, 2009a and Conte et al., 2009b). However, such tissue disruption is comparable following injections of the same

amount of saline. Therefore, www.selleckchem.com/Androgen-Receptor.html the important issue is whether damage occurs in the transport zones. Based on saline injection controls (Figure S1) and histology, we found no evidence of damaged cells in the transport zones. Even after long survival times postinjection, we did not observe decays in MR enhancement, thus ruling out the possibility of secondary degeneration in these remote transport zones. Histological studies have shown that CTB is transported in both directions along axons, both retrogradely to cell bodies and anterogradely to presynaptic terminals (Luppi et al., 1986, Bruce and Grofova, 1992, Angelucci et al., 1996, Sakai et al., 1998, Sakai et al., 2000 and Wu and Kaas, 2000). Is our CTB-based compound also transported in both directions? The current results clarify half of this two-part question. As is typical in the brain, transport from S1 to VPL, and from S1 and Po, are known to be reciprocal. Thus in these areas, our MR enhancement

does not distinguish between the two directions of transport. However, connections between S1 and Rt and CPu are atypically unidirectional: S1 projects to both Rt and CPu, but neither Rt nor CPu projects back to S1 (Kaas and VEGFR inhibitor Ebner, 1998 and Liu and Jones, 1999. Gerfen, 1989, Kincaid and Wilson, 1996 and Hoover et al., 2003). In addition, it is known that sensory neurons in the olfactory epithelium project anterogradely to the OB. Thus we can conclude that the GdDOTA-CTB is many transported anterogradely, at least. Injections of manganese chloride, coupled with MR imaging (MEMRI), have also been widely used to map brain connections in vivo (Pautler et al., 1998, Saleem et al., 2002, Wu et al., 2006, Tucciarone et al., 2009 and Chuang and Koretsky, 2009). However, it is complicated to interpret the relationship of MEMRI data to the density of anatomical connections. First, MRI enhancement due to manganese reflects functional (i.e., calcium-related) activity (Lin and Koretsky, 1997, Aoki et al., 2002,

Aoki et al., 2004, Yu et al., 2005 and Eschenko et al., 2010a) as well as anatomical connections. Second, manganese may be released from tissue after uptake (unpublished observations); manganese at the injection sites spreads quickly and continuously (e.g., Figures 7 and S6, see also Tucciarone et al., 2009). Thus MEMRI has not been used to measure connections across very small distances, such as those across cortical laminae. Third, manganese is transported multisynaptically, not monosynaptically. In some experiments, this multisynaptic transport can be an advantage. However as described above, this can be a disadvantage if it is crucial to define each serial step of a given circuit. Moreover, diffusion of the manganese, coupled with the multisynaptic transport, could produce nonspecific transport.

Such segregation of AP-dependent and AP-independent presynaptic regulatory mechanisms may be particularly significant in circumstances where evoked release probability is low (Borst, 2010), and the two types of synaptic signals are difficult to differentiate. Under these circumstances, selective augmentation of spontaneous neurotransmitter release may facilitate neurotrophic, homeostatic PI3K inhibitor or other signaling functions of released neurotransmitter substances without compromising their function in AP-evoked information transfer. Recent studies suggest that the AP-independent forms of neurotransmitter release are critical

in the regulation of behavioral outcomes such as nociception, memory processing, and response to antidepressants (Andresen et al., 2012, Autry et al., 2011, Jin et al., 2012, Kavalali and Monteggia, 2012, Xu et al., 2012 and Nosyreva et al., 2013). This premise is consistent with the recent

behavioral analysis of VAMP7 knockout mice that revealed a deficit in anxiety-related behaviors (Danglot et al., 2012). In this way, identification of the vesicular release machineries and neuromodulators that specifically modify AP-independent forms of neurotransmission may provide novel avenues to manipulate neurotransmission without altering AP-dependent information processing. These types of approaches provide a promising strategy to uncover the functional roles FK228 of these unconventional forms of neuronal communication in the regulation of behavior in normal as well as in disease states. Dissociated hippocampal cultures were prepared from postnatal day 0–3 Sprague-Dawley rats of either sex

