Instead, one may ask how large the cortical region is that generates the LFP. Several recent experimental studies have addressed this question (Kreiman et al., 2006, Liu and Newsome, 2006, Berens et al., 2008a, Katzner et al., 2009 and Xing et al., Abiraterone research buy 2009) but have reported different results ranging from a few hundred micrometers (Katzner et al., 2009 and Xing

et al., 2009) to several millimeters (Kreiman et al., 2006). How can the results be so different? One possibility is that the LFP reported in various experiments stems from different types of neuronal populations or that the electrodes have been placed differently. Moreover, different stimulation paradigms have been used, likely resulting in different levels of correlations between the synaptic currents providing the recorded LFP. It has long been suggested that the LFP is dominated by synchronously driven dendritic

input on pyramidal cells (Mitzdorf, 1985), but it has until now been unclear how the amount and spatial extent of correlations in synaptic activity influence the LFP. In the present study, we investigate various key factors determining the size of the region an LFP electrode can “see,” in particular, the neuronal morphology, synaptic distribution, level of correlation in synaptic activity, and the position of the recording electrode. We use a biophysical forward-modeling Panobinostat cell line approach to address these questions (Holt and Koch, 1999, Pettersen et al., 2008, Pettersen and Einevoll, 2008 and Lindén et al., 2010) and simulate the LFP signal from synaptically activated populations of morphologically reconstructed cortical cells. The LFP amplitude generally increases with increasing radius of the model population, but typically it flattens out beyond a certain radius, here termed the spatial reach. For uncorrelated synaptic activity, we find this spatial reach to be only a few hundred micrometers, implying that the recorded LFP is generated by a small population of neurons surrounding the electrode. This result is in line with findings in recent experimental studies ( Katzner

et al., isothipendyl 2009 and Xing et al., 2009). However, for particular synaptic distributions onto pyramidal cells, we find the reach of the LFP to be much larger and depend strongly on the level and spatial scale of correlations in the synaptic input, putatively explaining the disparate results reported in other experimental studies ( Kreiman et al., 2006, Liu and Newsome, 2006, Berens et al., 2008a, Katzner et al., 2009 and Xing et al., 2009). Our simulation findings are supported by analytical results using a simplified, yet as it turned out, accurate model of LFP generation. This model encapsulates the dependence of the population LFP on the spatial decay of single-neuron LFP contributions and correlation of synaptic input.

We next examined whether the chronic opiate-induced morphological change was correlated with changes in DA neuronal excitability. Proteases inhibitor We found that chronic morphine-treated mice, compared with sham-treated mice, exhibited an increase in the spontaneous firing rate of VTA DA neurons in brain slices (Figure 1C). This effect was not dependent on residual morphine in the slice, since blockade of opioid receptors with naloxone did not affect cell excitability

(Figure 1C). Moreover, the inclusion of a low dose of morphine (5 μM) in the bath solution to prevent “withdrawal” in the slice did not alter DA neuron firing rate (Figure 1C). Given the observations that chronic morphine decreases the size of VTA this website DA neurons, but concomitantly increases their excitability, it was important to determine whether net DA output from VTA is altered. We examined levels of extracellular DA in nucleus accumbens (NAc) in vivo, widely considered a key determinant of reward (Hyman et al., 2006). In opposition

to the increased firing rate, we found that chronic morphine dramatically decreased electrically evoked DA output in NAc of rats as measured by fast-scan cyclic voltammetry (Figure 1D). This reduction in DA output from VTA DA neurons supports the notion that the reduced soma size of the neurons, induced by chronic morphine, correlates with functional output, consistent with the reward next tolerance induced by chronic morphine under these conditions (Russo et al., 2007). Next, we examined a possible relationship between the increase in VTA DA neuron firing rate and soma size decrease, with the hypothesis that the increased firing rate per se induces changes in soma size. We virally overexpressed

a dominant-negative K+ channel subunit (dnK, KCNAB2-S188A, R189L) locally within VTA; we showed previously that this mutant channel increases the firing rate of VTA DA neurons (Krishnan et al., 2007). Overexpression of dnK was sufficient to decrease the surface area of VTA DA neurons (Figure 2A). To obtain the converse type of information, we virally overexpressed wild-type Kir2.1 in VTA, which we showed decreases DA neuron firing rate (Krishnan et al., 2007). While overexpression of Kir2.1 alone did not affect VTA DA soma size (data not shown), it completely blocked the ability of chronic morphine both to decrease soma size (Figure 2B) and to increase DA neuron firing rate (Figure 2C). These findings support our hypothesis that the morphine-induced increase in VTA DA neuron excitability is both necessary and sufficient for mediating the decrease in soma size. Given the increase in VTA DA neuronal firing rate observed in response to chronic morphine, we examined possible underlying mechanisms. One possibility is that morphine, by downregulating AKT activity in these neurons (Russo et al.

