Over the training period, both knockout and wild-type mice learned to locate the hidden platform but upon platform removal, Decitabine supplier Mbnl2 knockouts crossed over the location of the platform significantly fewer times than wild-type mice, indicating a deficit in spatial reference memory

( Figure 3C). Since Mbnl2 knockouts exhibited impaired spatial memory on a hippocampal-dependent task, we performed electrophysiological recordings on hippocampal slices to evaluate the effects of Mbnl2 loss on NMDA receptor (NMDAR)-mediated synaptic transmission and synaptic plasticity (long-term potentiation [LTP]). Input-output curves of the NMDAR-mediated component of the field excitatory postsynaptic potentials (EPSPs) indicated significant effects of stimulation intensity [F (7, 161) = 91.77, p < 0.0001] exhibiting a decreased response in Mbnl2 knockouts when compared to wild-type selective HDAC inhibitors controls [F (1, 161) = 21.94, p < 0.0001] ( Figures 3D and 3E). The decrease in the NMDAR component of synaptic transmission was not due to a loss of synaptic input since the presynaptic fiber volley amplitude

was similar across the two groups ( Figure 3F). Furthermore, the input-output curves of the slope of the synaptic responses were not significantly different between Mbnl2 wild-type and knockout mice (data not shown), indicating that the difference was specific for NMDAR function. For analysis of synaptic plasticity, LTP-inducing stimulation was delivered to the test pathway. Although pattern stimulation induced LTP in wild-type mice compared to the nontetanized path [F (1, 11) = 16.63, p < 0.001], this LTP induction was not observed in five out of six Mbnl2 knockout mice (wild-type, 134.6 ± 8.09; Mbnl2 knockout,

104.9 ± 5.96) ( Figure 3G). Thus, loss of Mbnl2 expression results in decreased synaptic NMDAR activity, impaired LTP, and learning and memory deficits. Another neurologic phenotype, which emerged with low penetrance (<10%) next in Mbnl2 homozygous knockout males prior to weaning, was extreme hyperactivity followed by tonic-clonic seizures and death within 24 hr. To determine whether Mbnl2 deficiency promoted seizure activity, we compared Mbnl2 wild-type with heterozygous and homozygous male knockouts for seizure susceptibility using the GABA antagonist pentylenetetrazole (PTZ). After intraperitoneal injection of PTZ, mice were evaluated for seizure activity using a modified Racine scale and by measuring the time to the initial appearance of abnormal behavior (latency time). Remarkably, a low PTZ dose (40 mg/kg) was sufficient to generate enhanced seizure incidence, including tonic-clonic and clonic seizures, even in Mbnl2 heterozygous knockout mice ( Figure 3H).

Evidence from human patient studies suggests that the functional differences of the dorsal and ventral pathways are better explained by vision-for-action and vision-for-perception, respectively (Goodale and Milner, 1992). In fact, V4 receives mixed magnocellular and parvocellular inputs originating from the lateral geniculate nucleus (Ferrera et al., 1994a), as well as input from MT (Maunsell and Van Essen, 1983; Ungerleider and Desimone, 1986). These connections make V4 an area that has rich access to motion information in the visual check details stimulus. Furthermore,

it has been shown that top-down signals to V4 also contain motion information (Ferrera et al., 1994b). As a result, V4 is well activated when monkeys are viewing moving stimuli (Tolias et al., 2001; Vanduffel et al., 2001). Our findings further suggest that the motion information is actively processed

in this area. Note that V4 is much larger than MT, so V4 may contain a comparable number of direction-selective neurons as area MT. This may raise the question, “Why would both dorsal and ventral pathways participate in motion processing?” A reasonable assumption is that the same motion information needs to be processed in different ways in order to serve different purposes. For example, motion perception requires integration of local motions, while distinguishing a moving object c-Met inhibitor from its background requires motion differentiation (Braddick 1993). We found that the motion-processing organization in V4 is different from that in MT. For example, many direction-preferring domains in V4 are scattered singulars, while direction preference maps in MT are more uniform (Malonek et al., 1994; Xu et al., 2004; Kaskan et al., 2010). The mean direction selectivity of neurons recorded in the V4 direction-preferring domains (mean DI = 0.63; this study) is lower than that found in area MT (mean DI = ∼1;

