We took special care in choosing a reference montage for our analysis, as it has been shown that using a common reference can sometimes lead to specious Dasatinib manufacturer results when investigating properties of the phase (Schiff, 2005). The decision to use a bipolar montage was based on the assumptions that (1) the unwanted reference signal was recorded equally by each electrode in the bipolar pair and (2) each electrode in the pair measured complementary components of the same phenomenon (Zaveri et al., 2006). Assumption (1) is satisfied due to the physical setup of our recording device, and assumption (2) is valid due to the extremely localized measurements of the microwire electrodes.

If anything, we would be concerned that two adjacent microwires were placed so close together that they give exactly the same measurements and the bipolar pair would thus be useless. The relevant local information would be thrown away with the unwanted reference signal. We can see from the results presented here that this is not always the case, although it may account for the variability of results from each microwire bundle. In general, the use of a bipolar montage selleck products is the most conservative choice we can make; it guarantees that the signals used in our analysis are localized to a specific brain region, but it may also reduce the strength of the results due to the loss

of relevant behavioral information. Patients were presented with a 4 × 4 grid of face-down cards on a laptop computer screen and were told that there were eight pairs of matching cards (Figure 1A). When they used the mouse to click on a face-down card, the card flipped over and an image appeared. The goal was then to click on the matching card hidden among the other face-down images. After clicking on a pair of cards, matched pairs remained visible, while unmatched images flipped over again after approximately 1 s in order to be matched on a later turn. When all 16 cards were matched, a new puzzle was generated with randomly chosen images and locations. The game contained eight categories of images (e.g., faces, teddy bears, giraffes, watermelons, ice cream, shoes,

globes, and waterfalls), with six unique images in each category. Each subject completed two sets of ten puzzles. A set contained Dichloromethane dehalogenase 80 correct trials and 87.9 ± 20.1 incorrect trials, depending on how efficiently the patient completed the task. The experiment was run using the Psychophysics Toolbox in MATLAB. In our analysis, we draw a distinction between the mouse click on the first image of each pair (“first click”) and the next click on its potential match (“second click”). For the six subjects, the average time between the first and second click ranged from 1.2–2.1 s for the first set of ten puzzles, and it ranged from 1.0–1.7 s for the second set of puzzles. The average time between all clicks ranged from 1.7–2.3 s and 1.5–1.

In addition to Shh expression, we also found AF64α dose-dependent alterations in the expression of dopaminergic markers that resembled the distortions seen in Shh-nLZC/C/Dat-Cre mice ( Supplemental Results D and Figure S3E) indicating that altered signaling by cholinergic neurons contributes to the dopaminergic cell syndrome that we observe in the absence of Shh signaling from DA neurons ( Figure 2). These findings suggested that the transcription of Shh in DA neurons is activated in Shh-nLZC/C/Dat-Cre mice. Because the design of the Shh-nLZC/C allele leaves intact the promoter and most transcriptional enhancer regions of the native PD0332991 manufacturer Shh locus after Cre mediated

recombination we devised a qPCR based assay to test whether Shh expression was activated in DA neurons before and after ACh neuron degeneration in Shh-nLZC/C/Dat-Cre mice (for technical details see Supplemental Results A and Figure S1). Y-27632 solubility dmso We found an ∼5-fold and ∼4-fold increase in the transcription of the 5′-end of the Shh mRNA that can be expressed from the truncated Shh locus in Shh-nLZC/C/Dat-Cre at 4 weeks and 12 months of age, respectively. Thus, Shh-nLZC/C/Dat-Cre animals cease to produce signals that otherwise inhibit

Shh expression by mesencephalic DA neurons in the undisturbed brain ( Figure 7F). The identification of the receptor(s) on DA neurons why that transmit the signals that impinge on the regulation of Shh expression can inform on the nature

of the signals. We noted that the tissue specific ablation of the canonical receptor Ret, which can bind all members of the GDNF family of ligands, from DA neurons utilizing the same Dat-Cre allele also employed in the present study, resulted in alterations in dopaminergic marker gene expression, deficits in elicited DA release and late-onset, progressive DA neuron degeneration, (RetC/C/Dat-Cre mice) ( Kramer et al., 2007). We tested whether also the expression of Shh is altered in the vMB of RetC/C/Dat-Cre mice. We found an ∼6-fold upregulation of Shh expression in RetC/C/Dat-Cre mice compared to litter controls at 3 months of age prior to observable neurodegeneration ( Figure 7F). Taken together, our studies provide pharmacological and genetic evidence that ACh neurons of the striatum produce signals, which engage the canonical GDNF receptor Ret on DA neurons and repress the expression of Shh, and regulate the expression of multiple other genes in DA neurons. Our results reveal that mesencephalic DA neurons express Shh throughout life and demonstrate that DA neuron-produced Shh is necessary for the long-term structural and functional maintenance of mesencephalic DA neurons. Our studies, however, did not uncover any evidence for an autocrine mode of Shh signaling.