Large cell clusters were seldom observed, suggesting that these interactions are weak or limited by unknown factors. Within the CNS, Ptp10D is expressed only on axons (Sun et al., 2000; Tian et al., 1991; Yang et al., 1991) (Figure 4A). It is also expressed on the apical surfaces of tracheal cells (Jeon and Zinn, 2009) (Figure 4C). Antibody against Sas has been widely used as a selective marker for apical cell surfaces (Schonbaum et al., 1992; Wodarz et al., 1995). Sas is probably expressed at some level on the apical surfaces of all cells of epithelial origin, including Carfilzomib cost neurons and glia. Most or all cell bodies and axons in the VNC appear to stain with anti-Sas. Sas is expressed at higher levels on some axons than

on others, in particular on the intersegmental and segmental nerve roots and on two longitudinal bundles on each side of the CNS (Figure 4B). Figure S4 demonstrates that ensheathing cells on the dorsal surface of the CNS express Sas. These are the perineurial, subperineurial, selleck kinase inhibitor interface, and channel glia (Ito et al., 1995). In the tracheal

system, the patterns of staining with anti-Ptp10D and anti-Sas are essentially identical (Figures 4C and 4D) (Jeon and Zinn, 2009). Although lethal ethyl methane sulfonate (EMS) mutations in sas were isolated many years ago, little is known about the functions of the sas gene. sas mutants do not exhibit embryonic lethality, but their postembryonic growth is stunted and they usually die as second instar larvae (hence the name). Third instar escapers have convoluted tracheae ( Schonbaum et al., 1992), but we were unable to find any tracheal defects in embryos. To evaluate whether sas mutants have CNS phenotypes,

we examined embryos homozygous for a strong EMS mutation, sas15, or transheterozygotes bearing this mutation over Df(3R)ED5221, which removes the entire sas gene. We sequenced the sas gene on the sas15 chromosome, and found that sas15 introduces a stop codon after aa 642, which is N-terminal to all conserved Sas domains. These data indicate that sas15 is likely to be a null mutation. To visualize CNS axons, we used mAbs BP102 and Phosphoprotein phosphatase 1D4. BP102 labels most or all CNS axons, producing a ladder-like pattern, with two commissural tracts in each segment and two longitudinal tracts extending the length of the embryo (Seeger et al., 1993) (Figures 5B, 5D, and 5F). 1D4 recognizes the transmembrane form of FasII (Vactor et al., 1993), and stains pioneer axons at stage 14 (Figure 5A). By late stage 16, 1D4 staining within the CNS is restricted to three distinct longitudinal axon bundles on each side of the VNC, and no staining is seen on commissural axon bundles that cross the CNS midline (Figure 5E). sas15/Df transheterozygotes appear normal at stage 14 ( Figures 5G and 5H), but display weak longitudinal axon defects at stage 16. The outer 1D4 axon bundle is interrupted and the other bundles appear slightly irregular ( Figures 5I and 5K).

During development of the sensory epithelium in the cochlea, the Cdki, p27kip1 is an early marker of the part of the presumptive sensory region that will generate the hair cells and support cells ( Chen and Segil, 1999) and deletion of p27kip1 leads to an extension in the normal developmental limit in proliferation of cells in the cochlea ( Kil et al., 2011, Lee et al., 2006 and Löwenheim et al., 1999). Although these experiments demonstrated that p27kip1 is an important developmental regulator of support

cell proliferation, recently it was shown that deletion of p27kip1 in adult animals also causes support cells to enter the mitotic cell cycle ( Oesterle et al., 2011), albeit in small numbers, indicating Sunitinib purchase that p27kip1 Caspase activation is one of the factors required in mature