mafosfamide Albright et al., 1984). V4 neurons also tend to be more activated by moving lines than by moving random dots (Baker et al., 1981; Vanduffel et al., 2001). In addition, motion adaptation could induce direction selectivity of V4 neurons (Tolias et al., 2005). These data suggest that the direction-selective neurons in V4 have very different receptive field features than do MT neurons. These differences could give us hints on the functional roles of direction-selective neurons in the ventral pathway. It is also possible that perception of motion might be a distributed process that is not limited to the dorsal areas. This idea is supported by a recent finding that MT does not process global motion (Hedges et al., 2011). Motion information is useful for object identification. Relative motion between an object and its background is an important cue for figure-ground segregation, especially when other types of cues are weak or ambiguous (e.g., camouflaged insects).

The upper marker of the heavy exercise intensity domain is the maximal lactate steady state (MLSS, the highest metabolic rate at which exercise can be sustained without an accumulation of blood lactate33) or, more often in young people, the critical power (CP, the highest metabolic rate at which V˙O2 can be stabilised below peak V˙O236 and 37). Exercise above MLSS or CP but below

peak V˙O2 is in the very heavy exercise domain and exercise above peak V˙O2 is in the severe exercise domain.38 With young participants it has been noted that small selleck compound breath-to-breath variations are inherent to children’s response profiles.39 This reduces the confidence with which pV˙O2 kinetic responses can be estimated and confidence intervals are likely to be beyond acceptable limits unless sufficient identical transitions are aligned and averaged to improve the signal to

noise ratio.40 Rigorously determined and interpreted data from young people are available in the moderate, heavy and very heavy intensity exercise domains.41, 42 and 43 The pV˙O2 response to a step transition has three phases. At the onset there is an immediate increase in cardiac output which occurs prior to the arrival at the lungs of venous blood from the exercising muscles. This cardiodynamic phase (phase I) which, in children, lasts ∼15 s is independent of V˙O2 at the muscle (mV˙O2) and reflects an increase in pulmonary SCR7 datasheet blood flow with exercise. Phase II, the primary component, is a rapid exponential increase in pV˙O2 that arises with hypoxic and hypercapnic blood from the exercising muscles arriving at the lungs. Phase II kinetics are described by the time constant (τ  ) which is the time taken over to achieve 63% of the change in pV˙O2. In phases I and II ATP re-synthesis cannot be fully supported by oxidative phosphorylation and the additional energy requirements of the exercise are met from body oxygen stores, PCr and glycolysis. During moderate intensity exercise with children pV˙O2 reaches a steady state (phase III) within about 2 min. In the heavy intensity exercise domain, the primary phase II oxygen cost is similar to that observed during moderate

intensity exercise but the overall oxygen cost of exercise increases over time as a slow component of pV˙O2 is superimposed upon the primary component and the achievement of a steady state might be delayed by ∼10–15 min. 44 In adults, at exercise intensities above the MLSS or CP the slow component of pV˙O2 rises rapidly over time and eventually reaches peak V˙O2 but this phenomenon has not been observed in children. 37 and 45 The mechanisms underlying the pV˙O2 slow component remain speculative but it has been established that ∼86% have been accounted for at the contracting muscles.46 During exercise above the TLAC the pV˙O2 slow component is associated with a progressive recruitment of additional type II muscle fibres with the low efficiency contributing to the increased oxygen cost of exercise.

The precise mechanisms behind the generation of time fields and whether other structures organize according to the time kept in the hippocampus remain to be seen,

but Kraus et al. (2013) make it clear that time and place coexist in the hippocampus. D.C.R. is supported by the Marie Curie Foundation (GA-2011-301674). M.B.M. is supported by the Kavli Foundation and a Centre of Excellence grant from the Norwegian Research Council. “
“Spontaneous or endogenously driven neural activity has been a focus of investigation in electrophysiology Hydroxychloroquine mouse for many decades (Buzsáki, 2009). In recent years, researchers have focused on fluctuations in blood oxygenation level-dependent (BOLD) activity acquired during a “task-free” or “resting” state, as the spatiotemporal structure of these signals has proven richly informative about the functional organization of the human brain (Raichle, 2011). Resting-state dynamics are commonly characterized via “functional connectivity,” which describes the statistical dependence of activity

