Daily ultrastructural remodeling of clock neurons

In Drosophila, about 250 clock neurons in the brain form a network that orchestrates circadian rhythmicity. Among them, eight small Lateral ventral Neurons (s-LNvs) play a critical role, synchronizing the circadian ensemble via the neuropeptide Pigment-Dispersing Factor (PDF). Moreover, their neurites show daily variations in morphology, PDF levels, synaptic markers and connectivity. This process, called circadian structural plasticity, is ill-defined at the subcellular level. Here, we present 3D volumes of the s-LNv terminals generated by Serial Block-face Scanning Electron Microscopy (SBEM) at three key time points, two hours before lights-ON, two hours after lights-ON, and two hours after lights-OFF. We report a reduction in the number of neuronal varicosities at night, which reflects (and probably regulates) the cycling of the components we found therein. Indeed, in the morning we observed more presynaptic sites and increased accumulation and release of dense core vesicles. These rhythms were paralleled by periodic changes in mitochondrial structure that suggest daily modulation of their activity. We propose that circadian plasticity of the functionally relevant structures within presynaptic varicosities cyclically modulates the influence of the s-LNvs on the clock network.