” The three anatomical components that we will discuss in this review are the cerebellum, basal ganglia, and motor cortex (Figure 1). The review, not surprisingly, raises more questions than answers, and if anything should be considered a form of manifesto. The overall purpose is to call attention to the benefits of a comparative approach. First, we hope to show that explicit comparison of motor learning Alpelisib in vivo results across the various model systems currently under investigation can help support or refute viewpoints on the role of specific structures. Second, to inspire

experimental directions in any given model system that might otherwise not be considered. Finally, given that neurorehabilitation is predicated on motor learning (Krakauer, 2006), taking a closer look at how motor learning itself is accomplished after brain injury and disease in model systems may improve the way that we train patients to gain back their lost motor abilities. Motor learning is a blanket term for any practice-related change or improvement in motor performance for a defined

variable of interest. In this review, we will draw a broad distinction between two learning mechanisms—motor adaptation and skill learning. By motor adaptation we mean the fast changes that return behavior to baseline levels of performance in the setting of perturbations that induce systematic errors, for example, prism adaptation. By skill learning we mean the slower changes that lead to performance improvements

Phosphoprotein phosphatase that are better than click here baseline. Such behaviors include learning to ride a bicycle or to play the violin. In addition to these two kinds of motor learning, there is an intermediate category of learning that is more difficult to categorize but can be broadly captured by the idea of action selection. The whole field of reinforcement learning is predicated on the idea that particular actions come to be associated with successful goal completion. For example, completing a maze or learning to press a lever for food at particular intervals. The question is—is this motor skill learning? We would say no because the quality of the motor performance itself is not the metric of interest, instead the motor system is just used to read out whether operant learning has occurred. We will have more to say about this in the course of the review. For now we will restrict ourselves to the comment that it is of interest that many studies of skill have focused on sequence learning, in which the order in which actions must be performed is almost always emphasized over the quality of the execution of the actions themselves. There are clear preferences with regard to the kind of motor learning studied depending on the effector and model system used. For example, in the case of eye movements, the focus is mostly on adaptation (Schubert and Zee, 2010), indeed it is hard to imagine what a skilled eye movement would be.

Relatively rapid homeostatic scaling up of synapses can also be evoked acutely by blocking NMDAR-mediated suppression of local protein translation. This increase in AMPAR-mediated current results from activation of local protein synthesis and increased availability of AMPAR subunits (Ju et al., 2004, Sutton et al., 2004 and Sutton et al., 2006). We found that protein translation-dependent scaling is occluded in the absence of GluN2B and is not rescued in 2B→2A neurons, suggesting that NMDAR-mediated suppression of protein translation is subunit specific.

From this we infer that a dominant role for GluN2B-containing NMDARs during development is to maintain appropriate levels of protein translation in dendrites in order to regulate synapse excitability. Consistent with this, we observed increased levels selleck inhibitor of phosphorylated S6K in dendrites lacking GluN2B and increased surface expression of AMPAR

subunits GluA1 and GluA2 in dendrites of GluN2B null neurons (Hall et al., 2007). mRNAs encode GluA1 and GluA2 traffic to dendrites, where their translation is locally regulated. Interestingly, antagonism of GluN2B-containing NMDARs results in upregulation of synaptic protein http://www.selleckchem.com/products/ulixertinib-bvd-523-vrt752271.html translation in vivo through activation of the mTOR pathway (Li et al., 2010). This suggests that it is through regulation of local protein synthesis that GluN2B antagonists may exert their effects as strong antidepressants (Maeng et al., 2008, Preskorn et al., 2008 and Li et al., 2010). It will be critically important to determine the exact molecular mechanism by which NMDARs, and specifically GluN2B, regulate protein synthesis in neuronal dendrites and to Carnitine dehydrogenase identify the RNA messages involved. Our experiments suggest that the specificity of GluN2B function is mediated through its preferential association with CaMKII. Regulation of AMPAR-mediated currents at developing cortical synapses requires CaMKII function downstream of GluN2B, because expression of a subunit mutant unable to