, 2009). Given that these adhesive contacts must be shed upon differentiation, we next investigated whether the BI-6727 pro-differentiation actions of Foxp4 might involve changes in N-cadherin expression or subcellular distribution. In transverse sections of the spinal cord, we noticed that there was a slight thinning of apical N-cadherin staining around the region of the pMN (Figure 3A, bracket). This difference was more clearly revealed by imaging the apical surface of the neuroepithelium in an open book preparation, which showed distinct bands of N-cadherin staining corresponding to the different

progenitor domains along the dorsoventral axis (Figures 3B–3E and S5A–S5J). N-cadherin was strikingly reduced wherever Foxp4 was present (Figures 3B and 3D–3F; averaged correlation R2 = −0.722). This antithetical pattern was specific to Foxp4 and N-cadherin as there was no correlation between the expression of Foxp4 and other AJ

components such as aPKCζ or the NPC marker Sox2 ( Figures 3B, 3C, and S5B–S5L). Under conditions of Foxp4 misexpression, the electroporated spinal cords displayed a dramatic loss of N-cadherin protein and disruption in the ultrastructure of the neuroepithelium (Figures 3G, 3K, 3M, and 3Q). These changes coincided with an aberrant distribution or loss of other AJ components learn more including β-catenin, f-actin, aPKCζ, and Par3 (Figures 3H, 3I, 3N, and 3O, and data not shown) and cytoplasmic unless accumulation of Numb (Figures 3L, 3M, and 3R). The radial morphology of NPCs was also severely disrupted (Figures 3H, 3I, 3N, and 3O), and markers of dividing cells such as BrdU incorporation and phosphohistone H3 staining were reduced (data not shown). Nonetheless, integrin-laminin interactions at the basolateral membrane remained intact (Figures 3J and 3P), suggesting

that the effects of Foxp4 misexpression are primarily directed to apical attachments. Identical results were seen with misexpression of Foxp2 and Foxp1 (Figure S3), indicating that all of the Foxp proteins have the capacity to repress N-cadherin expression and disrupt AJs under these conditions. The combined knockdown of Foxp2 and Foxp4, in contrast, led to an ∼1.5–2-fold upregulation of N-cadherin mRNA and protein within the pMN and extensive accumulation of Numb at the apical membrane of these cells (Figures 3S, 3T, and S5M–S5Q). The effects of the shRNA constructs were specific, as the knockdown phenotype was completely reversed by coelectroporation of a Foxp4 expression vector, often resulting in the Foxp4 misexpression phenotype (Figures S2J–S2R). Together, these data indicate that Foxp2 and Foxp4 play a crucial role suppressing the expression of N-cadherin and disassembling neuroepithelial AJs (Figure 3U).

These properties, which we will address below, include (1) cross-orientation

suppression, (2) contrast invariance of orientation tuning width, (3) contrast-dependent changes in response timing and in temporal frequency preference, and (4) the mismatch between measured orientation tuning and the tuning predicted by a simple cell’s receptive field organization. Uncovering the origin of these properties has proven to be one of the keys to selleck compound understanding the nature of the cortical computation. One comprehensive solution to the origin of simple cell nonlinearities was suggested by psychophysics: in the tilt aftereffect illusion, the perceived orientation of a vertical stimulus is shifted away from vertical after prolonged viewing of a slightly oblique stimulus. This result was interpreted to mean that intracortical inhibition, specifically inhibition between cortical neurons of different preferred orientations, sharpened orientation tuning

or even created it de novo (Blakemore and Tobin, 1972). This proposal was strengthened by pharmacological experiments: cortical application of GABAA antagonists cause a broadening of orientation tuning (Sillito, 1975). Cross-orientation inhibition, a form of lateral inhibition (Hartline, 1949), but in the orientation domain rather than the spatial domain, is considered a natural extension of similar mechanisms either observed or proposed to operate throughout the brain. Because of the columnar organization of orientation preference in the cortex, the orientation domain translates into the spatial domain on the cortical surface. http://www.selleckchem.com/products/ON-01910.html Cross-orientation inhibition can then emerge from simple, spatially defined rules of cortical connectivity. Cross-orientation inhibition has been proposed to operate in several distinct modes, depending on the orientation dependence and amplitude of inhibitory interconnections.