mice to maintain mitotic quiescence in the support cells. Taken together, the results suggest that methods to stimulate proliferation in the mammalian inner ear epithelia might be possible through manipulation of a combination of known pathways. However, even though some support cells proliferate in the postnatal cochlea in the p27kip1 knockout mice, very few, if any, generate mature new hair cells as they would during regeneration in nonmammalian vertebrates; rather, the proliferating cells appear to generate additional support cells, or else they undergo apoptosis. This leads to the second main difference between the nonmammalian vertebrates and mammals: the support cells of the auditory sensory epithelium of nonmammalian vertebrates have the capacity to transdifferentiate into hair cells, while mammalian cochlear support cells do not. What factors enable the support cells of nonmammalian vertebrates to differentiate into hair cells after damage? Studies of the factors that control the fates of hair cells and support cells during regeneration have focused on the developmental regulators of hair cell determination/differentiation: the bHLH transcription

factor, Atoh1, and the Notch pathway (Cafaro et al., 2007, Daudet et al., 2009 and Stone and Rubel, 1999). Atoh1 is a critical transcription before factor for the specification of the hair cells during development (Figure 3), while Notch signaling has both a “prosensory” role and acts in a more conventional lateral inhibitory manner to regulate the ratios of hair and support cells. (Brooker et al., 2006, Kiernan et al., 2001, Kiernan et al., 2006, Adam et al., 1998, Brooker et al., 2006, Haddon et al., 1998, Kiernan et al., 2005 and Zine and de Ribaupierre, 2002). In the normal adult vestibular organs in birds, Atoh1 is expressed in scattered cells throughout the epithelium, suggesting a continued requirement for specification during the ongoing hair cell production in these organs (Cafaro et al., 2007). By contrast, in the normal avian adult BP, there is no Atoh1 expression; however, after hair cell damage a number of cells express Atoh1 and Notch pathway genes (Cafaro et al.

Again, this comparison revealed no significant difference (p = 0.14, t = 1.55). Taken together, these analyses suggest that the representation of sensory evidence as well as unspecific BOLD responses in early sensory areas did not change significantly over the course of learning. So far we have shown that (1) the predictions IWR-1 cost of an adapted reinforcement learning model correlate with learning-related changes in orientation discrimination performance over time and (2) that the model-derived DV, which builds the basis for perceptual

decisions, is coded in the medial frontal cortex. However, because alternative learning models would also predict similar increases in DV over learning, in the following analyses we provide further evidence for the proposed reinforcement learning mechanism. Evidence for Rescorla-Wagner-like updating in the reward-learning literature originally came from Z-VAD-FMK in vitro the observation of signed reward prediction error signals in dopamine neurons ( Bayer and

Glimcher, 2005 and Schultz et al., 1997). In human fMRI studies, however, prediction error signals have been identified in the ventral striatum, a target area of dopaminergic midbrain neurons ( Kahnt et al., 2009, McClure et al., 2003, O’Doherty et al., 2003 and Pessiglione et al., 2006). Thus, to provide further evidence for a reinforcement learning process in the current perceptual learning task, we regressed the signed prediction errors from the model against the feedback-locked BOLD signal in each voxel (see Experimental Procedures). We identified significant (p < 0.0001, k = 5) correlations between model-derived prediction errors and activity in the left ventral striatum ([-9, 0, −3],

t = 4.77; ADP ribosylation factor Figure 7A), the bilateral anterior insular cortex extending into the lateral OFC (left BA 47 [-33, 21, −3], t = 5.56; right BA 47 [30, 21, −6], t = 6.49), the dorsolateral PFC (right BA 9 [54, 15, 36], t = 5.17), as well as the dorsomedial prefrontal cortex including the ACC (BA 32 [0, 27, 42], t = 5.81; Figure 7B; see Table S3 for complete results). This shows that the key learning variable of our computational model, namely the signed reward prediction error, is coded in the activity of reward-related regions such as the ventral striatum, providing further evidence for a reinforcement learning process in perceptual learning. In a second step, we aimed to confirm that the learning-related changes in DV are indeed related to an updating mechanism that is based on signed prediction errors as proposed by our model. Thus, the same region in the ACC where activity patterns track perceptual learning-related changes in DV should also process reward prediction error signals.