interact with CaMKII is ineffective at rescuing GluN2B loss of function. Importantly, although we observed that levels of activated (phosphorylated) CaMKII are depressed in the absence of GluN2B signaling, we actually observed an increase in the protein expression levels of this kinase and a decrease in the levels of beta CaMKII (data not shown). Interestingly, bidirectional homeostatic synaptic plasticity has been shown to involve reciprocal regulation of alpha and beta CaMKII (Thiagarajan et al., 2002 and Groth et al., 2011). Thus, it will be important in future studies to determine the exact conditions and mechanisms by which these enzymes are regulated by GluN2B and GluN2A-mediated signaling.

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), selleck chemicals llc 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 Idelalisib molecular weight 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 PLEKHM2 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).

Blood DNAs (and a few rare cases, EBV-immortalized DNAs) from nearly 1000 families (of the 3000 planned) were sent to our group for processing and analysis. Approximately one-tenth of the families we analyzed are not yet officially in the SSC databases. DNA samples were shipped to NimbleGen’s Icelandic facility, where two-color hybridizations using a single reference male genome were performed. SSC samples were labeled with Cy3, and the reference was labeled with Cy5. Ninety-seven percent

of families passed gender and pedigree checks for all members selleckchem and are called “valid” herein. Those are the only families considered in this report. We define a trio as consisting of a mother, a father, and a child, either affected or unaffected. If each member of a trio has a hybridization that passes minimum quality thresholds (see Experimental Procedures), that trio and its associated hybridizations are called “high quality” (or “HQ”). Out of 1721 valid trios from 887 families, 1475 (86%) are HQ. For convenience, throughout this report we refer to the children with diagnosed ASDs as “probands” and to the children who do not have ASDs as “sibs.” For purposes of statistical evaluation, we establish the “HQ quads,” a subset of 510 HQ families with exactly one proband and one sib each. The composition of the children and families

for the various subpopulations under study is summarized in Table 1. There are roughly equal proportions of probands and sibs. The male-to-female ratio among the probands is 7:1, typical of high-functioning ASDs (Newschaffer et al., 2007). We mention here the observation (to be discussed later) that there are fewer male sibs than female sibs. G protein-coupled receptor kinase Hybridization Baf-A1 data underwent extensive

processing before determining segments of altered copy number (Experimental Procedures, Supplemental Experimental Procedures, and Figure 1). We extracted signal and noise parameters from each hybridization and used these for quality control and to model integer copy-number states (Figure 2). For partitioning the genome into intervals of constant copy number, we used KS segmentation (Grubor et al., 2009). We also employed a trio-based Hidden Markov Model (HMM) to build databases of high-confidence events and transmissions. High-confidence events in 1500 parents were used to compile a frequency table of copy-number variation for all probes. We searched for de novo events in the 1475 HQ trios, initially restricting evidence to autosomal probes that did not have known extra mappings to the human genome (hg18 build) outside the event region, and probes that were rarely polymorphic in the high-confidence parental database (i.e., present in no more than 5/1500 parents). We compiled those events with high statistical significance of being de novo (p value < 10−9), creating a “stringent” automated list of 70 de novo events (Table S1, “stringent”). Figure 3 illustrates the family probe ratio data for two typical de novo events, a duplication and a deletion.

While we are awaiting the results of multiple large-scale sequencing efforts, the field is poised to move

on to functional studies that will help understand the molecular underpinnings and neural substrates of this disorder in hopes of developing more effective interventions. “
“The assembly of a highly organized network of neuronal connections is a key developmental process and essential for all neural function, ranging from simple movement to complex cognitive processes. Research focused on the cellular strategies and molecular mechanisms that orchestrate VX-809 cell line neural network assembly led to the discovery of a wide variety of axon guidance molecules and receptors (Kolodkin and Tessier-Lavigne, 2010). Many guidance molecules are evolutionarily conserved and, based on their mode of action, are categorized into short- buy Alisertib or long-range guidance cues that influence growth cone steering in a positive (attractive) or negative (repulsive/inhibitory) manner. We now know that the activity of an individual guidance cue is not absolute, but instead interpreted by the neuronal growth cone in a context-dependent manner.