as described previously (Kavalali et al., 1999). For syb2 knockout (KO) and SNAP-25 KO experiments, dissociated hippocampal cultures were prepared from embryonic day 18 mice constitutively deficient in syb2 (syb2−/−) or SNAP-25 (SNAP-25−/−) as well as their littermate controls (Schoch et al., 2001 and Washbourne et al., 2002). ApoER2 KO and VLDLR KO cultures were prepared from mice generated by constitutive deletion of Apoer2 ( Trommsdorff et al., 1999) and vldlr genes ( Frykman et al., 1995). To generate below neurons deficient in p110α and p110β isoforms of PI3K (gift of Drs. Joel Elmquist, UT Southwestern, and Jean Zhao, Dana-Farber Cancer Institute), hippocampal cultures from mice expressing conditionals alleles of p110α and p110β genes were infected with lentivirus expressing Cre. Lentiviral expression system shows high infection efficacy (>90%) as previously demonstrated by full rescue of synaptic transmission in syb2−/− cultures by lentiviral expression of syb2 ( Deák et al., 2006). All experiments were performed on 14–21 days in vitro (DIV) cultures. All experiments were performed in accordance with protocols approved by the UT Southwestern Institutional Animal Care and Use Committee.

01, p < 0.001; category effect: F(2,17) = 175.27, p < 0.05; interaction between age and category: F(2,17) = 212.04, p < 0.05). Pairwise comparisons of L > F, L > U, and U > F were done on the hemodynamic responses in each category selective ROI to each stimulus (Table S2). Face-selective regions showed a statistically higher percent signal change to Face stimuli than to Learned or Untrained shapes, and all the Shape selective regions showed significantly higher signal change to Learned symbols and Untrained shapes

compared to Face stimuli. The Learned symbol region showed significantly higher signal change to Learned symbols compared to Untrained shapes and Faces, in juveniles but not in adults. To explore the difference between juveniles and adults in the responsiveness of the Learned symbol region we first defined an Average Learned symbol ROI by combining scans from all three juvenile monkeys and aligning them to SAR405838 datasheet a standard monkey template (McLaren et al., 2009). The average Learned symbol-selective ROI comprised 114 contiguous voxels that were preferentially more active in the combined juveniles data set to Learned symbols than to Untrained shapes or to Faces (p < 0.001 for both contrasts). We counted the voxels in all six individual monkeys within the average Learned symbol ROI that were selectively responsive to Learned symbols in each monkey (Table S3).

This average ROI contained significantly more voxels selectively responsive to Learned much click here symbols in juveniles (mean = 28) compared to adults (mean = 4); (t(10) = –3.17, p = 0.011, two-tailed t test). The fact that fMRI showed a Learned symbol-selective region in juveniles but not in adults could reflect the better performance of the juveniles compared to the adults, rather than a qualitative difference between the two groups. Therefore, to ask whether the Learned symbol region was exclusively present in juveniles, and not simply less active, or in a different place, in adult monkeys, we further calculated,

in each whole brain, the number of voxels that were significantly selective for Learned symbols, at three different thresholds (Table S4), without smoothing or clustering. Juvenile monkeys showed significantly more voxels selective for Learned symbols than adults did, irrespective of the threshold used, indicating that the juveniles showed qualitatively different responses to the Learned symbols (p < 0.01 at all thresholds tested). The novel functional specialization in juveniles for Learned symbols is probably not due to low-level differences between Learned symbols and Untrained shapes, such as degree of curvature or retinotopic representation, or to attentional differences, because we did not see any Learned symbol specialization in either of the adult-trained monkeys or in the naive adult.