at different locations in the brain. Resting-state functional connectivity is often computed via a Pearson correlation of fMRI BOLD signal time series recorded from different Ulixertinib in vivo voxels. Despite the unconstrained mental state in resting-state fMRI experiments, patterns of functional connectivity across the brain are quite reproducible within individuals and across large cohorts of participants (Biswal et al., 2010). This observation suggests that functional connectivity may be shaped by the underlying anatomical connectivity. This notion has gained support from direct

comparisons of anatomical and functional connectivity from in the monkey (Vincent et al., 2007) and human (Honey et al., 2009) brain, as well as from interventional studies demonstrating changes in functional connectivity after manipulations of the anatomical substrate (Johnston et al., 2008). In addition, computational models combining cellular biophysics and networks of synaptic connections can generate realistic functional connectivity patterns (Deco et al., 2011). Despite the growing promise of BOLD functional connectivity, important questions remain concerning the optimal data acquisition and analysis methods (Cole et al., 2010) and the spatiotemporal scales at which dynamical correlations usefully indicate functional properties of the brain. Does functional connectivity recorded with fMRI (a slow and indirect neural observation) relate to functional connectivity recorded more directly with invasive electrophysiological methods? Does anatomical connectivity predict resting-state BOLD functional connectivity at spatial scales finer than a cubic millimeter? Can patterns of correlation in the BOLD signal reveal intra-areal functional topographies? In this issue of Neuron, Wang et al. (2013) make significant progress toward addressing these questions. Their focus is on connectivity within area 3b and area 1 of the squirrel monkey somatosensory cortex.

The conidial concentration was quantified using a hemacytometer according to Alves (1998). The conidia aqueous 0.1% Tween 80 suspension was adjusted to 108 conidia/mL. The mineral oil proportions used to prepare the formulations were adapted from Angelo et al. (2010). The formulations contained 10, 15, or 20% sterile mineral oil (Vetec Química GSK1120212 datasheet Fina Ltda., Rio de Janeiro, Brazil) and were prepared with the following proportions: (i) 89% of the aqueous suspension, 10% mineral oil and 1% Tween 80; (ii) 84% of the aqueous suspension, 15% mineral oil and 1% Tween 80; and (iii)

79% of the aqueous suspension, 20% of mineral oil and 1% Tween 80. Conidial viability was determined by plating an aliquot of the aqueous suspension and each oil formulation on PDA medium plus 0.05% chloramphenicol followed by incubation at 25 ± 1 °C. Conidial germination was observed after 24 h and 48 h (Alves, 1998). Three groups were formed in the bioassays with aqueous suspensions: a control group treated with sterile distilled water and 0.1% Tween 80, and two groups treated with M. anisopliae s.l. or B. bassiana suspensions. In the oil formulation bioassays, three groups were formed for each oil concentration (10,

15 or 20%): a control group, treated with sterile distilled water, 1% Tween 80 and the respective mineral oil concentration, and the two other oil based formulations of M. anisopliae s.l. or B. bassiana, Veliparib concentration with the appropriate proportions of water, mineral oil, and Tween 80. All bioassays were repeated twice. Twelve groups with 10 females of similar weight were formed. Each female was weighed, identified and submerged for 3 min in 1 mL of the test materials. Afterwards, the females were labeled, attached to Petri dishes Sitaxentan and incubated at 27 ± 1 °C and RH ≥80%. The egg mass laid by each female was weighed daily

and placed into individual test tubes. The eggs were then incubated at the same temperature and RH to allow the larvae to hatch. The following parameters were evaluated: hatching percentage; egg production index (EPI) (EPI = weight of egg mass/initial weigh of engorged female × 100) (Bennett, 1974); nutritional index (NI) (NI = weight of egg mass/initial weigh of engorged female − residual weight of engorged females × 100) (Bennett, 1974); and percentage of tick control (CP). The reproductive efficiency (RE) (RE = weight of egg mass/initial weigh of engorged female × hatching percentage) was used to calculate the CP (CP = mean RE of control group − mean RE of treated group/mean of control group × 100) (Drummond et al., 1971). Engorged females were held in Petri dishes and incubated at 27 ± 1 °C and RH ≥80% for oviposition. The eggs laid until the tenth day of oviposition were used in the bioassay with eggs and larvae. Egg aliquots of 50 mg were placed in test tubes sealed with cotton plugs. Each group was formed by eight test tubes.