The changes in taxonomy have, however, also contributed to changing the appearances in the inventory. Most of the species recorded as Candida in the former list have been transferred to other genera or included under the teleomorphic name ( Table 3). Recently, it has been suggested by many mycologists that only one name should be given to any fungus, as is already done in Zygomycota. Thus it would be preferred to refer to the most well-known species as Saccharomyces cerevisiae (the teleomorphic and holomorphic name), rather than the anamorphic name Candida robusta. According to present rules as guided by the International Code of Botanical Nomenclature

5-FU concentration Article 59, fungi in Ascomycota and Basidiomycota can have two names; one for the teleomorph and holomorph, which is recommended, and one for the anamorphic state. Candida

famata is the anamorph of Debaryomyces hansenii. Candida utilis, used for single mTOR inhibitor cell protein production, should be called Cyberlindnera jadinii. Williopsis mrakii (= Hansenula mrakii) is now also included in the genus Cyberlindnera as C. mrakii. Saccharomyces unisporus has been transferred to Kazachstania unispora, and Candida holmii has also been transferred to Kazachstania as K. exigua. Candida krusei is now called Pichia kudriavzevii. Candida kefyr (= Candida pseudotropicalis) is placed in Kluyveromyces marxianus. Candida valida is now called Pichia membranefaciens and finally Saccharomyces florentinus is now called Zygotorulaspora florentina ( Table 3; Boekhout and Robert, 2003 and Kurtzman et al., 2011). Regarding Candida, many additional species have been suggested for beneficial use in foods, including C. etchellsii, C. intermedia, C. maltosa, C. versatilis and C. zeylanoides. Teleomorphic states are not known for these species. Other species recently suggested include Clavispora lusitanae, Cystofilobasidium

infirmominiatum, Dekkera bruxellensis, Hanseniaspora uvarum, Kazachstania turicensis, Metschnikowia pulcherrima, Pichia occidentalis, Rhodosporidium sp., Saccharomyces pastorianus, Saccharomycopsis fibuligera, Olopatadine Saturnisporus saitoi, Sporobolomyces roseus, Torulaspora delbrueckii, Trichosporon cutaneum, Wickerhamomyces anomalus, Yarrowia lipolytica, Zygosaccharomyces bailii, and Z. rouxii. In the current update of the inventory of microorganisms, we tend to be conservative and only include species with a well-documented technological benefit. One example is Dekkera bruxellensis (anamorph Brettanomyces bruxellensis), which was formerly regarded as a spoiler of beer (and wine). However, it is used for production of Belgian Lambic-Geuze beer. D.

, 2006). Zinc has also been reported to inhibit native and recombinant KARs. selleck kinase inhibitor Zinc inhibition of KARs is subunit dependent, with KARs containing GluK4 or GluK5 subunits being more sensitive, IC50 ∼1–2 μM, than GluK1-GluK2, IC50 ∼70 μM (Mott et al., 2008). Despite the proposed presynaptic function of GluK3-containing KARs at hippocampal mossy fiber synapses, which are highly enriched in vesicular zinc, modulation of these receptors by zinc has not yet been

addressed. In this study, we show that zinc at micromolar concentrations potentiates recombinant GluK3 receptor currents evoked by glutamate. Zinc markedly slows receptor desensitization and increases apparent affinity for glutamate. By analysis of chimeric GluK2/GluK3 KARs and of GluK3 bearing selected point mutations, we mapped the zinc binding domain to the S2 segment of the LBD, in a region forming Erastin the interface between two GluK3 subunits in an LBD dimer assembly. Crystallographic studies for GluK3 LBD complexes with both glutamate

and kainate revealed that zinc ions bind at multiple sites formed by aspartate, histidine, and glutamate residues, which are present in both the upper and lower lobes of the LBD. Based on these crystal structures, a GluK3 LBD dimer model was generated by superposition of GluK3 monomers on previously solved KAR LBD dimers. This identified D730 as the dimer partner component of the binding site underlying zinc potentiation, together with D759 and H762 from the adjacent subunit. Based on these structure-function Adenosine studies and on modeling of

