, 2002, Majewska et al., 2000a and Sabatini et al., 2002). Calcium compartmentalization by spines could allow long-term synaptic plasticity at individual synaptic sites (Holmes,

1990, Koch and Zador, 1993 and Malenka et al., 1988). Indeed, very high spine calcium accumulations are triggered by stimulation protocols that generate LTP (Koester and Sakmann, 1998 and Yuste et al., 1999). Moreover, the increase in synaptic strength after LTP is accompanied by a corresponding increase in the volume of the spine head (Matsuzaki et al., 2004), and selleck chemicals llc this volume is proportional to the size of the PSD and the number of glutamate receptors in it (Arellano et al., 2007a, Harris et al., 1992 and Schikorski and Stevens, 1999). All of these separate pieces of evidence are consistent with a model by which the stimulation of an individual spine, when paired with backpropagating action potentials, triggers a calcium influx specific to the activated spine and elicits LTP by inserting glutamate receptors into that synapse, without affecting the neighboring synapses. Besides this

biochemical compartmentalization, there is an additional mechanism by which spines could enable input-specific alterations in synaptic strength. If the spine neck has a significant resistance, as discussed above, changes in its length or width, or in its electrical properties that may not be morphologically detectable, could alter synaptic strength. This idea, first

proposed by Rall (Rall, 1974a and Rall, Metformin 1995), has become more tenable through the realization that spines are not rigid structures but can dynamically alter their shape and length, in a matter of seconds (Dunaevsky et al., 1999 and Fischer et al., 1998). In fact, significant alterations in the dimensions of the spine neck occur spontaneously (Dunaevsky et al., 1999, Majewska et al., 2000b and Parnass et al., 2000) and changes in spine neck diffusion occurs in response to synaptic activity (Bloodgood and Sabatini, 2005). Moreover, electron microscopic reconstructions indicate that the spine neck becomes shorter whatever and wider after LTP (Fifková and Anderson, 1981 and Fifková and Van Harreveld, 1977), potentially explaining the increase of synaptic strength. These neck-based changes in synaptic strength could be fast and would not require altering the number of synaptic receptors, but merely alter the spine’s electrical coupling to the dendrite. Finally, there is a third mechanism by which spines provide enhanced synaptic plasticity. As mentioned above, by specifically enabling connections with a larger variety of axons, spines could allow rewiring that would be much more extensive than if synapses were on dendritic shafts and were to contact only a limited assortment of axons (Chklovskii et al., 2002).

By contrast, when either FLP-18 or FLP-21 was inactivated in a selleckchem Bristol strain expressing low-affinity NPR-1(215F) receptors, i.e., npr-1(g320) mutants, locomotion quiescence was significantly decreased ( Figures 1F and 1G). These results suggest that FLP-18 and FLP-21 function as endogenous NPR-1 ligands to regulate lethargus behavior in strains expressing NPR-1(215F) receptors. NPR-1’s effects on foraging are mediated by its expression in a sensory circuit in the head

that is defined by gap junctions to the RMG interneuron (Figure 2A) (Macosko et al., 2009). Hereafter, we refer to this circuit as the RMG circuit. In addition to the RMG circuit, NPR-1 is also expressed in GABAergic motor neurons in the ventral nerve cord (Coates and de Bono, 2002).

We did two experiments to determine where NPR-1 functions to regulate motility Hydroxychloroquine molecular weight during lethargus. First, an npr-1 transgene expressed in the RMG circuit (using the flp-21 promoter) ( Figure 2A) completely rescued the lethargus locomotion defect of npr-1 mutants, whereas a transgene expressed in GABAergic motor neurons (using the unc-30 promoter) had no rescuing activity ( Figures 2B–2D). Second, the lethargus locomotion defect of npr-1 mutants was abolished by mutations inactivating ion channels required for sensory transduction, such as TAX-4/CNG and OSM-9/TRPV channels ( Figures 2E–2G, S2A, and S2B). A transgene expressing TAX-4 in the RMG circuit reinstated the L4/A quiescence defect in tax-4; npr-1 double mutants ( Figures 2F and 2G). These results suggest that the npr-1 defect in locomotion quiescence during lethargus was caused by heightened sensory activity in the RMG circuit. Neuropeptides play a pivotal role in sleep and wakefulness in Dichloromethane dehalogenase other