Accurate determinations of Na+ channel distributions require unbiased matching of a wide range of AP properties (amplitude, rate of rise, site of initiation) in morphological realistic models. Using this approach, estimates in large cortical pyramidal neurons indicate AIS-to-soma Na+ channel ratios of ∼50-fold (Kole et al., 2008), whereas in electronically compact dentate granule cells a ∼5-fold difference seems to suffice (Schmidt-Hieber and Bischofberger, 2010). In addition to their high density, the properties of Na+ channels SP600125 in the AIS are specialized, presumably to facilitate AP initiation in the AIS. For example, the voltage dependence of both activation and inactivation of AIS

Na+ channels is hyperpolarized by ∼10mV compared to Na+ channels at the soma (Figure 2C) (Colbert and Pan, 2002, Hu et al., 2009 and Kole et al., 2008). This observation is consistent with subunit-specific differences in the voltage dependence of Na+ channels (Rush et al., 2005), providing further evidence that the primary Na+ channel subunit in the AIS is Nav1.6

(Hu et al., 2009, Lorincz and Nusser, 2010 and Royeck et al., 2008). AIS Na+ channels in dentate granule cells have also been shown to activate and inactivate approximately two times faster than those at the soma (Schmidt-Hieber and Bischofberger, 2010). This observation selleck chemicals has been used to explain how a low density of Na+ channels in the AIS of cortical pyramidal neurons could generate fast rising APs in the AIS (Fleidervish et al., 2010). Faster channel kinetics, however, means less charge influx per channel, leading to smaller AP amplitudes for a given Na+ channel density. Rapid Na+ channel kinetics alone, therefore, is unlikely to explain AP generation in the AIS, leading to the conclusion that a high density of Na+ channels is likely to be an absolute requirement. Another specialized property of Na+ channels is that they below can be activated at subthreshold potentials, as well as undergo incomplete inactivation, leading to generation of the so-called

persistent Na+ current (INaP) ( Taddese and Bean, 2002). Presumably due to the high density of Na+ channels in the AIS, INaP has been found to be greatest in the axon ( Astman et al., 2006 and Stuart and Sakmann, 1995), where it has a significant influence on AP threshold ( Kole and Stuart, 2008 and Royeck et al., 2008). Activation of INaP is also thought to be important for generation of the AP afterdepolarization, and as such plays a role in the generation of high-frequency AP bursts ( Azouz et al., 1996). Recent data in cortical pyramidal neurons indicates that AP bursts also require INaP activation at the first node of Ranvier ( Kole, 2011). Na+ channels, and especially the Nav1.6 isoform, can also undergo transient reactivation upon repolarization, leading to generation of a resurgent Na+ current (INaR).

In other words, it may be hypothesized that patients with elevated baseline neutrophils are completely protected from any grade of chemotherapy-induced neutropenia and have a poorer prognosis, independently from chemotherapy dosing. Recently, the result of a large, multicenter, randomized trial of flat dosing versus intrapatient

dose escalation of single-agent carboplatin as first-line chemotherapy for advanced ovarian cancer has been published [60]. A total of 964 patients were randomized. Dose escalation was achieved in 77% of patients who had ≥1 cycle. Intrapatient dose escalation of carboplatin based on nadir blood neutrophils or thrombocytes was feasible and http://www.selleckchem.com/mTOR.html safe. However, it provided no improvement in PFS or OS compared with flat dosing. Selleck INCB018424 In multivariate analysis, high baseline neutrophil counts had a significant adverse prognostic value whereas nadir neutrophils counts were not statistically significantly associated with outcome. The authors concluded that baseline neutrophils over-ride nadir counts in prognostic significance and questioned the use of dose escalation as a standard practice. The clinical relevance of tumor-infiltrating

neutrophils has recently begun to emerge. Direct associations between tumor-infiltrating neutrophils next and poor clinical outcome have now been described for

several types of human cancer. The prognostic role of tumor-infiltrating neutrophils, elevated blood neutrophils and elevated blood neutrophil/lymphocyte ratio has been associated with poor clinical outcome, most notably in renal cell carcinoma, melanoma, colorectal cancer, hepatocellular carcinoma, cholangiocarcinoma, glioblastoma, GIST, gastric, esophageal, lung, ovarian and head and neck cancer. A striking finding is the notion that high baseline neutrophil count in either tumor or blood, or both, was identified as a strong, independent risk factor for poor outcome and the negative prognostic impact of neutrophils was not eliminated by increasing the dose of cytokines, chemotherapy, or targeted therapy. For several cancers, patients benefit most from therapy if baseline neutrophil was low. Thus, baseline neutrophils over-ride nadir counts in prognostic significance. This has especially been shown in kidney cancer, colorectal cancer, non-small cell lung cancer, ovarian cancer and nasopharyngeal carcinoma.

