosomes control channel mRNA expression by mediating delayed oxidative CamKII-p38K signaling.

Although dynactin was believed to be a bidirectional facilitator of axonal transport, here mycalolide B is identified as a dynactin dissociator and shown to selectively abolish retrograde axonal transport of dense-core vesicles in hippocampal and Drosophila neurons. Thus dynactin has a strict obligatory unidirectional role in axonal transport.