, 2006). In our study, we found that elevated mTOR signaling in POMC neurons increased KATP current, and this heightened KATP channel activity silenced POMC neurons and reduced leptin-stimulated α-MSH secretion; Pomc-cre;Tsc1-f/f mice also exhibited a hyperphagic obese phenotype ( Figures 4 and 5). Selleckchem Rapamycin Since increasing PIP3 level on the plasma membrane activates PI3K, a canonical activator of mTOR ( Wullschleger et al., 2006), it is expected that deleting PTEN in POMC neurons may also activate mTOR, and this elevated mTOR activity could activate KATP channels, as we have shown in our study. Whereas deleting PTEN increases plasma PIP3 thereby prolonging

the opening time of KATP channels ( Plum et al., 2006), the elevated mTOR signaling in POMC neurons lacking TSC1 likely causes an ATM/ATR inhibitor increase in KATP channel density, because the maximum KATP current level is doubled in the presence of diazoxide ( Figure 4I). Since PIP3 and diazoxide

share the same common mechanism for KATP channel activation due to increased open time ( Koster et al., 1999), the maximum KATP current in POMC neurons without TSC1 should remain unchanged if activating mTOR were to increase KATP current by generating PIP3. It is of interest to note that deleting PTEN in POMC neurons also results in hypertrophic soma as in POMC neurons with an elevated mTOR activity ( Mori et al., 2009). It thus seems likely that activation of the PI3K pathway will have effects similar to those caused by elevating mTOR activity, likely an increase of KATP channel density, in addition to an increase of channel open time due to an increase of phosphoinositides old such as PIP3. Rapamycin has been found to affect the expression of Kv1.1 and Kv4.2 in dendrites of hippocampal

neurons (Lee et al., 2011; Raab-Graham et al., 2006). Here we provide another example of how mTOR regulates neuronal activity by controlling ion channel density. Under physiological conditions, the ion channel density in neurons is tightly regulated (Ma and Jan, 2002). For example, when Parton et al. (2007) expressed in transgenic mice a mutant form of Kir6.2 under the POMC promoter, POMC neurons in these transgenic mice nonetheless exhibit normal levels of KATP channel density (Parton et al., 2007). Ion channel density may be controlled at several different levels including transcription, translation, trafficking, and quality control of the endoplasmic reticulum (ER) (Ma et al., 2001). We found that POMC neurons from old mice express the transcripts for KATP channel subunits Kir6.2 and SUR1, for the most common KATP channel composition in neurons (van den Top et al., 2007). As functional KATP requires the coassembly of Kir6.2 and SUR1 (Schwappach et al.

This suggests that the piriform cortical ensembles completed the slightly degraded input, and responded as if the entire odor object was present. As more components were removed or novel contaminants added, piriform cortical ensembles decorrelated the mixtures even more strongly than the olfactory bulb (Barnes et al., 2008). Behavioral discrimination performance in a two-alternative choice task mirrored the cortical ensemble decorrelation—mixtures not decorrelated by the cortex were difficult for the animals to discriminate (Barnes et al., this website 2008). These results suggest that, as originally hypothesized (Haberly, 2001), the piriform

cortex can perform pattern completion which contributes to perceptual stability. Interestingly, new data suggest that the boundary between cortical

pattern completion and separation is experience dependent. Cortical pattern completion can be enhanced in tasks requiring odor generalization and pattern separation can be enhanced in tasks requiring fine odor acuity (J. Chapuis and D.A. Wilson, 2010, Soc. Neurosci., abstract). These changes in olfactory cortical processing lead to changes in perceptual acuity in both rodents (J. Chapuis and D.A. Wilson, 2010, Soc. Neurosci., abstract; Chen et al., 2011 and Fletcher and Wilson, 2002) and humans (Li et al., 2008 and Li et al., 2006). An autoexcitatory cortical network such as the piriform www.selleckchem.com/products/z-vad-fmk.html cortex Mephenoxalone is susceptible to run-away excitation and seizure activity. Thus, synaptic inhibition plays an important role in maintaining circuit function within manageable extremes. However, inhibition can also play important roles

in shaping receptive fields of individual neurons, and imposing temporal structure in pyramidal spike trains and circuit oscillations. These factors have been assumed in past cortical models but were never fully developed (Haberly, 2001). We now know that there are a large variety of inhibitory interneurons in piriform cortex falling into perhaps five different classes based on morphology, location, and physiological properties (Suzuki and Bekkers, 2010a, Suzuki and Bekkers, 2010b, Young and Sun, 2009 and Zhang et al., 2006), and these new data suggest unique roles for different cell classes. Interneurons have somata in all three layers of the piriform, and connections that can either remain within the same layer or spread to other layers. In addition to interlaminar connections, the connectivity and function of inhibitory interneurons may also vary over the anterior-posterior extent of the piriform cortex. For example, pyramidal cells in anterior piriform cortex appear to be under stronger inhibitory control from interneurons more caudal to them than interneurons more rostral (Luna and Pettit, 2010).