Transit amplification was demonstrated in the developing brain by direct visualization of intermediate progenitors undergoing division over time (Noctor et al., 2004). A similar experiment would test our model in the adult hippocampus. Cell survival impacts any lineage as it accumulates, but unfortunately the number of cells undergoing apoptosis over time is difficult to quantify due to transient expression of apoptotic markers and rapid clearance of dead cells. While some

indirect information about cumulative cell death can be achieved through BrdU survival studies, the technique provides limited information about total populations of cells (Taupin, 2007). BrdU survival experiments are least informative about slowly dividing NSC and rapidly dividing IP populations since the former are poorly labeled by BrdU, while decreased label retention in the latter can reflect either cell death or multiple divisions (Dayer et al., 2003, Encinas et al., Enzalutamide nmr 2011, Mandyam et al., 2007 and Taupin, 2007). Hence, the contributions of NSC and IP cell death during lineage accumulation and in response to environmental manipulations remain to be determined. Our own cell survival studies of neurons, which are postmitotic and thus lend themselves to BrdU survival studies, did 3-deazaneplanocin A solubility dmso not detect an effect of social isolation on survival (Figures S4A and S4B). In EEE-treated mice, in addition to observing a well-established and robust increase

of proliferation (data not shown), we also detected increased neuronal survival suggesting that decreased neuronal death contributes to the lineage gains described in this study. Our inability to detect any relationship between apoptosis and lineage expansion, within or between any of our groups, suggests that apoptosis (Figure S4D and S4F), while impacting accumulation of the lineage, is unlikely to account for the differences that we observe. An expanding stem cell compartment could allow the brain to grow a NSC reservoir during

deprived conditions such as isolation stress. This brain adaptation would then allow an augmented neurogenic response through experience-directed fate specification when the environment next became richer. An analogous type of proliferative control was recently recapitulated in a more artificial, embryonic stem cell system where extrinsic stimulation and activation of signaling pathways favored differentiation, while depleting these signals favored self-renewal (Ying et al., 2008). The signals dictating changes in the fate of the NSC lineage remain to be determined. Of particular interest are the multiple observations that neural activity is positively linked to cell division in the adult dentate gyrus (Deisseroth et al., 2004 and Tozuka et al., 2005). Environmental enrichment was previously demonstrated to increase neuronal activity in the dentate gyrus (Tashiro et al., 2007) while social isolation was recently demonstrated to decrease it (Ibi et al., 2008).

Mice were allowed to recover for 2–4 weeks to allow for viral expression before electrophysiology and imaging were performed. See Supplemental Experimental Procedures for details. Standard artificial cerebrospinal fluid (ACSF) consisted of NaCl (125 mM), NaHCO3 (25 mM), KCl (2.5 mM), NaH2PO4 (1.25 mM), 3-MA molecular weight MgCl2 (1 mM), CaCl2 (2 mM), glucose (22.5 mM), Na-pyruvate (3 mM), ascorbate (1 mM). Sucrose-enriched modified dissection

ACSF contained NaCl (10 mM), NaH2PO4 (1.2 mM), KCl (2.5 mM), NaHCO3 (25 mM), glucose (25 mM), CaCl2 (0.5 mM), MgCl2 (7 mM), sucrose (190 mM), pyruvate (2 mM). The ACSF had a pH of 7.3, osmolarity of 305–320 mOsm, and was saturated with 95% O2 and 5% CO2. The intracellular solution contained KMeSO4 (135 mM) (for current-clamp recordings) or CsMeSO4 (135 mM) (for voltage-clamp recordings), KCl (5 mM), NaCl (2 mM), EGTA (0.2 mM), HEPES (10 mM), phosphocreatineNa2 (10 mM), MgATP (5 mM), Na2GTP (0.4 mM), Alexa Fluor 594 (0.1 mM), and Biocytin (0.2%). In a subset of experiments, the following drugs (Tocris) were used at the following concentrations via bath application (unless otherwise noted): SR95531 (2 μM), CGP55845 (1 μM), AM251 (2 μM), NBQX (10 μM), D-APV (100 μM), and LY 367385 (100 μM). RuBiGABA was obtained from Tocris

or Ascent and PSEM308 was a generous gift from Scott Sternson and used at a concentration of 5 μM and 3 μM, respectively. A vibrating microtome (Vibratome 1000 or Leica CP-690550 solubility dmso VT1200S) was used to obtain 400-μm-thick horizontal or transverse sections of brains from mice that were transcardially perfused with ice-cold dissection ACSF. Slices were allowed to recover for at least 30 min at 34°C and then Thalidomide stored at room temperature in a 50% dissection:

50% standard ACSF solution. Infrared- or fluorescence-guided whole-cell patch-clamp recordings were performed at 34°C in standard ACSF. Fire-polished borosilicate glass pipettes (Sutter) were used with tip resistances of 3.5–4.5 MΩ for somatic and 8–10 MΩ for dendritic recordings. A Multiclamp 700B Amplifier and pClamp 9 software (Axon Instruments) were used for data acquisition. The average series resistance for whole-cell voltage-clamp recordings was kept between 9–15 MΩ; 75%–80% of the resistance was compensated. Current-clamp recordings were obtained with access resistances of 10–20 MΩ for the soma and 10–40 MΩ for the dendrites, compensated in bridge mode. ITDP was induced by paired PP and SC electrical stimulation at a −20 ms interval (PP before SC) at 1 Hz for 90 s using focal glass pipette-stimulating electrodes coupled to constant current stimulators (WPI). Stimulus strengths were adjusted so that PP and SC PSPs were less than 50% of their maximal amplitude (typically <0.5 mV for PP and <5 mV for SC).

The expression of Shh and its receptor Boc by two complementary nonoverlapping populations CP-673451 of neurons during synaptogenesis suggests a mechanism for achieving specificity of circuitry, where the target cell expresses the ligand (Shh) and the presynaptic cell expresses the corresponding receptor (Boc) (Sanes and Yamagata, 2009 and Williams et al., 2010). To examine whether the Boc mutant animals have a similar cortical phenotype to ShhcKO mutants, we performed Golgi analysis on P20 brains of BocKO mice and wild-type littermate controls ( Figures 6A–6D). We observed significant reductions

in spine density, and growth and complexity of basal dendrites located in layer V neurons of BocKO animals ( Figures 6E and 6F), while there was no difference in branch growth and spine density in layer II/III ( Figures 6G and 6H). These findings suggest that the non-cell-autonomous decrease in dendritic growth of layer V neurons may be due to a loss of synaptic activity from presynaptic Boc expressing neurons ( McAllister et al., 1996). To test for the loss of presynaptic input from Boc expressing neurons we utilized in utero electroporation to introduce synaptophysin-GFP,

a marker for active presynaptic terminals ( Kelsch et al., 2008, Li and Murthy, 2001, Meyer and Smith, 2006 and Nakata et al., 1998), into lower layer II/III cortical neurons at embryonic day 14 (E14) ( Figures S6A–S6C). We targeted neurons in lower isothipendyl layer II/III because find more of the extensive number of these cells exhibiting LacZ reporter gene expression observed in the Boc gene trap reporter mice, and also because of the preference for neurons located in this layer to make synaptic connections onto layer V pyramidal neurons ( Anderson et al., 2010 and Petreanu

et al., 2007). In addition to synaptophysin-GFP, we coelectroporated a plasmid for a pCAG-mTdTom-2A-H2BGFP plasmid ( Trichas et al., 2008) that labels electroporated cells with a nuclear GFP, and a membrane TdTomato, in order to label axonal projections ( Figures 7A–7C). In BocKO mice we coelectroporated the synaptophysin-GFP along with the mTdTomato axonal marker. We also coelectroporated a short hairpin RNA targeted against Boc (Boc-shRNA) into the brains of wild-type non mutant mice that should have normal levels of Boc and Shh, except for the population of electroporated cells. When we compared the density of synaptophysin-GFP puncta located on the axons of Boc-shRNA expressing versus control-shRNA expressing cells, we found a significant reduction in the density of puncta on axons located in layer V ( Figures 7C–7F). Notably, this reduction was observed in both ipsilateral and contralateral layer V axons, while there was no significant difference in the puncta density in layers II/III ( Figure 7J). We found a similar pattern of reduced puncta when we compared BocKO and control animals ( Figure 7K).