systems. For example, hypocretin/orexin regulates sleep, arousal, feeding, and metabolism in vertebrates (Sutcliffe and de Lecea, 2002). Thus, we tested whether neuropeptides are required for the npr-1 lethargus defect. Consistent with this idea, the npr-1 lethargus quiescence defect was eliminated by mutations inactivating egl-3 PC2 and pkc-1 PKCε ( Figures 3A–3C), which are required for proneuropeptide processing and dense-core-vesicle (DCV) exocytosis, respectively ( Husson et al., 2006; Kass et al., 2001; Sieburth et al., 2007). These results suggest that the npr-1 lethargus defect was mediated by an endogenous neuropeptide. In Drosophila, the neuropeptide PDF regulates circadian rhythms and promotes wakefulness ( Parisky et al., 2008; Renn et al., 1999). Prompted by PDF’s role in Drosophila, we tested the idea that PDF mediates the lethargus quiescence defect in npr-1 mutants. C. elegans PDF peptides (PDF-1 and PDF-2) and their receptor (PDFR-1) were previously identified ( Janssen et al., 2008, 2009). PDF-1 is expressed in several classes of sensory neurons and interneurons, including ASK chemosensory neurons and RMG interneurons in the RMG circuit ( Barrios et al.

A similar finding of

altered APP localization to endosomes was also described in the context of iPSC-derived neurons from familial AD associated with a duplication of the APP locus (Israel et al., 2012). Of note, the APP cellular relocalization phenotype is not simply a secondary effect of increased Aβ production, as pharmacological blockade of APP processing failed to suppress the modified APP localization. Genetic “rescue” studies, in which wild-type PSEN1 overexpressed in the PSEN1 mutant hiN cultures suppressed the disease-associated Selleckchem Regorafenib phenotypes, support a direct role for PSEN1 mutation in the phenotype of PSEN1 mutant cells, rather than a spurious effect due to unrelated common variants that may be present in these cultures ( Qiang

et al., 2011). These initial studies with human neuron models of familial AD supported the notion that processes other than extracellular Aβ fragment accumulation may play a role in AD pathology. To further address this, Kondo et al. (2013) used human iPSC-derived forebrain cortical neurons that harbor an APP mutation, V717L, also associated with a familial clinical dementia syndrome of the Alzheimer’s type, Fulvestrant price but one that appears to lack the typical amyloid plaques, composed largely of extracellular Aβ42. iPSC-derived neurons from patients with the V717L APP mutation showed reduced extracellular Aβ42 and Aβ40, consistent with the CNS pathology in human patients with this mutation. Interestingly, intracellular accumulation of Aβ forms was increased (Kondo et al., 2013), suggesting an alternative mechanism of pathology. The increase intracellular Aβ was correlated with markers of endoplasmic reticulum (ER)

and oxidative stress, as well as apoptosis, in the iPSC-derived neuron cultures carrying the V717L mutation. Docosahexaenoic acid (DHA), a therapeutic candidate for AD, relieved the ER stress responses and suppressed apoptosis in the mutant cells. An additional pathological finding that typifies AD patient brain is the accumulation of modified, hyperphosphorylated, and aggregated TAU protein, leading to the L-NAME HCl accumulation of neurofibrillary tangles. APP mutant human iPSC-derived neuron cultures have been reported to harbor increased TAU phosphorylation (Israel et al., 2012), whereas the majority of transgenic rodent models fail to do so, likely reflecting species differences in the TAU gene. Interestingly, inhibition of γ-secretase—which is required for Aβ fragment generation—failed to suppress such phospho-TAU pathology in iPSC-derived APP mutant neurons, whereas inhibition of β-secretase function appeared effective ( Israel et al., 2012). As inhibition of either secretase complex suppresses Aβ production, this finding further supported the notion that aspects of APP biology other than extracellular Aβ accumulation may play an important role in AD pathology.