0 (SPSS IBM, Armonk, NY, USA), except the limits of agreement analysis which was conducted using MS Excel 2010 (Microsoft, London, UK). The self-selected pacing strategy inherent with the t-6MWT produced significantly different speeds (Fig. 1A) during the protocol (f   = 12.26, p   < 0.001). This was characterised by a conservatively paced minute 1 which was significantly slower than any other time during the protocol (p   < 0.05). Speed between minutes 2 and 4 was unchanged (p   > 0.05), but increased at minute 5 (p   = 0.004). Whilst there was not a continued increase to the end of the test, the speed at minute 6 was significantly selleck chemical greater than minutes 1–4

(p   < 0.05). The V˙O2 response ( Fig. 1B) was significantly increased during the protocol (f

  = 94.03, p   < 0.001) and correlated to the speed profile (r   = 0.779, p   < 0.001). Significant increases occurred at minutes 2 (p   < 0.001), 5 (p   = 0.002), and 6 (p   = 0.030). http://www.selleckchem.com/products/XL184.html Conversely BR ( Fig. 1C) significantly declined (f   = 91.14, p   < 0.001) and was negatively correlated with both V˙O2 (r = −0.686, p < 0.001) and self-selected walk speed (r = −0.411, p < 0.001). Decreases were observed each minute (p < 0.01) except between minutes 3 and 4 (p > 0.05). The mean ± SD 6MWD and 6MWW achieved during the t-6MWT was reported at 749.53 ± 94.30 m and 49,714.21 ± 9173.0 kg/m, respectively. For the most part 6MWW was more strongly related to the demographic and resting pulmonary function variables than 6MWD (Table 2). Correlation coefficients for V˙O2 compared to 6MWD and 6MWW were also assessed. The

6MWD portrayed a moderate correlation (r   = 0.656, p   < 0.001) with minute-to-minute V˙O2 ( Fig. 2A), but a slightly stronger relationship ( Fig. 2B) was observed between 6MWW and V˙O2 (r = 0.703, p < 0.001). Moves detected by the MWK were compared to 6MWD (Fig. 3A) and 6MWW (Fig. 3B), and a significant correlation was established with 6MWD (r   = −0.847, p   < 0.001) but not with 6MWW (r   = 0.337, p   = 0.220). Mean V˙O2 showed a moderate relationship (r = −0.753, p = 0.001) to moves accrued over the t-6MWT. The MWKEE demonstrated a notable relationship to 6MWW (r = 0.938, p < 0.001) ( Fig. 3D), but this was not the case between Idoxuridine MWKEE and 6MWD (r = 0.477, p = 0.720) ( Fig. 3C). The duration of time spent at different exercise intensities (Fig. 4) were significantly different between methods of calculation for moderate (χ2 = 15.25, p < 0.001) and vigorous intensities (χ2 = 19.63, p < 0.001), but not light intensity (χ2 = 4.00, p = 0.135). The MWK was not significantly different to measured (p > 0.05), but this was not the case for the standard method, which underestimated time durations at moderate intensity (p < 0.01) and over estimated at vigorous intensity (p < 0.01). Despite a strong correlation between MWKEE and gas analysis EE (r = 0.949, p < 0.001), the total EE was significantly different between the two methods (t = −6.76, p < 0.001).