In attractor models, those feedback inhibition forms a set of multistable attractors (Ben-Yishai et al., 1995 and Somers et al., 1995), in which the width of orientation tuning of cortical cells is determined by the lateral extent of cortico-cortical connections. In recurrent models, recurrent excitatory connections amplify feedforward inputs in a way that is sculpted by lateral inhibitory connections (Douglas et al., 1995). Here again, the width of tuning and other aspects of cortical responses are set by intracortical rather than thalamocortical interconnections. In balanced models, strong recurrent excitation and inhibition are thought to balance one another tightly (van Vreeswijk and Sompolinsky, 1998). In addition to explaining many aspects of simple cell behavior, this balance can explain the large variability of cortical spiking responses (Shadlen and Newsome, 1998). In push-pull models, cross-orientation inhibition arises from feedforward inhibition from simple cell-like inhibitory interneurons (Troyer et al., 1998 and Troyer et al.

These results indicate a neuronal site-of-function of cysl-1 in regulating the egl-9/hif-1 pathway to modulate the O2-ON response. We used BLASP to search the NCBI protein database and found many CYSL-1 homologs belonging to the cystathionine-beta synthase/cysteine PD0325901 cell line synthase (CBS/CS) family of the fold type-II pyridoxal-5′-phosphate (PLP)-dependent proteins in diverse species ranging from bacteria to humans (Figures 5A and S5A). The cysl-1(n5515) allele we isolated from the rhy-1(n5500) suppressor screen converted glycine 183 to arginine ( Figure 5A, Table 1B). Strikingly, this glycine is 100% conserved among the cysl-1 homologs of all species examined (bacteria,

yeast, flies, zebrafish, mice, and humans) and is positioned at the core of a motif sequence crucial for binding to the obligate cofactor

PLP ( Aitken et al., 2011) ( Figures 5A and S6C). Interestingly, one of the CYSL-1 paralogs is the HIF-1 target gene K10H10.2, click here indicating a possible feedback regulation of this gene family. We raised a polyclonal CYSL-1 antibody and found reduced levels of steady-state CYSL-1(n5515) proteins in soluble fractions of C. elegans and bacterial homogenates compared to those of wild-type CYSL-1 ( Figures 5B and S5B). The introduction at residue 183 of arginine, which has a long protruding hydrophilic side chain ( Figure S6E), could disrupt binding to PLP and render the protein improperly folded and unstable. n5521, n5522, and n5537 mutants similarly showed

reduced levels of CYSL-1 ( Figures 5B and S5B, S6C–S6F). We studied recombinant CYSL-1 proteins purified Electron transport chain from E. coli and found that CYSL-1 exhibited properties typical of type-II PLP-dependent proteins ( Figures S5D–S5G). We tested several biochemical reactions that had previously been associated with other PLP-dependent CBS enzymes and cysteine synthases ( Aitken et al., 2011 and Mozzarelli et al., 2011). While assays for O-phosphoserine sulfhydrylase, cyanoalanine synthase, and cystathionine beta-synthase failed to yield significant enzymatic activities, CYSL-1 exhibited activity as an O-acetylserine sulfhydrylase (OASS), converting OAS and sulfide into L-cysteine and acetate ( Figures 5C and 5D). However, the Michaelis constant KM for sulfide (4.2 mM) of purified CYSL-1 was at least an order of magnitude higher than those of bona fide cysteine synthases, CYSL-1 homologs from bacteria and plants ( Figure 5E), suggesting that the cysteine synthase activity of CYSL-1 might be insignificant physiologically in vivo and dispensable for regulating the egl-9/hif-1 pathway. cysl-1(n5519) mutations suppressed HIF-1 target expression and restored the O2-ON response of rhy-1(n5500) mutants, yet the CYSL-1(n5519) mutant protein, with the abnormal lysine (R259K) residue on its surface far from the active site ( Figure S6F) exhibited levels of OAS sulfhydrylase activity similar to that of wild-type CYSL-1 ( Figures S6A and S6B, Table 1B).