KAR activity, we show that zinc plays a very distinct role in GluK3-KARs by stabilizing the LBD dimer assembly, thereby reducing desensitization. Given the proposed presynaptic localization of GluK3 close to zinc-containing synaptic vesicles, zinc may be an endogenous allosteric modulator for native GluK3-KARs. We tested the effect of zinc on currents activated by fast application of glutamate on lifted HEK293 cells transfected with GluK3 cDNA. Currents evoked by sustained applications (100 ms) of 10 mM glutamate, a concentration close to the EC50 value for GluK3 (Perrais et al., 2009a; Schiffer et al., 1997), were markedly enhanced with preapplication of 100 μM zinc (Figure 1A; 193% ± 38% of control amplitude, n = 17), and this potentiation was rapidly reversible upon removal of zinc. In contrast to GluK3 potentiation, and as previously reported in Xenopus oocytes ( Mott et al., 2008), zinc reversibly inhibited GluK2 currents at all concentrations tested ( Figures 1A and 1D), with an IC50 of 102 ± 11 μM and a Hill coefficient (nH) of 1.1 ± 0.1 (n = 4–9). Because a glutamate concentration of 10 mM is saturating for GluK2 ( Perrais et al., 2009a), this could mask a potentiating effect of zinc. However, currents evoked by 500 μM glutamate, a concentration below the EC50 for GluK2, were also inhibited by 100 μM zinc (48% ± 10%, n = 10; data not shown).

9 In addition, Horowitz et al assessed 383 patients with no significant risk factor associated with hemorrhage to evaluate the clinical relevance of routine hemoglobin testing following an elective cesarean section. Their result showed

that the Hb concentration pre and post operation were 12.24 ± 1.09 and 10.87 ± 1.2 g/dl, respectively. They found no statistically significant difference among the patients according to indication and concluded that routine postoperative Hb measurement after an uncomplicated cesarean section in asymptomatic low-risk women is not necessary and should be eliminated.10 In another study, the evaluation of 421 cases with Modulators unplanned and uneventful low-risk women with no postoperative signs or symptoms for anemia by Api et al revealed selleck chemical that the mean pre and postoperative Hb levels were 11.7 ± 1.99 g/dl and 11.24 ± 1.99 g/dl, respectively (P < 0.001). Their results showed that there was a decrease http://www.selleckchem.com/products/KU-55933.html in Hb concentrations in 72% of the patients, whereas 24.5% experienced an increase and 3.5% showed no change, postoperatively. They suggest that routine Hb testing following uneventful, unplanned cesarean section neither changes postoperative management nor determines the patients requiring blood transfusion. 6 In the present study, we tried to find whether, is it necessary to carry out

pre operation blood typing and screening testing and post cesarean section Hb testing for low-risk women who underwent unplanned and uneventful operation. In our study, the mean preoperative hemoglobin was 12.4 ± 0.95 g/dl, whereas it was 11.8 ± 1.08 g/dl, postoperatively. Moreover, in our study, just two cases with parity over 4, showed Hb drop between 20 and 30% that could be due to previous injury of uterine, but none of them need to blood transfusion. Also, there was no relationship between maternal age, number of gestation, previous delivery, abortion and type of blood group with Hb decline

in our study. Performing blood typing and screening test before operation and Hb testing post operation in low-risk women who undergo unplanned from and uneventful cesarean section is unnecessary and can be eliminated. All authors have none to declare. “
“La dysfonction des cordes vocales (DCV), adduction inappropriée des cordes vocales classiquement pendant l’inspiration, est diagnostiquée à l’aide d’une laryngoscopie sus-glottique. Le diagnostic de DCV est difficile et mal codifié. “
“Selon le rapport de l’Organisation mondiale de la santé sur les facteurs de risque cardiovasculaire, l’hypertension artérielle (HTA) est responsable de 18 % des décès dans les pays riches et de 45 % des décès cardiovasculaires [1] et génère de lourds handicaps liés aux accidents vasculaires cérébraux (AVC), à la démence, à l’insuffisance cardiaque et à l’insuffisance rénale chronique. En 2008, les décès cardiovasculaires représentaient, en France, 30 % de l’ensemble des décès [2].

Recently, a new rotavirus vaccine, ROTAVAC®, based on the 116E rotavirus strain and manufactured by Bharat Biotech International Limited of India, demonstrated efficacy in a pivotal clinical trial in India [10] and [11]. Additional rotavirus vaccines are in inhibitors various stages of preclinical and clinical development. The parameters for the success of such trials from a regulatory perspective will likely differ from the parameters for policy or vaccine introduction decisions, and thus the various study designs used to evaluate efficacy in these trials

are likely to differ. To properly frame the results of clinical trials Trametinib nmr conducted with new vaccines, we reviewed the available literature on efficacy trials of rotavirus vaccines in low-resource settings in Africa and Asia. While acknowledging the importance of safety in regulatory and policy decisions, we limited this review to efficacy outcomes, and to the currently approved and recommended vaccines (Rotarix®, RotaTeq®). Both Rotarix® and RotaTeq® were already approved by