To test for autonomy of effect, we also recorded from dMNs in the islet−/− mutant. Dorsal MNs do not express Selleck BMS354825 islet, and IKfast currents of WT and mutant larvae were statistically indistinguishable ( Figure 2C; WT 60.1 ± 4.3 pA/pF versus islet−/− 68.2 ± 5.9 pA/pF p = 0.28). We conclude that loss of islet only affects IKfast in vMNs in which it is normally expressed, but not in dMNs that lack

expression of this transcription factor. We further noted that loss of islet from the vMNs resulted in a transformation of IKfast to recapitulate the magnitude of this same current recorded in dMNs. When averaged responses of islet−/− vMNs and WT dMNs were superimposed, only small kinetic differences remain ( Figure 2D). Such an observation is entirely consistent

with, and indeed predictive of, the magnitude of IKfast being regulated by endogenous expression of Islet. Fast K+ currents in Drosophila neurons are encoded by one or more of at least three different genes: two voltage-gated fast-activating and inactivating channels (A-currents) termed Shal and Shaker (Sh) and a Ca2+-activated BK channel termed slowpoke ( Baker and Salkoff, 1990; Elkins et al., 1986; Singh and Wu, 1990). To determine which K+ current is increased in vMNs click here following loss of islet, we used specific blockers of these individual currents. We first explored whether IKslowpoke is repressed by Islet. To do so we added Cd2+ to the bath solution. Cd2+ blocks Ca2+ entry mafosfamide and, as a consequence, prevents activation of Ca2+-activated K+ channels. Addition of Cd2+ did not diminish the increase in IKfast observed in the vMNs in islet−/− mutants (data not shown). We conclude from this that Islet does not influence IKslowpoke.

By contrast, the presence of α-Dentrotoxin (DTx), a potent and specific blocker for Sh-mediated K+ currents (Ryglewski and Duch, 2009; Wu et al., 1989), completely abolishes the increase of IKfast seen in the vMNs in islet−/− ( Figure 3A; control 58.5 ± 6.9 versus DTx 43.1 ± 2.7 pA/pF p ≤ 0.05). Indeed, IKfast values obtained in the presence of DTx closely mirrored untreated WT vMNs (43.1 ± 2.7 versus 42.6 ± 3.1 p = 0.9). That DTx negates the islet−/− phenotype is consistent with Islet inhibiting a Sh-mediated K+ current in WT vMNs. To verify this prediction, we recorded IKfast in a Sh;islet double mutant. Similarly, under these conditions, peak current density of IKfast in the double mutant was indistinguishable from WT vMNs ( Figure 3A; p = 0.24). Our data are consistent with Islet acting to repress expression of Sh in vMNs. Moreover, removal of this repression results in expression of Sh-mediated K+ channels that confer “dorsal-like” electrical properties. This model posits, therefore, that dMNs normally express a Sh-mediated K+ current. To test this, we compared IKfast in dMNs between WT and in the presence of either DTx or in a Sh null mutant (Sh[14]).

09 mg mL−1 and LDT with LC90 = 0.03 mg mL−1) and L. sidoides (LDT with LC90 = 0.03 mg mL−1). These results were significant when compared to those observed for the other plant extracts evaluated against H. contortus. Spigelia anthelmia showed 100% inhibition in the EHT and 81.2% in the LDT at a concentration of 50 mg mL−1 ( Assis et al., 2003). Maciel et al. (2006) observed better efficacy of ethanol seed extract (100% inhibition at 1.56 mg mL−1) and leaf extract (98.24% at 12.5 mg mL−1) of Melia azedarach on eggs of H. contortus and the ethanol extract of leaves (91.64% at 50 mg mL−1) on the larvae of the same species. Macedo et al. (2010), in turn, analyzed the effect of Eucalyptus staigeriana essential oil and observed inhibitory

effect on eggs (99.27% at 1.35 mg mL−1) Z-VAD-FMK price and larvae (99.20% at 5.4 mg mL−1). Cocos nucifera showed selleck screening library 100% of inhibition at 5 mg mL−1 in the EHT and 99.77% at 80 mg mL−1 in the LDT ( Oliveira et al., 2009b). The potential larvicidal activity of the aqueous extract (97.3% inhibition at 150 mg mL−1) and ethanol extract (99.6% inhibition at 60 mg mL−1) of Anacardium humile leaves on gastrointestinal nematodes in sheep was reported by Nery et al. (2010). Some studies have analyzed the antiparasitic action of plant extracts on parasites of rats or mice and sheep. Among these, Borba and Amorim (2004) studied the action of

extracts of Chenopodium ambrosioides L. on the oxyurids Syphacia obvelata and SB-3CT Aspiculuris tetraptera, obtaining negative results for all concentrations tested. This same species was evaluated in rats against Strongyloides venezuelensis and showed efficacy in reducing the EPG (75.89%) and the

