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. 2023 Sep 28;12:e86084. doi: 10.7554/eLife.86084

Figure 7. Spinophilin is crucial for aversive olfactory mid-term memories but not for learning.

The Spinophilin/Syd-1 antagonism controls aversive mid-term memories. (a–d) Quantification of BRP and Unc13A intensity in control and Spn knockdown (KD) flies, 1 and 3 h after conditioning. (a,c) BRP and Unc13A levels increase in controls upon paired conditioning while (b,d) BRP and Unc13A do not increase in Spn KD upon paired conditioning. (e) Scheme showing different phases of memory in Drosophila. (f–g) STM is normal in both Spn and Syd-1 KDs. (h) 1 hour and (j) 3 hour MTM are both impaired in Spn KD. (i) 1 hour MTM is normal while (k) 3 hour MTM is impaired in Syd-1 KD. (l) Spn heterozygosity (Spnex3.1/+) shows impaired 3 hr MTM, which is rescued upon concomitant KD of Syd-1. (m) 3 hour MTM impairment in Spn KD is rescued by concomitant reduction in Syd-1 through Syd-1 heterozygosity (Syd-1ex3.4/+). Also see Figure 7—figure supplements 1–3. Source data as exact normalized and raw values, detailed statistics including sample sizes and p values are provided in Figure 7—source data 1. *p≤0.05; **p ≤ 0.01; ***p ≤ 0.001; n.s., not significant, p > 0.05. All panels show mean  ± s.e.m.

Figure 7—source data 1. Table containing exact values for the data depicted in Figure 7, along with details of statistical analyses.

Figure 7.

Figure 7—figure supplement 1. Spinophilin and Syd-1 knockdown results in impaired mid-term memory (MTM), due to impaired anesthesia-sensitive memory (ASM).

Figure 7—figure supplement 1.

(a) Representative Drosophila brain images for data quantified in Figure 7a–d. (c–e) ARM, ASM and 18 °C control performance indices of 1 h MTM in Spn-KD corresponding to Figure 7h. (f–h) ARM, ASM and 18 °C control performance indices of 1 h MTM in Spn-KD corresponding to Figure 7j. (i–k) ARM, ASM and 18 °C control performance indices of 1 hr MTM in Syd-1-KD corresponding to Figure 7k. Source data as exact normalized and raw values, detailed statistics including sample sizes and p values are provided in Figure 7—figure supplement 1—source data 1. *p≤0.05; **p ≤ 0.01; ***p ≤ 0.001; n.s., not significant, p > 0.05. All panels show mean  ± s.e.m.
Figure 7—figure supplement 1—source data 1. Table containining exact values for the data depicted in Figure 7—figure supplement 1, along with details of statistical analyses.
Figure 7—figure supplement 2. The Spinophilin/Syd-1 antagonism is not relevant to 1h mid-term memory (MTM) formation.

Figure 7—figure supplement 2.

(a) Spn heterozygosity (Spnex3.1/+) shows impaired 1 hr MTM, which is not rescued upon concomitant KD of Syd-1. (b) 1 hr MTM impairment in Spn KD is not rescued by concomitant reduction in Syd-1 through Syd-1 heterozygosity (Syd-1ex3.4/+) (c–d) Control experiments at 18 °C show that Syd-1 KD does not rescue Spn heterozygosity and Spn-KD does not show a phenotype at 3hMTM. Source data as exact normalized and raw values, detailed statistics including sample sizes and p values are provided Figure 7—figure supplement 2—source data 1. *p≤0.05; **p ≤ 0.01; ***p ≤ 0.001; n.s., not significant, p > 0.05. All panels show mean  ± s.e.m.
Figure 7—figure supplement 2—source data 1. Table containining exact values for the data depicted in Figure 7—figure supplement 2, along with details of statistical analyses.
Figure 7—figure supplement 3. Model depicting plasticity processes underlying presynaptic homeostatic plasticity in peripheral neuromuscular synapses and memory stabilization in mushroom body KCs.

Figure 7—figure supplement 3.

Spn and Syd-1 antagonism mediates cytoskeletal remodeling which facilitates plasticity responses. Rok replenishes the readily releasable pool of synaptic vesicles (SV) and Spn mediates actin depolymerization which gives SVs access to release sites.