We considered the trace starting from the beginning of the horizontal line and ending at the determined trans-isomer molecular weight end time. We measured the amplitude from the horizontal line to the peak of the interpolated trace, measured the width of the interpolated trace at the Vm halfway between the horizontal line and the peak (crossing any transient dips between the outermost limits), and computed the product of this amplitude and width. For each cell, we set three thresholds for amplitude, width, and amplitude × width, and classified those events that satisfied all three thresholds to be CSs. The thresholds were ∼15 mV, ∼25 ms,

and ∼15 × 25 mV × ms, respectively, adjusted manually based on visual inspection of the resulting classification. A few events classified as CSs were rejected upon manual inspection. For determining the location at which a CS occurred, we set the time of occurrence to be that of the peak of the first AP in the CS. The Vm reached by the slow, large depolarization of each CS was determined as follows. Intervals from the minimum Vm between 3 ms before the peak and the peak, to the minimum Vm between the peak and 5 ms after the peak of each AP MK0683 chemical structure or spikelet were removed from

the CS’s Vm trace, linear interpolation was applied across the resulting gaps, the interpolated trace was low-pass-filtered with high cutoff 20 Hz, then the peak was taken from this smoothed trace. Figure S2A shows the distribution of these values for all CSs from all place and silent cells (mean ± SD = −24.2 ± 4.4 mV). The mean plateau level of all CSs from a given cell was consistent across place, silent, active, and nonactive

cells (mean ± SD = −24.1 ± 2.8 mV) (only one silent cell and no additional nonactive cells had CSs). This mean plateau level and the baseline Vm were uncorrelated (ρ = −0.12; p = 0.76; regression line: mean CS plateau level = −0.078 × baseline Vm − 29.0 mV), and the mean plateau level and AP threshold were uncorrelated (ρ = 0.41; p = 0.27; mean CS plateau level = 0.26 × AP threshold − 10.4 mV) across cells. Immediately upon breaking into the neuron and achieving the whole-cell recording configuration, while the animal was anesthetized, we injected mafosfamide a series of depolarizing current steps. For each step, the current started at 0 nA, lasted for 300 ms, then returned to zero. The first depolarizing step was 0.1 or 0.2 nA and was increased in increments of 0.1 or 0.2 nA, respectively, for successive steps. The firing pattern of the first step that evoked ≥5 APs was used to determine the propensity to burst and is shown for each cell in Figure 5. The degree of bursting was defined as the fraction of all APs in the firing pattern that occurred in bursts of ≥2 APs with ISIs ≤10 ms.

To verify that the LTD that occurs Raf inhibitor when cAMP/PKA signaling is inhibited is actually eCB-LTD and not another form of synaptic plasticity, we added the CB1 receptor antagonist AM251 to an extracellular recording solution, which already contained CGS21680 and patched cells with PKI included in the intracellular solution. The addition of AM251 to the extracellular solution blocked LTD (119% ± 16%; p < 0.05 compared to LTD with CGS21680 and PKI; Figure S2D), demonstrating that cAMP/PKA inhibition is allowing eCB-LTD to occur. To further test the hypothesis that increases in cAMP/PKA signaling are sufficient to block LTD, we tested

whether directly activating either adenylyl cyclase or PKA would Alpelisib block HFS-LTD when there were no drugs present in the external saline solution. To activate adenylyl cyclase we used the water-soluble (membrane-impermeable) forskolin analog NKH477. To activate PKA we used a membrane-impermeable PKA activator, Sp-8-OH-cAMPS. When either NKH477 or Sp-8-OH-cAMPS were included in

our intracellular recording solution, LTD was inhibited (78% ± 5% with NKH477; 89% ± 9% with Sp-8-OH-cAMPS; both p < 0.05 compared to control LTD; Figures 5C and 5D). From these experiments, we concluded that increased cAMP/PKA activity inhibits LTD. How does cAMP/PKA activity block LTD? Because D2 and A2A receptor drugs act on both LFS- and HFS-LTD, they likely act on a common target in both pathways: group I mGluRs or Gq. A particularly attractive candidate for such modulation is regulator of G protein signaling 4 (RGS4). RGS4 is a GTPase-activating protein expressed strongly in MSNs in the dorsolateral striatum, where it is associated with mGluR5 and PLCβ (Gold et al., 1997 and Schwendt and McGinty, 2007), its activity is increased by tuclazepam PKA phosphorylation (Huang et al., 2007), and it strongly inhibits signaling through Gq (Saugstad et al.,