number of adult parasites (86.31%) at 400 mg kg−1 ( Bernardes, 2006). In field trials with sheep, Camurça-Vasconcelos et al. (2008) administered the essential oil of L. sidoides at concentrations of 230 and 283 mg kg−1 and observed a reduction in the EPG count in the evaluations conducted 7 and 14 days after the treatment: 38% and 30% and 45% and 54%, respectively. In our evaluation with rats, a significant reduction was observed in the number of adult parasites at both doses tested (150 and 250 mg kg−1) compared to the control group, treated with sorbitol. The highest mean number of EPG was recorded 7 or 8 days after infection of rats with S. venezuelensis, after which egg output showed progressive reduction. Therefore, the trend in EPG values observed in the sorbitol group reflects a natural reduction of the egg elimination. Due to its sedative effect, observed after the first administration of the oil, we chose not to perform the following two treatments to prevent the animals’ death. In vivo tests are needed to evaluate the effect of plant extracts with significant results in vitro on parasites. However, the possible toxic effects on the target hosts should be performed earlier. In conclusion, of the five plant extracts evaluated, M. piperita, P. tuberculatum and L. sidoides showed the best efficacy against H.

Subject-by-subject signal-to-noise values from each sensory experiment were correlated with signal-to-noise values from the other experiments (Figure 5) or with IQ/ADOS behavioral scores (Figure 5). A randomization test was used to assess the significance of each correlation value: a null distribution of 10,000 random correlation values was generated by randomly shuffling signal-to-noise values find protocol across individuals and statistical significance was defined as the 95th percentile of this distribution. Note that this is a more conservative statistical test than the Pearson’s

correlation coefficient, which assumes a normal distribution. We computed accuracy on the letter repetition-detection task by determining the fraction of trials where letter repeats were accurately reported from all possible letter repeats. Reaction time was measured from the appearance of the repeating letter to the button press (Figure S6). Two complementary analyses were carried out on the six estimated head motion parameters (three translations and three rotations) that were extracted from the Brainvoyager 3D

motion correction analysis. The standard deviation of head motion parameters and the mean frame-by-frame head motion were statistically indistinguishable across groups. Furthermore, projecting out head motion estimates from the fMRI data did not alter the findings (see Figure S7). Heart rate and respiration were measured Linifanib (ABT-869) using Siemens hardware and software, which automatically

identifies and marks time points containing heart beats and peaks PD0325901 ic50 of respiration. Physiology was sampled simultaneously with fMRI during a separate rest experiment, which was performed within the same scanning session as the sensory experiments. We computed heart and respiration rates and compared their average and temporal variability across groups (Figure S8). Eye position was acquired with an MRI compatible eye tracker (EyeTrac6, Applied Science Laboratories, Bedford, MA). Successful eye tracking was performed in six subjects with autism and three controls. We compared the average variance of the x and y eye position traces both throughout the entire experiment and also specifically within windows starting at stimulus onset and ending 500 after stimulus offset (Figure S8). This work was supported by Simons Foundation SFARI grant 177638 (D.J.H., M.B., and I.D.), ISF and Bikura grants (R.M.), Clore and Kahn postdoctoral fellowships (I.D.), Pennsylvania Department of Health SAP grant 4100047862 and NICHD/NIDCD PO1/U19 (M.B.). This research was also supported by the NIH/NICHD University of Pittsburgh Autism Center of Excellence HD055748. “
“Lens-based fluorescence microscopes, especially their confocal and two-photon variants (Denk et al.