international regulatory authorities www.selleckchem.com/products/PD-0332991.html when tested in Africa and Asia, and thus those trials were conducted primarily to inform policy. Under the assumption that aspects of study design and population characteristics will influence the point estimates of efficacy obtained, we propose that comparisons of point estimates of efficacy from different trials may be challenging, and should be done with a clear understanding of trial design and the variables

that could influence such comparisons. Table 1 provides a number of factors that are known or hypothesized to influence rotavirus vaccine immunogenicity and/or efficacy, with references and examples from clinical trials. We then used these study design characteristics as a framework for evaluating the efficacy data from the new oral rotavirus vaccine, ROTAVAC® as an example of how to interpret appropriately new efficacy results (Table 2). Concomitant administration of oral poliovirus vaccines (OPV) with oral rotavirus vaccines reduces the immunogenicity of rotavirus vaccines, as measured by serum IgA antibody responses and rotavirus vaccine shedding, when compared Linifanib (ABT-869) with administration of the two vaccines separated in time by 1–2 weeks (Table 1) [12], [13] and [14]. This lower immunogenicity would be expected to result in no effect, or a reduction in efficacy, against clinical outcomes. In moderate to high resource settings, rotavirus vaccines were administered with inactivated poliovirus vaccines (IPV), or separated from OPV administration by at least 2 weeks. In trials performed to date in low resource settings, most of the children received OPV concomitantly with RVs as shown in Table 2. The exception was the trial of RotaTeq® in Africa, where only 35% of children received OPV with RV.

Although the percentage of GFP+ cells in the CD11clow/− population Libraries following 10 μg, 1 μg and 0.1 μg Ag doses appeared elevated compared to PBS/LPS control, particularly Metabolism inhibitor in draining CLN and BLN, these were not statistically significant. The proportion of CD11clow/− cells containing GFP

following 100 μg Ag, was higher in the local cervical and brachial LNs than in more distal inguinal and axial LNs (data not shown). Background correction, calculated by subtracting mean values for PBS control from dose values revealed that GFP+ cells could be detected at low Ag doses ( Fig. 2A and B, insets). The amount of cell-associated GFP from doses less than 100 μg may be below the level of sensitivity of GFP detection by flow cytometry. Lymphoid tissue autofluorescence also impacts on assay sensitivity. Analysis of cells displaying pMHC complexes (i.e. Y-Ae+) revealed that we could detect complexes in more than 20% of all CD11chigh cells in the draining CLNs (Fig. 2C) and BLNs (not shown) at the 100 μg dose. Decreasing amounts of Ag resulted in corresponding

decreases in the percentages of CD11c+Y-Ae+ cells, with the limit of detection of pMHC complexes between 1 μg and 100 ng of administered Ag. pMHC complex detection in CD 11clow/− ABT 737 cells showed a similar trend. As was the case for detection of GFP+ cells, variability within the small group (n = 3), limited statistical significance. Both the CD11chigh and CD11clow/− populations also showed increased, although not statistically significant, Y-Ae mean fluorescence down to a dose of 100–10 ng Ag (data not shown). These results indicate that with controlled and careful detailed analyses, we can detect both Ag and cells displaying pMHC complexes following administration these of about 1 μg–100 ng Ag, and this is the upper limit of Ag that we might expect to be produced following pDNA injection. The kinetics of Ag distribution and presentation is likely to vary depending on the route (e.g. subcutaneous vs. intramuscular) and the type of immunisation (e.g. protein vs. pDNA), and we wished to determine the kinetics of appearance of pMHC complexes for both protein and pDNA immunisation. The aim of this protein

injection study was to study the kinetics of Ag distribution in a widely studied situation such as subcutaneous injection. As has been shown for EαRFP previously [1], EαGFP+ cells, i.e. cell-associated EαGFP, can be found in the neck-draining CLNs and BLNs within 1 h of Ag injection in both CD11chigh (Fig. 3A) and CD11clow/− (Fig. 3B) cells. Fluorescence microscopy indicated that in addition to this cell-associated Ag, much of the injected Ag appeared to be extracellular (Fig. 1D). After this initial wave of antigen positive cells in the draining LNs, the number of cells carrying or associated with Ag decreased until 12–24 h when GFP+ cells reappeared in draining LNs. CD11c+GFP+ cells reappeared in the BLNs prior to their reappearance in the CLNs (Fig.