1998). To test whether RGS4 is involved in the regulation of LTD by D2 and A2A receptors, we obtained RGS4−/− mice, crossed those mice into our D2-EGFP BAC transgenic line so that we could identify indirect-pathway MSNs, and applied the HFS-LTD induction protocol to indirect-pathway MSNs. In RGS4−/− mice, we observed significant HFS-LTD (66% ± 5%; Figure 6A). If RGS4 is responsible for the connection between D2 and A2A receptor signaling and Gq signaling, then LTD in RGS4−/− mice should occur even in the presence of the D2 antagonist sulpiride or the A2A agonist CGS21680. Indeed, LTD was readily observed in the presence of either drug in RGS4−/− mice (80% ± 6% in sulpiride; 64% ± 7% in CGS21680; Figure 6B). While genetic knockouts offer complete elimination of the gene product, they also may yield developmental and/or homeostatic changes. Therefore, we used a recently identified small-molecule inhibitor of RGS4, CCG-63802 (Blazer et al., 2010), to test whether acute inhibition of RGS4 could uncouple LTD from D2 receptors.

Only a large-scale prospective study, as we previously conducted for MDMA (de Win et al., 2008), will be able to show the causal nature of our findings, and exclude that pre-existing differences (such as low D2 receptor availability) underlie our findings. Addiction has also long been associated with aberrant reward-related Z-VAD-FMK molecular weight responses (for a review see Volkow et al., 2011). It has been demonstrated that alcoholics show reduced ventral striatum activation during the anticipation of monetary gain (Wrase et al., 2007)

and a correlation between this response and impulsivity measures has also been reported (Beck et al., 2009). However, cocaine dependent were not different from healthy controls in the anticipation of reward (Asensio et al., 2010). Therefore addiction alone cannot be held exclusively responsible for the changes in reward-related behavior. Although we cannot exclude that the participants in our study were addicted, they clearly stated that they were recreational dAMPH users and not diagnosed with addiction or substance abuse in the past. Moreover, they used dAMPH “only” 28 times per year, which is about once every two weeks and this can

hardly be called addictive use of dAMPH with loss of control. Therefore, it is unlikely that addiction-related changes in the mesolimbic DA pathway involved in drug-reward selleck inhibitor are the predominant mechanism underlying our results. Another explanation for the reduced sensitivity for reward in the recreational dAMPH users is that this is caused by neurotoxic changes induced by chronic dAMPH use. This interpretation

is based on a large body of preclinical studies, such as that from Ricaurte et al. (2005) who observed a reduction in the number of both DAT and vesicular science monoamine transporter (VMAT) in non-human primates treated with a dAMPH in a regimen similar to the one used in the treatment of patients with ADHD (Ricaurte et al., 2005). In addition, PET studies in amphetamine treated vervet monkeys have shown reductions in striatal [18F]fluoro-l-dopa uptake (Melega et al., 1996 and Melega et al., 1997) and reductions in DAT have been observed in combined dAMPH and MDMA users using [123I]β-CIT SPECT (Reneman et al., 2002). Furthermore, studies on the striatal DAergic system in rats have shown that chronic dAMPH exposure results in neurotoxicity characterized by decreases in DA levels and DAT densities, swollen nerve terminals and degenerated axons (Ricaurte et al., 1984). Given the large body of evidence directly documenting the DAergic neurotoxic potential of dAMPH in rodents and nonhuman primates, and because reward functions are strongly connected to the DA system, our data provide further evidence that recreational use of dAMPH is associated with DAergic dysfunction, as evidenced by a reduced activation during reward anticipation.

This is particularly important

for samples containing low Listeria spp. numbers. Indeed, Listeria spp. grow slower than Enterobacteriaceae and its growth could be inhibited by the beef carcass co-resident microflora, and by Salmonella spp. in case of double contamination and the pre-enrichment in BPW instead of Half-Fraser is less optimal for Listeria spp. growth. However, these results demonstrated that BPW is efficient enough for Listeria detection as low as 4–16 cfu/swab with an overnight storage of the swab samples at fridge temperature. From these results, the positive and negative agreements (PA and NA), the positive and negative deviations (PD and ND) were assessed. For both targets, the PA and the NA result were 12 and 8 respectively while the ND and PD were 0. These values allowed the calculation of a 100% relative sensitivity (SE), 100% relative specificity Akt inhibitor (SP) and 100% relative accuracy (AC) and a Cohen’s kappa index of 1.00 of the results obtained with the complete CoSYPS Path Food workflow compared with the results obtained with the ISO methods (Table 2). These values demonstrated that the complete CoSYPS Path Food workflow is as efficient as the reference methods in detecting Salmonella spp. and L. monocytogenes in beef carcass swab samples. click here To perform the ISO reference methods, as well as the complete CoSYPS Path Food

workflow, classical L2 laboratory microbiological equipments are required. In addition, the complete CoSYPS Path Food workflow required qPCR well-trained personnel operating a properly maintained qPCR apparatus. The ISO 11290-1:1996 and ISO

