Implications of the Sap47 null mutation for synapsin phosphorylation, longevity, climbing, and behavioural plasticity in adult Drosophila

Abstract

The Sap47 gene of Drosophila melanogaster encodes a highly abundant 47 kDa synaptic vesicle-associated protein. Sap47 null mutants show defects in synaptic plasticity and larval olfactory associative learning but the molecular function of Sap47 at the synapse is unknown. We demonstrate that Sap47 modulates the phosphorylation of another highly abundant conserved presynaptic protein, synapsin. Site-specific phosphorylation of Drosophila synapsin has repeatedly been shown to be important for behavioural plasticity but it was not known where these phospho-synapsin isoforms are localized in the brain. Here, we report the distribution of serine-6-phosphorylated synapsin in the adult brain and show that it is highly enriched in rings of synapses in the ellipsoid body and in large synapses near the lateral triangle. The effects of knockout of Sap47 or synapsin on olfactory associative learning/memory support the hypothesis that both proteins operate in the same molecular pathway. We therefore asked if this might also be true for other aspects of their function. We show that knockout of Sap47 but not synapsin reduces lifespan, whereas knockout of Sap47 and synapsin, either individually or together, affects climbing proficiency, as well as plasticity in circadian rhythms and sleep. Furthermore, electrophysiological assessment of synaptic properties at the larval neuromuscular junction (NMJ) reveals increased spontaneous synaptic vesicle fusion and reduced paired pulse facilitation in Sap47 and synapsin single and double mutants. Our results imply that Sap47 and synapsin cooperate non-uniformly in the control of synaptic properties in different behaviourally relevant neuronal networks of the fruitfly.

Aversive teaching signals from individual dopamine neurons in larval Drosophila show qualitative differences in their temporal “fingerprint”

“…Regarding the association of odor and electric shock, both forward and backward conditioning are impaired upon of a lack of synapsin (Niewalda et al, 2015). Synapsin is an evolutionarily conserved presynaptic protein with a high number of phosphorylation sites, and consensus motifs for multiple kinases (reviewed in Diegelmann et al, 2013; see also Niewalda et al, 2015, Kleber et al, 2016, Blanco‐Redondo et al, 2019). Synapsin regulates the balance between reserve and readily‐releasable synaptic vesicle pools, and hence synaptic efficacy, across species (Benfenati, 2011; Diegelmann et al, 2013; Hilfiker et al, 1999).…”

“…In an explant brain preparation and with respect to the appetitive domain, Regarding the association of odor and electric shock, both forward and backward conditioning are impaired upon of a lack of synapsin (Niewalda et al, 2015). Synapsin is an evolutionarily conserved presynaptic protein with a high number of phosphorylation sites, and consensus motifs for multiple kinases (reviewed in Diegelmann et al, 2013; see also Niewalda et al, 2015, Kleber et al, 2016, Blanco-Redondo et al, 2019.…”

Central synaptopathy is the most conserved feature of motor circuit pathology across spinal muscular atrophy mouse models

“…The membranes were then incubated with the corresponding antibodies overnight. Thereafter, the membranes were incubated with IRDye 680RD or 800CW secondary antibodies (Li-cor) followed by visualization using a near-infrared imager (Odyssey; Li-cor) ( Blanco-Redondo et al, 2019 ). Band intensities were analyzed by Fiji ImageJ.…”

Chronic Pharmacological Increase of Neuronal Activity Improves Sensory-Motor Dysfunction in Spinal Muscular Atrophy Mice

“…The membranes were then incubated with the corresponding antibodies overnight. Thereafter, the membranes were incubated with IRDye 680RD or 800CW secondary antibodies (LI-COR) followed by visualization using a near-infrared imager (Odyssey; LI-COR; Blanco-Redondo et al, 2019).…”