Finally, genetic rescue experiments reveal that Cdh11 is one of the targets of the Bhlhb5/Prdm8 repressor complex whose upregulation in the absence of the Bhlhb5/Prdm8 repressor complex contributes to the abnormal phenotype observed in Bhlhb5 and Prdm8 mutant

mice. Taken together, these findings suggest that Bhlhb5 and Prdm8 are obligate components of a transcriptional repressor complex. Bhlhb5 binds to specific DNA within the regulatory regions of its target genes and recruits Prdm8 to mediate repression of the transcription of these targets. In the absence of Bhlhb5 or Prdm8 the gene targets are upregulated resulting in abnormal development of specific neural circuits. Since Bhlhb5 and Prdm8 belong to conserved gene families,

our finding Proteases inhibitor that these two factors specifically interact raises the question as to whether the functional association between bHLH factors and Prdm-related proteins is a more general occurrence. Phylogenetic analysis of SET-domain containing proteins reveals that Prdm8 is most closely related to Prdm13, and recent studies show that Prdm8 and Prdm13 are expressed in nonoverlapping patterns in the developing nervous system (Fumasoni et al., 2007 and Kinameri et al., 2008). However, we unexpectedly discovered that the gene with which Prdm8 shares the highest degree of similarity (as revealed using a blastn algorithm comparing murine genes) is not a member of the Prdm family, but rather the zinc-finger protein, Zfp488. This Selleck Dactolisib gene shares 82% similarity with Prdm8 over 80 amino acids in the C terminus (Figure 8C), suggesting that Prdm8 and Zfp488 may share a common ancestor. The idea that Prdm8 and Zfp488 are ancestrally related proteins is noteworthy

because Zfp488 was recently shown to interact physically and functionally with one of Bhlhb5′s closest relatives, Olig2 (Wang et al., 2006). This intriguing connection secondly suggests the possibility that the interaction between bHLH transcription factors and Prdm-related proteins is a general mechanism for the regulation of gene expression during neural development (Figure 8B). In this regard, it is notable that both Prdm13 and Olig3 are expressed in Class A progenitors in the dorsal spinal cord, consistent with the idea that these two factors may also couple selectively to mediate transcriptional repression in these cells (Kinameri et al., 2008 and Muller et al., 2005). We provide phenotypic and mechanistic evidence that Bhlhb5 and Prdm8 are obligate partners for certain aspects of development. However, it is likely that Prdm8 functions without Bhlhb5 in some contexts. This prediction is based on the observation that, while Prdm8 shows significant overlap with the Bhlhb5 expression domain, there are regions of the nervous system that express Prdm8 but not Bhlhb5.

This modification arose from several practical observations. First, nearby voxels share nonbiological signal (causing increased rs-fcMRI correlation), a result of unavoidable steps in data processing (e.g., reslicing, blurring). Second, short-distance relationships are especially susceptible to spurious augmentation by subject motion (Power et al., 2011). Third, as will be seen shortly, voxelwise graphs are dominated at higher thresholds by short-distance relationships, which are logically partially artificial based

on the above considerations. buy MK0683 Modified voxelwise networks are presented in which all ties terminating within 20 mm of a source node are excluded, though other distances (e.g., 15 mm and 25 mm) were also tested, with similar results (data not shown). The two standard methods of graph formation were parcel-based and voxel-based. The parcel-based graph was formed Selleck PD-L1 inhibitor using the 90-parcel AAL atlas (Tzourio-Mazoyer et al., 2002), a popular method of graph formation. This atlas divides the cortex and subcortical structures into parcels based upon anatomical landmarks. The voxel-based graph was defined using all voxels within the AAL atlas (n = 40,100), and the modified voxelwise graph was also defined using these voxels. Subgraphs were determined over a range of thresholds for each graph using one of the best-performing subgraph detection algorithms currently available

(Infomap) (Fortunato, 2010 and Rosvall and Bergstrom, 2008). This algorithm uses the map equation to minimize information theoretic descriptions Resminostat of random walks on

the graph (essentially assigning zip codes to subgraphs to shorten addresses of individual nodes). Other algorithms were tested and yielded similar results (Figure S2). Figure 1 illustrates our methodology and highlights several important results. The first panel depicts the areal graph in a spring embedded layout and maps subgraphs onto nodes using colors, visibly demonstrating the basis for subgraphs. In spring embedded layouts, ties act as springs to position nodes in space such that well-connected groups of nodes are pulled together, providing an intuitive and informative picture of the graph. The second panel shows the subgraph assignments of the areal network in both cohorts over a range of thresholds (each chart consists of 9 columns of 264 color entries). ROIs are ordered identically for both cohorts, and the patterns of subgraph assignment across cohorts are in good agreement. The standard graph theoretic measure of similarity between two sets of node assignments is normalized mutual information (NMI), which measures how much information one set of assignments provides about another set of assignments. Values of 1 indicate identical assignments, and values of 0 indicate that no information is gained about the second set of assignments by knowing the first. Between cohorts, NMI ranges from 0.86 to 0.