Important conceptual advances in deciphering the molecular language of axon guidance and network assembly include the discovery of hierarchies among guidance cues, the identification of molecular switches that when flipped turn an attractive cue into an inhibitory one (or vice versa), and the existence of diverse receptor complexes that facilitate cell-type-specific responses to a specific guidance cue. The discovery of general principles underlying

the wiring of the developing nervous system provides insight into the molecular logic that allows a relatively small set of guidance cues to initiate the Phosphatidylinositol diacylglycerol-lyase assembly of complex neural networks with myriad interconnected circuits. In this issue, Erskine et al. (2011) and Ruiz de Almodovar et al. (2011) now provide new evidence that a key angiogenic factor, VEGF-A, exhibits angiogenesis-independent chemoattractive effects on spinal commissural and retinal ganglion cell axons at the CNS midline. It is not by chance that analysis of nervous system midline development has been particularly successful in the discovery of guidance cues and the elucidation of axon pathfinding mechanisms. Axons extending toward the CNS midline during development must make an important decision: to cross and find a synaptic partner on the contralateral side of the nervous system (relative to their cell body) or not to cross and remain confined to the ipsilateral side. Extensive work in fruit flies, worms, fish, chicks, and mice has established that the midline is a rich source of chemoattractants and chemorepellents (Figure 1A) (Dickson and Zou, 2010). Vertebrate Netrin-1 is a robust chemoattractant for spinal commissural axons and is secreted by floor plate cells located at the ventral midline.

To address these issues, we first cultured NCAM−/− and NCAM+/+ commissural neurons and exposed them to control and gdnf application for quantification of Plexin-A1. We observed that NCAM loss prevented gndf-induced increase of Plexin-A1 levels ( Figures 7A and 7B). Second, the Plexin-A1/Nf160kD ratio in FP and PC domains was determined in E12.5 NCAM+/+ and NCAM−/− embryos. This analysis revealed that the ratio was significantly decreased in the PC domain of NCAM−/− sections, compared with the wild-type ones, although in the FP domain, Selleck C646 the ratio was not statistically significant ( Figures 7C and 7D). In this mouse line, a large amount of axons are still present in

the FP at E12.5 due to the genetic background (C57Black6) and have not yet Selleck LY2157299 experienced FP crossing. This might limit the possibility to detect moderate differences. Third, using the t-BOC reporter, we examined the calpain activity in NCAM+/+ and NCAM−/− commissural neurons. We found that the proportion of cells exhibiting high calpain activity (high t-BOC fluorescence) was significantly decreased by gdnf in the NCAM+/+, but

not NCAM−/−, condition ( Figures 7E and 7F). Similarly, application of a GFRα1 function-blocking antibody abolished the gdnf-induced decrease of high t-BOC-labeled neurons ( Figures 7G and 7H). Finally, calpain activity was measured in commissural tissue dissected from NCAM+/+ and NCAM−/− embryos, exposed to acute stimulation with gdnf and control. The analysis revealed that gdnf could decrease the endogenous calpain activity in NCAM+/+, but not in NCAM−/−, tissue ( Figure 7I). Altogether, these experiments provide evidence that NCAM and GFRα1 are required for the gdnf-induced regulation