6579:2002 comprise a pre-enrichment step, a selective enrichment step and isolation on selective plates (Fig. 1). These different steps need four and five days for Salmonella spp. and L. monocytogenes detection, respectively, since each culture step requires an 18 to 24 h of incubation (48 h for Fraser selective enrichment). If no typical colonies are observed on selective plates, the sample is concluded as containing no Salmonella spp. or L. monocytogenes all and the analysis is stopped. If typical colonies are observed on the selective plates, it is a presumptive positive result, and further biochemical confirmations are performed, which takes an additional day ( Fig. 1). The complete CoSYPS Path Food workflow comprises a pre-enrichment step, followed by a DNA extraction and a CoSYPS detection system (qPCR analysis). These steps can be completed within two days (including an overnight enrichment) as DNA extraction and CoSYPS analysis are easily performed within a single day. Indeed, DNA extraction requires maximum 3 h and the CoSYPS analysis needs around 4 h (preparation, running and result interpretation). If the CoSYPS analysis result is negative, the sample is concluded as containing neither Salmonella spp. nor Listeria spp. and the analysis is stopped.

By applying a conservative threshold of <0.4, we identified voxels that either are strongly orientation selective or have no selectivity (responded to all orientations).

Using 1 − circular variance to examine preferred orientation on a voxel-by-voxel basis reveals clustering of voxels according to motion preference as derived from the calculated complex angle (Figure 3B). Polar plots from individual voxels illustrate such highly orientation-selective selleck chemicals responses (Figure 3C). Examining the cumulative distribution of complex angles across all larvae imaged reveals two clear populations (Figure 3D) plus a baseline component that reflects the voxels responding to all orientations with noise randomly and evenly distributing the calculated complex angles. Iteratively fitting two summed von-Mises distributions (constrained with bimodal distributions separated JQ1 purchase by 180° and equal concentration) plus a baseline component to the histogram data derived distinct population peaks centered at 105°/285° and 172°/352° (Figure 3D).

These correspond to motion of vertically oriented bars moving along the horizontal axis (horizontally tuned) and horizontally oriented bars moving along the vertical axis (vertically tuned), respectively (Figure 3G). The largest fraction of orientation-selective voxels is tuned to vertical motion. Within all individual larvae examined, the relative proportions of voxels selective for vertical and horizontal motion generally reflect those in cumulative population data. From the distributions identified in Figure 3D, we generated parametric maps in which voxels are color coded according to orientation preference and superimposed on the fluorescence however image of the tectal neuropil (Figures 3E and 3F). These maps, which allow examination of functional architecture in individual larvae, reveal that in all subjects, orientation-selective inputs are broadly distributed across SFGS

and that voxels tend to cluster according to orientation preference. What is evident from the two examples of separate larvae is that within the orientation-selective domain, the organization of the two subtypes can be variable across subjects. The same orientation-selective inputs, with similar tectal distributions, were identified using the OSI metric (Figure S3). This figure also shows examples of single orientation-selective RGCs expressing SyGCaMP3 that are selective for either horizontal or vertical motion. The functional parametric maps of individual larvae shown in Figures 2 and 3 suggest regional differences in the distribution of direction- and orientation-selective inputs to the zebrafish tectum. To examine in more detail the spatial organization of direction- and orientation-selective responses in SFGS, we spatially coregistered data from all larvae to create single composite maps for each parameter (see Supplemental Experimental Procedures).