The identification of parasite taxa was performed using specialized literature and consulting paratypes. Values of the relative condition factor were obtained for all individual as described by Le Cren (1951). With the logarithms of the values of standard length

(Ls) and total weight (Wt) of each individual host, the curve was adjusted for Wt/Ls (Wt = a·Ltb) and the values of the regression coefficients a and b were estimated. The values of a and b were used for estimating values theoretically predicted of body weight (We) by using the equation: We = a·Ltb. Then the relative condition factor (Kn) was calculated, which corresponds to the ratio between the observed weight and the theoretically expected weight for a given length (Kn = Wt/We). The non-parametric test Kruskal–Wallis (H) was used to test differences of the mean Kn between the environments in the floodplain of the Upper Paraná River, www.selleckchem.com/products/LBH-589.html considering that different biotopes can influence the Kn. To assess the relationship between infracommunities and infrapopulations of parasites with the relative condition factor, the nonparametric Spearman’s rank correlation coefficient (rs) was applied for the variables Kn of each parasitized fish × total number of species in the infracommunities, Kn of each parasitized fish × total number of individuals in the infracommunities and Kn of each fish × abundance of each parasite species. Analyses in

the infrapopulations were applied to species with prevalence higher than 10%, as suggested by Bush et al. (1990). The Mann–Whitney’s U-test with correction selleck screening library for ties – Z(U) – was used to test differences between the mean Kn of males and females and parasitized and non-parasitized fish for each species ( Zar, Ribonucleotide reductase 1996). The level of significance

adopted was p < 0.05. Fifty-eight taxa of metazoan ecto and endoparasites were identified. Leporinus lacustris harbored 31 species, Leporinus friderici 32, Leporinus obtusidens 28 and Leporinus elongatus 25 species. Taxa recorded in the four host species, sites of infection/infestation and parasitism indicators are presented in Table 1. The mean values of the host’s Kn for total sample are presented in Table 2, where the means for males and females are also shown. The Kn did not differ significantly between males and females (L. lacustris: Z(U) = 1.776, p = 0.075–L. friderici: Z(U) = 1.849, p = 0.064–L. obtusidens: Z(U) = 0.693, p = 0.486–L. elongatus: Z(U) = 0.477, p = 0.632). It also did not differ between hosts collected in three types of environments (L. lacustris: H = 0.359, p = 0.825–L. friderici: H = 1.410, p = 0.493–L. obtusidens: H = 1.162, p = 0.559–L. elongatus: H = 0.812, p = 0.661). Thus, all fish of each species were treated as one data set. Among the analyzed specimens of L. lacustris, 4 and 45 were unparasitized by ecto and endoparasites, respectively. For L. friderici these numbers were 4 and 27, for L. obtusidens 4 and 17, and for L. elongatus 2 and 9, respectively.

This perspective is astonishingly naive. Even among the most impressive reports of axonal growth to date, the overall restitution of axon number is far below normal innervation density. Extensive restoration of function may require restitution of neural circuitry to pre-lesion patterns

that, during development, formed as a result of a precise orchestration of genetic and epigenetic events sequentially over time. This collective set of developmental events included both intracellular mechanisms in the neuron and environmental HA-1077 mouse expression of diffusible guidance cues, extracellular matrix molecules and cell adhesion molecules in precise temporal and spatial gradients. Moreover, remyelination of every new axon segment may be required to overcome conduction block. This set of restorative events is unlikely to occur after adult injury. Accordingly, the extent to which nondirected or partially directed growth can be functionally beneficial, as opposed to deleterious (causing spasticity or cause pain),

remains to be determined. We have only recently reached the point that this question can even be addressed because, finally, there are manipulations that produce at least some growth past the lesion. Directed rehabilitation, trophic gradients and other means may be required to shape the nature of circuit reformation, but even under these circumstances, will the number, topography, and remyelination of newly growing axons be sufficient to improve function? Moreover, we must also