of Plexin-A1 levels and calpain activity in spinal commissural neurons. We report here that a local source of gdnf in the FP acting through NCAM, but not RET, regulates the responsiveness of commissural axons to the midline repellent Sema3B. Gdnf makes an important contribution to this process but also acts with NrCAM in the FP to switch on the Sema3B repulsive signaling by inhibiting calpain1-mediated processing of the Sema3B coreceptor Plexin-A1. This mechanism prevents axons responding to Sema3B at the precrossing DOK2 stage, thus allowing them to enter the FP, and then switches on sensitivity to Sema3B at the postcrossing stage (Figure 8). The navigation of commissural axons across the FP has been shown to be a complex multistep process. Upon crossing, axons acquire responsiveness to local repellents to which they were not sensitive before crossing, thus gaining the information to move away (Evans and Bashaw, 2010). Sema3B was shown by previous work to be one such repulsive cue, expelling commissural axons that have crossed the midline (Zou et al., 2000; Nawabi et al., 2010).

The tests were conducted in the Department of Food Science and Human Nutrition’s sensory evaluation laboratory, which includes seven panelist booths, controlled lighting, and computers equipped with Sensory Information

Systems (SIMS version 6.0 software for data collection). For the triangle test, randomized three digit numbers were used to label X-ray treated and control almond and walnut samples. The control sample was the odd sample in half of the tests, and the treated sample was the odd sample in the other half. After being instructed on GSK1210151A in vitro how to evaluate the samples using an example tray and test demonstration, panelists were asked to pick the sample that differed from the other two in the almond check details triangle test. Panelists then were presented the walnut samples using the same protocol. If panelists could detect an overall difference between the control and test sample, an acceptance test would determine if this difference significantly affected consumer acceptability. Based on the triangle tests results, an acceptance test was run for the walnut control and irradiated samples to evaluate appearance, aroma, flavor, texture, and overall acceptability. The same dose that was used

for the difference test was applied to the nuts, and the acceptance test was conducted with 75 nut consumers. A nine point hedonic scale was used with 9 = “like extremely,” “8 = like very much,” “7 = like moderately,” 6 = like slightly, “5 = neither like nor dislike,” “4 = dislike slightly,” “3 = dislike moderately,” “2 = dislike very much,” “1 = dislike extremely. To investigate the fate of Salmonella during long-term storage, nuts were inoculated, irradiated, and held at 4 °C/70%RH. Almonds and walnuts were inoculated with SE PT30 and conditioned at 0.2 and 0.7 aw. Thereafter, the inoculated

samples were bagged inside the conditioning chamber to maintain the established aw and irradiated at doses of 1.13 (almond; SE PT30; 0.2 aw), 2.37 (walnut; SE PT30; 0.2 aw), 2.28 Adenosine (almond; SE PT30; 0.7 aw), 4.32 (walnut; SE PT30; 0.7 aw), 2.28 (almond; S. Tenn.; 0.7 aw), and 4.32 (walnut; S. Tenn.; 0.7 aw) kGy to achieve ~ 5 log reductions (not the presence/absence test) at the corresponding aw values. Therefore, plate counts were always positive and there was no need of enrichment process. The bags of SE PT30 inoculated nuts were opened and placed in a refrigerator at 4 °C/~70%RH for 7 days to equilibrate with the storage conditions. Thereafter, the bags were closed to prevent any further contamination and returned to storage for up to 120 days. In contrast, the bags of S. Tennessee-inoculated nuts were previously conditioned to 0.7 aw and consequently remained closed during storage. Three bags each of the irradiated and control nuts were randomly selected after 1, 30, 60, 90, and 120 days of storage and quantitatively examined for Salmonella as previously described.