Gabbe et al.60 found age related differences

in body weight, hip flexor flexibility, and ankle plantarflexor flexibility, but could not explain the connections between these age-related differences and age-related differences in hamstring strain injury rate. Different hamstring injury rates in athletes of different races have been repeatedly reported in the literature. Verrall et al.2 found that Australian football players who were of aboriginal descent had a significantly higher risk of hamstring injuries in comparison to players of other races. Woods et al.8 reported ERK pathway inhibitors that English professional soccer players of African descent have a significantly higher risk of hamstring strain injury in comparison to players of other races. Brooks et al.6 noticed that, although not statistically significant, the incidence of hamstring strain injury among African CHIR-99021 cell line and Caribbean

descents was almost four times that of Caucasian players. These results suggest that individuals of difference races may have different muscle fiber compositions. Ama et al.85 demonstrated that individuals of African descent have more fast fibers than Caucasians. As previously mentioned, athletes who have more fast fibers may be prone to muscle strain injury. Woods et al.8 also argued that the increased pelvis anterior tilt of African descents might be another explanation of their elevated hamstring strain injury risk. However, a study by Mosner et al.86 found no difference in actual pelvis anterior tilt between African and Caucasian individuals. Many studies have demonstrated that a history of hamstring strain injury is a significant risk factor for the recurrence of the injury.2, 4,

6, 60, 67, 87, 88 and 89 Engebretsen et al.58 suggested that previous injury was the only significant risk factor for new hamstring strain injury for a group of Norwegian soccer players. Based on an animal experiment, Nikolaou et al.37 suggested that scarring and fibrosis seen in the muscle 7 days after initial strain injury may explain the elevated risk of the injury. As previously mentioned, Brockett et al.51 demonstrated that the legs Mephenoxalone with hamstring muscle strain injury histories had a significantly greater knee flexion angle for the maximum knee flexion torque in comparison to the legs without hamstring muscle strain injury histories. This indicates a possibility that previous strain injury resulted in shortened optimum lengths of hamstring muscles and thus increased the risk for injury. However, a recent prospective study by Fousekis et al.64 reported that previous hamstring injury significantly decreased the odds of injury. A possible explanation for this result is that rehabilitation programs might have eliminated some risk factors or reduced the effects of risk factors for the subjects involved.

We further challenged a valence-specific role for αβ neuron subsets

using additional genetic approaches. We first confirmed that αβs neurons are required for both appetitive and aversive memory retrieval using NP5286, another LGK-974 in vivo GAL4 line with strong expression in αβs neurons and weaker expression in αβp neurons (Figures 3A and S1F; Tanaka et al., 2008). Appetitive and aversive memory performance of NP5286;shits1 flies was statistically different to that of shits1 and GAL4 control flies ( Figures 3E and 3F). No statistical differences were apparent when experiments were performed at permissive 23°C ( Figure S3) and the NP5286;shits1 flies exhibit normal olfactory acuity at the restrictive temperature ( Table S1). We next challenged an appetitive memory-specific role for αβc neurons using an intersectional genetic strategy. Combining a ChaGAL80 transgene with c739 removes expression in the αβs and αβp neurons from the c739-labeled αβ population and leaves robust expression in αβc neurons (Figures 3B and S1G). We again trained flies at the permissive Metabolisms tumor temperature and blocked αβc during retrieval. Similar to the analysis with NP7175;shits1

flies, appetitive memory performance of c739;ChaGAL80/shits1 flies was impaired, being statistically different to the relevant control groups ( Figure 3E). Moreover, the c739 disruptive effect on aversive memory was abolished with ChaGAL80, consistent with removal of αβs expression from c739 ( Figure 3F). Control experiments at 23°C did not reveal significant differences between the relevant groups ( Figure S3). A role for αβc in memory consolidation has been reported (Huang et al., 2012). Blocking NP6024-labeled αβc neurons for several hours after training disrupted appetitive and aversive memory consolidation, whereas blocking NP7175-labeled neurons only impaired aversive memory consolidation

(Huang et al., 2012). Although others defined the αβ neurons labeled in NP6024 as inner and outer αβc neurons (Tanaka et al., 2008 and Huang et al., 2012), our anatomical analysis revealed that outer αβc neurons occupy the area of the vertical MB lobe that is anatomically indistinguishable from that containing oxyclozanide the αβs neurons (Figures 3A–3D). Furthermore, blocking output during retrieval in NP6024;shits1 flies significantly impaired both appetitive and aversive memory ( Figures 3E and 3F), consistent with NP6024 expressing in αβc and αβs neurons. Control experiments at the permissive temperature did not reveal significant differences between the relevant groups ( Figure S3) and the NP6024;shits1 flies exhibit normal olfactory acuity ( Table S1). Perhaps more importantly, a consolidation effect cannot account for our appetitive memory retrieval-specific function because the retrieval role for αβc is not time dependent.