After being habituated with two objects for two consecutive days in the arena, Fmr1 KO mice failed to recognize the novel object introduced by replacing one of the familiar objects on the third day of the test.

In contrast, WT and dKO mice were able to distinguish the new object with high fidelity when tested under same conditions. Interestingly, S6K1 KO mice were impaired in their ability to discriminate between familiar and novel objects. These findings indicate that removal of S6K1 in Fmr1 KO mice can restore appropriate novel object recognition memory. We also examined whether the four genotypes showed differences in their ability to learn and recall a task and UMI-77 selleck compound their flexibility in modifying their responses when the task was changed using a water-based Y-maze test. All four genotypes responded to training and learned the test with comparable efficiency on the first day (Figure 6D). Memory

recall for the arm location of the hidden platform also was robust on the day after training for all four genotypes, indicating that there were no deficits in long-term memory (Figure 6D). When the platform location was reversed, Fmr1 KO mice displayed impairments in reversal learning, requiring an additional 15 trials to meet criterion as compared to WT, S6K1 KO, and dKO mice ( Figures 6D and 6E). These findings suggest that upregulation of S6K1 signaling plays a key role in behavioral inflexibility in FXS model mice. Fmr1 KO mice also have been reported to show increased ambulatory behavior in the open field test ( Spencer et al., 2005), which we reproduced in our behavioral cohort ( Figures S6A and S6B). In contrast, S6K1 KO littermates exhibited significantly decreased exploration and preferred the peripheral

areas versus the center of the arena. Interestingly, dKO mice displayed open field exploration indistinguishable all from the Fmr1 KO mice, indicating that the deletion of S6K1 did not correct increased ambulatory behavior in FXS model mice. In addition, marble-burying behavior, a phenotype that has been used to model obsessive-compulsive behavior in mice, was enhanced in Fmr1 KO mice compared to WT littermates. We found that Fmr1 KO, S6K1 KO, and dKO mice buried a higher number of marbles compared to WT littermates ( Figure S6C). These findings indicate that the enhanced repetitive behavior of Fmr1 KO mice is not rescued by the abrogation of S6K1. FXS patients display neuroendocrine dysfunction that is reflected in a generalized increase in total body weight, macrocephaly, and enhanced stature (Penagarikano et al., 2007). Another frequently associated feature is macro-orchidism (enlarged testicles), first observed in male patients immediately after attaining puberty (Hagerman et al., 1983).

A growing number of investigators who were trained as basic neuroscientists have become engaged in projects that relate to specific brain diseases and disorders. In part, this reflects pressure from NIH and other funding agencies to promote research related to the core mission of improving health. However, I believe that first Selleckchem Vorinostat and foremost this trend reflects the tremendous progress in the field that enables meaningful attacks on key disease-related problems,

using a variety of animal models as well as human patient populations. In my case, I was drawn into disease-related research about a decade ago when a serendipitous opportunity arose to collaborate in applying our cortical cartography tools (Caret software) to a study of children with Williams syndrome. We identified dozens of cortical folding abnormalities in Williams syndrome and hypothesized that these folding abnormalities might be caused by underlying circuit abnormalities GABA receptor function via the aforementioned tension-based folding hypothesis (Van Essen et al., 2006). Soon thereafter, I struck up a collaboration with pediatric neurologists Terrie Inder and Jeff Neil to study cortical development using structural MRI scans in infants, including those born prematurely. This is extremely important clinically, because of the disturbingly high incidence of various behavioral disorders in children

born prematurely (Bos and Roze, 2011). It is also provides a fascinating window on a period of rapid development, when gyrification is in full swing and the cortex is expanding rapidly. In comparing healthy term-born infants to adults, we discovered that postnatal cortical expansion is strikingly nonuniform, with the greatest expansion occurring in lateral temporal, prefrontal, and parietal regions that are implicated in cognitive function (Hill et al., 2010). Earlier sections have already emphasized the importance of studying nonhuman primates, especially the macaque, as model systems for better understanding the human brain. However, human cerebral

cortex is not only larger (by 10-fold) and far more convoluted than the macaque, but it is far from being a scale model even after smoothing out the wrinkles (see Figures 2B and Cediranib (AZD2171) 2C). The need for objective and quantitative comparisons in the face of large interspecies differences poses an interesting cartographer’s challenge. Years ago I realized that the surface-based registration methods that we had developed for within-species registration could be adapted to registration between species. By assigning landmarks to areas known as suspected to be homologous in the macaque and human, registration constrained by these landmarks indicates that lateral temporal, parietal regions expanded 20-fold or more in the human lineage compared to the macaque, whereas early sensory areas expanded far less (Van Essen and Dierker, 2007; see also Chaplin et al., 2013).