, 2010). In other cases, bAPs prime the dendrite to produce synaptically evoked calcium spikes which mediate STDP-LTP (Zhou et al., 2005; Kampa et al., 2006) For more on dendritic excitability and STDP, see Sjöström et al. (2008). The decremental propagation of bAPs creates a profound spatial gradient of STDP in neurons. In L5 pyramidal cells in neocortex, brief pre- and postsynaptic spike trains evoke Hebbian STDP at proximal synapses (<100 μm from soma) but progressively less LTP at more distal synapses. The most distal synapses (>500 μm) show only anti-Hebbian LTD in

response to pre-leading-post pairing. Distal LTD can be converted to LTP by supplying sufficient dendritic depolarization to either enhance bAP propagation (Sjöström and Häusser, 2006) or convert the single bAP into a dendritic-somatic spike burst (Letzkus et al., 2006). Smaller L2/3 Entinostat in vitro pyramidal cells exhibit a similar Endocrinology antagonist trend in which distal synapses express less STDP and a broader LTD window than proximal synapses (Froemke et al., 2005). Thus, decremental bAP propagation creates distinct dendritic plasticity zones in which different rules for synapse modification exist ( Figure 4B; Kampa et al., 2007; Spruston, 2008). In general, the most proximal synapses experience the strongest bAPs and are expected to exhibit Hebbian STDP with minimal requirements for synaptic cooperativity

and firing rate. More distal synapses will exhibit LTD-biased Hebbian STDP ( Froemke et al., 2005) or anti-Hebbian LTD ( Sjöström and Häusser, 2006) and will require high firing rates or strong synaptic convergence for Hebbian STDP. These synapses can exhibit anti-Hebbian STDP, if post-leading-pre firing drives synaptically evoked calcium spikes ( Kampa et al., 2006; Letzkus et al.,

Fenbendazole 2006). Very distal synapses may be largely outside the influence of bAPs, so that STDP is absent and plasticity is induced by cooperative firing of neighboring inputs that evokes dendritic sodium or calcium spikes or regenerative NMDA spikes ( Golding et al., 2002; Gordon et al., 2006). The existence of different plasticity rules within dendritic regions may contribute to activity-dependent stabilization of different functional classes of synapses in these regions ( Froemke et al., 2005). Modulation of dendritic excitability will regulate both the shape of STDP rules and the spatial extent of dendritic plasticity zones, including increasing or decreasing the prevalence of STDP relative to local, associative forms of plasticity. Neuromodulation has robust effects on the spike timing dependence of plasticity. This includes gating of STDP, as in adult visual cortex slices, where exogenous activation of receptors coupled to adenylate cyclase (e.g., β-adrenergic receptors) and PLC (e.g., muscarinic acetylcholine receptors) are necessary for LTP and LTD, respectively, within Hebbian STDP (Seol et al., 2007).

Inhibition, which interacts intimately with excitation, slows down saturation and increases the input dynamic range. This

leads to a sharpening of selectivity of membrane potential responses. Our results demonstrate that inhibition plays an indispensable role in the generation of sharp OS in mouse simple cells. The broad inhibition revealed in these cells suggests that different cortical circuits combine excitation and inhibition in unique ways to produce OS. In this study, we focused on simple cells since they have been thought as the group of neurons in which OS first emerges. Different from cats, in the mouse V1, neurons exhibiting conventional simple-type receptive fields (RFs) are much more abundant in layer 2/3 than layer 4 (Liu et al., 2009). With loose-patch recordings, which detect spike signals signaling pathway from patched neurons without affecting their intracellular milieu, we BKM120 clinical trial first examined OS of simple cells in layer 2/3. The On/Off spatial RF was mapped to determine the cell type, and the relationship between the RF structure and OS was determined. As shown in Figure 1A, the example neuron displayed a typical simple-cell RF with spatially segregated On and Off subfields. When tested with drifting sinusoidal gratings, the cell responded maximally to vertically oriented gratings (Figure 1B). The cell’s preferred orientation is similar to

the orientation perpendicular to the RF axis, which is defined as the line connecting the centers of On and Off subfields (see Experimental Procedures). A summary