ask whether our most commonly used functional measures are relevant to humans. For example, is restoration Fossariinae of selleck chemicals llc walking ability in a quadrupedal rodent relevant to the bipedal locomotion of humans that requires fine control of posture and balance? Nonetheless, partial improvements in behavior (often optimistically referred to as “functional recovery” in the literature) can be meaningful and informative regarding cellular and systems-level mechanisms that are required to improve function. Screening tools such as the Basso-Beattie-Bresnahan (BBB) scale (Basso et al., 1995) provide a convenient starting point, but quantifiable ordinate measures that are directly related to particular axon systems are needed to definitively relate axon growth with recovery. The requirement that experiments pass the criterion of demonstrating “functional benefit” to be considered of major importance in the spinal cord injury field should be soundly rejected by investigators, reviewers, and journal editors. We remain at a stage of spinal cord injury research in which discovery of fundamental mechanisms contributing to new axonal growth is critical: from new mechanistic discoveries that lead to significant axonal sprouting and regeneration, we will sequentially amplify the number of growing axons, the distance over which they grow, and their guidance to and connection with appropriate targets.

These latter findings raise the intriguing idea that cue-evoked states of gustatory expectation may generate a “preplay” of early information coding in response to unexpected taste. Well-designed control experiments helped rule out the possibility that cue-evoked responses in GC could have arisen from expectation-related differences in motor activity, including lever pressing, mouth movements, or selleck products other oromotor reactions. To the extent that the prestimulus, cue-related effects in GC are in fact anticipatory, it reasonably follows that these responses might be under top-down control. In order to test this hypothesis, the investigators performed dual recordings from GC and from the basolateral

amygdala (BLA), a region that has been implicated in network processing of taste coding (Grossman et al., 2008) and anticipatory states (Roesch et al., 2010), and sends direct projections to rodent GC (Saper, Linsitinib 1982). Like GC, the BLA responded to the auditory cues, but even more quickly, such that the average latency of cue-induced activity in BLA was on average 16 ms shorter than that of GC, a significant effect. These data, along with the finding of a cue-dependent strengthening of cross-correlation values between

BLA and GC, are consistent with a modulatory influence of BLA on anticipatory activity in GC. Finally, to confirm whether BLA played a causal role in GC response dynamics, cue-evoked activity was examined before and after inactivation of the BLA, through local bilateral injection of NBQX, an AMPA receptor antagonist. This manipulation impressively abolished the cue-evoked

activity in GC, highlighting the direct involvement check of BLA in establishing gustatory states of cortical expectation. Together these findings extend the traditional role of BLA in enriching sensory codes with emotional value. The findings presented here mark an important first step in understanding how expectation influences circuit activity in rodent GC, and add important information to the small but growing body of work exploring the neurocognitive interactions among attention, expectation, and chemosensory processing (Kerfoot et al., 2007, Nitschke et al., 2006, Saddoris et al., 2009, Stapleton et al., 2007, Veldhuizen et al., 2007, Veldhuizen et al., 2011, Zelano et al., 2005 and Zelano et al., 2011). The intriguing demonstration of gustatory information playback in GC during taste expectation raises an important question: what exactly is being played back prior to taste delivery? In the experimental design, the cue signaled to the rat that taste was imminent, but contained no information about stimulus identity or valence. Therefore the anticipatory activity in GC cannot be said to be playing back sensory-specific information about a particular stimulus.

A

null Selleck Quisinostat allele jkk-1(km2) ( Kawasaki et al., 1999) suppressed the arl-8 phenotype to the same degree as wy733 and wy735 ( Figure 1D), indicating that loss of JKK-1 activity suppresses the arl-8 phenotype. We used the jkk-1(km2) allele for our subsequent analyses. We quantified the suppression of arl-8 by jkk-1. Compared to the arl-8 single mutants, SNB-1::YFP is redistributed into more distal axonal regions in arl-8; jkk-1 double mutants ( Figures 1I and 1J). The size of the proximal SNB-1::YFP puncta (0–25 μm from commissure) is also significantly reduced in the double mutants ( Figure 1J). Furthermore, while the arl-8 mutants exhibited reduced number of presynaptic SNB-1::YFP puncta, this defect is abolished in the arl-8; jkk-1 double mutants ( Figure 1K). Similar results were obtained with two additional SV proteins, RAB-3 ( Figures S2A–S2D) and SNG-1/synaptogyrin (data not shown). Therefore, jkk-1 mutations partially and strongly suppressed multiple aspects of the arl-8 mutant phenotype in DA9. MAP kinases (MAPKs) act in cascades in which each MAPK is activated via phosphorylation by a MAPK kinase (MAPKK), which is in turn activated by a MAPKK kinase (MAPKKK) (Davis, 2000). JKK-1 was shown to be a specific upstream activator of c-Jun N-terminal kinase (JNK)-1, a homolog of mammalian JNK3 (Kawasaki et al.,