Interestingly,

p150WT-HA expressed in motor neurons is dramatically enriched within NMJ TBs ( Figure 2G, arrows), in addition to its expected localization along axons ( Figure 2G, asterisks) and in the cytoplasm. We observe that the TB localization of p150WT-HA is apparently greatest within the center of the TB, just distal to where expression of the microtubule-associated check details protein Futsch becomes undetectable ( Figure 2G). p150WT-HA is also enriched at sites of microtubule loops, which are thought to be enriched in microtubule plus ends ( Figure 2G, arrowheads; enlarged in Figure S3C) ( Roos et al., 2000). To determine whether microtubule plus ends are also enriched at TBs, we expressed a microtubule plus-end marker, the kinesin motor domain fused to GFP (KhcHead:GFP) ( Clark et al., 1994), in motor neurons. Interestingly, we see at the NMJ that KhcHead:GFP is predominantly

localized to the TB ( Figures 2H and S3D) and, similar to p150WT-HA localization, is enriched within the middle of the TB ( Figure 2H, inset). We also observe a similar enrichment of the microtubule plus-end marker EB1:GFP at this location ( Movie S3). These data suggest that wild-type p150Glued is enriched at microtubule plus ends of terminal boutons. Because p150WT-HA is localized within NMJ TBs, we next investigated the morphology of the presynaptic nerve terminal in Glued mutants. Anti-HRP labels the presynaptic membrane at the Drosophila NMJ and binds to neuron-specific transmembrane glycoproteins such as FasII ( Desai et al., 1994). Interestingly, we observe Vemurafenib datasheet intense anti-HRP staining within TBs of GlG38S and GlG38S/GlΔ22 NMJs ( Figure 3A), suggesting that neuronal membranes accumulate at these presynaptic termini. Similar to the TB swelling we observed in GlG38S/GlΔ22 mutants, the anti-HRP phenotype is more severe in distal abdominal segments than in proximal segments ( Figures 3A and

3D). Approximately 75% of NMJs from distal segments of GlG38S and GlG38S/GlΔ22 larvae display accumulation of anti-HRP staining within TBs, whereas 17-DMAG (Alvespimycin) HCl only ∼15% of control NMJs have any accumulation of anti-HRP staining within TBs ( Figure 3D). Similarly, overexpression of p150G38S in motor neurons (using D42-GAL4) causes a dramatic accumulation of anti-HRP immunoreactivity in large puncta specifically located within the TB, demonstrating that p150G38S can act in a dominant-negative fashion when overexpressed in neurons ( Figures 3B–3D). Because anti-HRP labels presynaptic transmembrane proteins, these data suggest that membrane-bound vesicles accumulate within TBs of GlG38S NMJs. We next crossed D42-GAL4, UAS-p150G38S (D42 > p150G38S) flies to flies that express fluorescently tagged markers that label distinct membrane-bound compartments under control of the UAS promoter. Colocalization of the membrane marker mCD8:GFP with anti-HRP in terminal boutons of larvae expressing p150G38S suggests that these anti-HRP positive structures are membrane bound ( Figure 3C).

To increase the urban and rural sub-region rates to 2011 estimates, we select a random set of households to also vaccinate. In the intervention scenarios, to scale up the coverage rates, the model makes additional households vaccination compliant. The method of selecting these extra households varies across scenarios (e.g., random or targeted by state and region). The model was programmed in C++. Analysis variables fall into four categories, which consider the intervention’s associated effect on disease burden, intervention costs, cost-effectiveness, and financial impact. The effect on disease burden

includes both deaths and disability-adjusted life years (DALYs) averted (we discount at 3% and use uniform age-weights that value any extra year of life equally). Cost-effectiveness is measured by dollars per DALY averted incremental to the baseline scenario. The financial impact measures follow Verguet et al. [23] and include the Obeticholic Acid datasheet out-of-pocket (OOP) expenditure averted from the baseline scenario, which measures the savings of the population that result from the intervention, and the money-metric value of insurance, which measures the value of protection from expenditure on disease treatment