of 34 simple cells (Figure 1C) indicates a strong correlation between the preferred orientation and the RF axis, consistent with previous observations in the cat V1 (Lampl et al., 2001). According to this result, the preferred orientation of a simple cell can be predicted rather precisely from its On/Off RF structure. Alanine-glyoxylate transaminase By whole-cell current-clamp recording with a K+ gluconate-based intracellular solution, we next compared OS exhibited in spiking responses with that in subthreshold responses (i.e., residual membrane potentials after filtering out spikes). As shown by an example cell (Figure 1D), robust membrane depolarization responses were evoked by gratings at all testing orientations, although significant spiking responses were only observed for two orientations. Therefore, the orientation tuning of postsynaptic potential (PSP) response was much weaker compared to that of spiking response, although the two types of response exhibited the same optimal orientation (Figure 1E). In a total of 24 simple cells, similarly we found that spiking and PSP responses in the same cell exhibited essentially identical preferred orientations (Figure 1F). The orientation selectivity index (OSI, see Experimental Procedures) of spiking response was positively correlated with that of PSP response (Figure 1G).

05). We then acutely deleted FXR2 in NPCs in the DG of the adult WT mice using retrovirus signaling pathway that only infected dividing cells ( Liu et al., 2010 and Smrt et al., 2010) ( Figures S3E–S3H). Viral infection resulted in increased proliferation ( Figures S3I–S3M) and increased neuronal differentiation ( Figure S3N). Therefore, acute knockdown of FXR2 in adult NPCs results in phenotypes similar to those we observed in Fxr2 KO NPCs, both in vitro and in vivo. Taken together, our results provide further evidence that FXR2 plays a role in regulating the proliferation and differentiation of NPCs specifically in the adult

DG. To determine how FXR2 regulates NPCs in the DG, we first used real-time PCR-based neural stem cell pathway arrays to identify genes that exhibited altered expression levels in Fxr2 KO DG-NPCs relative to WT cells ( Figure S4A). Among the genes with >2-fold changes in Fxr2 KO DG-NPCs ( Figure S4B), we selected Shh (sonic hedgehog), Epacadostat clinical trial Notch2 (Notch gene homolog 2), Sox3 (SRY-box containing gene 3), and Noggin

for further analyses, due to their well-known functions in NPCs ( Lim et al., 2000, Ninkovic and Gotz, 2007, Palma et al., 2005, Solecki et al., 2001 and Wang et al., 2006). The up-regulation of Noggin in Fxr2 KO DG-NPCs was particularly interesting, because Noggin has been shown to promote the self-renewal of DG-NPCs, but not SVZ-NPCs ( Bonaguidi et al., 2008). FXR2 is known to bind mRNAs and regulate protein translation (Darnell et al., 2009 and Kirkpatrick et al., 2001). Using immunoprecipitation of FXR2 and its bound RNAs (RNA-IP), we confirmed that FXR2 bound to Noggin mRNA ( Figures 5A and 5B) but not to Shh, Notch2, or Sox3 mRNAs ( Figure S4C). In addition, biotin-labeled synthetic Noggin mRNA indeed bound FXR2 protein in NSC protein lysate, whereas an antisense control RNA did not ( Figure 5C).

Furthermore, on separately isolated DG-NPCs, we confirmed that Noggin mRNA levels were elevated in the Fxr2 KO DG-NPCs ( Figure 5D). The Rebamipide increased Noggin mRNA levels could be due to either increased gene transcription or increased mRNA stability. We treated WT and KO NPCs with actinomycin D to inhibit gene transcription and found that Noggin mRNA had a longer half-life in Fxr2 KO DG-NPCs than in WT cells ( Figure 5E; n = 3), while the half-life of Notch2, Shh, and Sox3 mRNA showed no significant difference ( Figures S4D–S4F). We then manipulated FXR2 levels in WT and Fxr2 KO DG-NPCs and found that acute knockdown of FXR2 in WT NPCs resulted in a longer half-life of Noggin mRNA, while exogenous FXR2 reduced the Noggin mRNA half-life in Fxr2 KO DG-NPCs ( Figure 5E). Therefore, FXR2 expression levels directly affect the stability of Noggin mRNA in DG-NPCs.