1999). A null mutation in jnk-1, gk7, caused the same degree of suppression of the arl-8 phenotype as jkk-1(km2) ( Figures 1E and 1I–1K). Moreover, the degree learn more of suppression in arl-8, jnk-1; jkk-1 triple mutants is indistinguishable

from that in either double mutants ( Figures 1F and 1I–1K), indicating that jkk-1 and jnk-1 function in the same pathway. jkk-1 and jnk-1 mutants were previously shown to partially mislocalize SNB-1::GFP to the dendrite in the DD motoneurons ( Byrd et al., 2001). We found that Urease the jkk-1 and jnk-1 single mutants appeared grossly normal in SV protein localization in DA9 ( Figures 1G and 1H) and did not show mislocalization of SV proteins to the DA9 dendrite (data not shown). However, these mutants did exhibit subtle but significant decreases in SNB-1::YFP puncta size ( Figure 1J) and increases in puncta number ( Figure 1K), suggesting that JNK also promotes SV clustering in wild-type animals. To determine whether JNK functionally interacts with arl-8 broadly in the C. elegans nervous system, we examined several other neuron types, including the cholinergic motoneuron DB7, the GABAergic DD motoneurons and the thermosensory neuron AFD, all of which have a proximal axonal region devoid of presynapses and form en passant presynapses in the distal axon ( Hallam and Jin, 1998; Klassen and Shen, 2007; Hellman and Shen, 2011).

We conclude that the ionotropic activity

of postsynaptic glutamate receptors, triggered by miniature events, is required for synapse growth. Because our results established that reduction of miniature neurotransmission inhibited synaptic development, we next investigated if increasing these events could also change synapse morphology. Complexin proteins bind to neuronal Selleckchem VX-770 SNARE complexes and regulate neurotransmitter release (Brose, 2008). Mutants of Drosophila complexin (cpx) have a dramatic increase in spontaneous synaptic vesicle release and have increased numbers of synaptic boutons ( Huntwork and Littleton, 2007). We hypothesized that these two phenotypes could be causally related through increased miniature NT. To test this idea, we first measured evoked and miniature NT in cpx null mutants. We found no change in the eEPSP integral ( Figures 4A, 4B, and 4H) in these mutants, although eEPSP amplitudes were reduced compared to controls ( Figure S5A), consistent Androgen Receptor Antagonist with previous studies (

Huntwork and Littleton, 2007 and Iyer et al., 2013). In contrast, cpx mutants had a dramatic 81-fold increase (p < 0.001) in miniature NT ( Figures 4A, 4B, and 4I). Expression of a complexin transgene (UAS-Cpx) in MNs rescued cpx mutants, restoring miniature NT to control levels ( Figures 4C and 4I). When we measured the terminal morphology of cpx mutants, we observed a 44% increase (p < 0.001) in terminal area ( Figures 4J, 4L, and 4M) accompanied by a 32% increase (p < 0.001) in typical bouton numbers but a 47% (p < 0.01) decrease in the number of small boutons ( Figures S5B and S5C). This lead to a 64% decrease (p < 0.001) of the bouton size index ( Figure 4K). As with neurotransmission, rescue of cpx mutants with transgenic complexin Adenylyl cyclase restored terminal area and the bouton size index ( Figures 4J, 4K, and 4N). Therefore, cpx mutants

have larger synaptic terminals with a decreased fraction of small boutons, the inverse of vglutMN and iGluRMUT mutant phenotypes. We next wished to determine if evoked NT contributed to cpx mutant terminal phenotypes. We first analyzed the cpx1257 mutant allele, which has normal eEPSP amplitudes and kinetics ( Iyer et al., 2013) ( Figure S5A) but has similarly increased miniature NT to cpx null alleles ( Figures 4B, 4D, and 4I). We found that cpx1257 mutants had increased terminal areas with a decreased bouton size index not significantly different from cpx null alleles ( Figures 4J, 4K, and 4O). This indicated that the aberrant terminal overgrowth of cpx mutants was not due to abnormal evoked release. As a second test, we expressed PLTXII in MNs of cpx null mutants. As expected, this strongly inhibited evoked NT without significantly altering miniature events ( Figures 4E, 4H, and 4I). When we measured the terminal morphology of these animals, we found no change compared to cpx mutants alone ( Figures 4J, 4K, and 4P).