(including the costs of seeking care). The money-metric value of insurance here differs slightly from Verguet et al.’s analysis. Our analysis period is one year as we study a cross-section of the under-five population, while they study a birth cohort, which is susceptible to disease over the first five years of life. Given this, we include only one year of disposable income in the calculation INK 128 order as opposed to five years. Additionally, we evaluate the value of insurance of an intervention with respect to the baseline by subtracting one from the other. Resminostat We analyze health and financial burden alleviated across India by wealth quintile, state, and rural versus urban areas. To quantify the uncertainty of the model, we conduct a 100-simulation Latin hypercube sampling (LHS) sensitivity analysis over a plausible range of the input parameters (Table 1). For each

disease, the parameters analyzed include the incidence, CFR, vaccine efficacy, vaccine cost, and treatment cost. Ninety-five percent uncertainty ranges for our mean estimated outcomes are calculated on the basis of this sensitivity analysis and reported in parentheses. In the baseline, immunization coverage is 77% for DPT3, 82% for measles, and there is no coverage for rotavirus. From DLHS-3 data, we find that baseline coverage increases by wealth for DPT3 and measles. The rural-to-urban immunization coverage ratio is 1.09 for DPT3 and 1.05 for measles (Fig. 1, row 1). Baseline DPT3 coverage is lowest in Arunachal Pradesh and Uttar Pradesh where 53% and 55% of under-fives are vaccinated (Fig. 2, column 1). Another nine states vaccinate less than 80% of their children; all of them are relatively poor states, with the exception of Gujarat (77% coverage). Eight states have DPT3 coverage above 90%.

As depicted in Fig. 3A, a clear upregulated pattern of expression of CD40, CD80 and CD86, but not CD40L, can be seen on the surface of CD11c+PDCA-1+

cells obtained from the LN. In contrast, we detect only the upregulation of CD40 on CD11c+PDCA-1+ splenocytes at day 10 after infection (Fig. 3B). In addition, we also stained LN and spleen cells for CD11c expression in conjuction with CD8α in addition to the activation markers CD40, CD40L, and CD86 at different times after infection. A limited pattern of upregulation of expression of find more CD86 can be seen on the surface of CD11c+CD8α+ cells collected from the LN or spleen on days 3–7 following infection (Fig. 4A and B). Similar analyses were also conducted for CD11C+CD8a− cells collected

from the spleen and LN, but we did not detect an upregulation of expression of the activation markers CD40, CD40L, CD80, or CD86 at any time point from 3 to 30 days in the spleen or LN (data not shown). To determine whether indeed CD11c+PDCA-1+ cells could present antigen for specific CD8 lymphocytes, we purified CD11c+PDCA-1+. After sorting the cells from naïve or 5-day infected Lonafarnib research buy LN cells, we obtained cells that were 95.3 and 83% pure as determined by the PDCA-1 marker (Fig. 5A and B, respectively). For some unknown reason, during the purification process, some cells become negative for the marker for CD11c marker but still

retained the PDCA-1 marker. The PDCA-1+ cells obtained from mice that were infected expressed significantly higher amounts of MHC-II-IAb and CD80 (Fig. 5C and D, respectively). PDCA-1+ secondly cells were used to stimulate purified CD8+ splenic cells obtained from T. cruzi infected mice. As shown in Fig. 5E, IFN-γ producing cells were detected only when CD8+ were incubated with PDCA-1+ cells obtained from infected mice. The fact that CD11c+ cells from the spleen exhibit a limited activation phenotype suggested that perhaps most of the specific T cells found in the spleen might not be primed there. If this assumption is correct, the re-circulation of T cells could account for the CD8+ T-cell mediated functions detected in this organ. To test whether lymphocyte re-circulation was responsible for the immune response observed in the spleen, we treated infected mice with FTY720. This immunosupressive drug inhibits S1P1 signalling, thus efficiently blocking re-circulation of naïve and activated T cells from the LNs into peripheral tissues, thereby preventing development peripheral T-cell responses [27], [28] and [29]. Mice were infected with T. cruzi parasites and FTY720 or diluent were administered on the same day of challenge and every 2 days thereafter as described in Section 2.