While this approach might also be challenging in some cell types (GJs are dendrodendritic in most mammalian neurons), the M-cell and the

CEs offer several unusual anatomical and physiological characteristics Lumacaftor cost that make it possible to estimate these parameters in vivo: (1) CE afferents terminate with a single contact and are tightly segregated to the distal portion of the lateral dendrite of the M-cell; (2) the M-cell lateral dendrite as well as both the axons and terminals of CEs are accessible for intracellular recordings; and (3) the M-cell and the CEs have comparable and unusually fast membrane time constants, estimated to be 400 μs in the M-cell (Fukami et al., 1965) and 200 μs in CEs (Curti et al., 2008), which allow the use of physiological signals, such as action potentials, for measurements of CCs. Due to spatial considerations, measurements of CCs during simultaneous recordings of CE afferents in the VIIIth nerve root and the M-cell dendrite are useful to expose asymmetry of electrical transmission (Figures 4A and S3B) but not

accurate enough for estimating GJ conductance (see below). To overcome this problem, we calculated average values of CCs for the population of afferents, using values obtained under various experimental arrangements that maximize their accuracy (see below). The “population CC” in the orthodromic direction Thymidine kinase (CE to M-cell) for a number of CEs was estimated as the ratio between the average amplitude BMS-387032 in vitro of the electrical component (or coupling potential) of the unitary postsynaptic potential and the average amplitude of the presynaptic spike (CC,

postsynaptic coupling potential/presynaptic spike; Figure 4A). The orthodromic coupling potential (recorded during paired recordings with intradendritic recordings in the terminal field of CEs) averaged 0.73 ± 0.04 mV SEM (n = 76). (Because the strength of electrical synapses between individual CEs varies dramatically [Smith and Pereda, 2003], it was not possible to assign differences in the amplitude of individual coupling potentials to their relative position within the dendritic field and therefore correct for potential electrotonic attenuation. Thus, although potentially slightly underestimated, we believe the average amplitude of orthodromic coupling potentials represents the most appropriate value to use for calculating the CC in the orthodromic direction.) During simultaneous recordings, the amplitude of the presynaptic spike evoked at the recording site with long (200 ms) depolarizing pulses does not represent the spike that ultimately generates coupling, as the spike recorded at the site of depolarization regenerates in subsequent nodes and, finally, at the presynaptic terminal (see Figure S4).

The ratio of apoptotic cells was significantly increased, dependent on PPD concentration (i.e., >20 μM, consistent with the above cell proliferative data), compared with control (Fig. 4A; P < 0.01). HCT-116 and SW-480 cells were treated with different concentrations (15, 20, 25, 30, and 35 μM) of PPD for 48 h and the cell cycle was examined by flow cytometry. As shown in Fig. 4B, PPD-induced G1 cell cycle arrest in a concentration-dependent manner in both cell lines (both P < 0.01). HCT-116 cells were selected to perform mRNAs expression profiling analysis on six samples, find more including three control vehicle treated cells and different concentrations and time points of PPD-treated cells.

We first performed an unsupervised, two-way (genes against samples), hierarchical cluster analysis (HCA). Remarkably, three PPD-treated cell samples (24p20, 48p20, 48p25) clearly grouped into one cluster, while three normal control cell samples also grouped together and formed a cluster (Fig. 5A). 204 genes significantly changed (over 1.5-fold) after PPD treatment. A sub-analysis based 79 genes significantly altered (over 2-fold) (Fig. 5B). 20 of the most upregulated and downregulated genes were compiled based on the microarray data, shown in Table 1 and Table 2. Among the genes that were selleck chemicals significantly altered when treated

with PPD in HCT-116 cells, six downregulated genes (CLSPN, CCNA2, SPAG5, DNM3, DHCR24, DSCC1) and five upregulated genes (BTG1, DDIT4, PDCD4, KLF4, NRP1) were validated by quantitative real-time RT-PCR. The same RNA samples for microarray were used to generate cDNA templates for reverse transcription reactions. The SYBR green-based real-time RT-PCR analysis was then carried out. Consistent much with the microarray data, the 11 selected genes showed the same expression profile as the microarray data presented (Fig. 5C and D). We performed gene network analysis using the 204 significant genes from our microarray analysis through the Ingenuity Pathway Analysis (IPA). A bar plot presenting ten classic

pathways related to tumorigenesis is shown in Fig. 6A. Among them, apoptosis, proliferation, and angiogenesis were significantly induced. This is consistent with our in vitro data, suggesting that PPD is probably involved in cancer cell growth by modulating these processes. The selected regulatory cell death pathway gene network is shown in Fig. 6B, in which 23 affected genes of this network were either upregulated or downregulated after PPD treatment. Among the genes, DR4 and DR5 are important members of the tumor necrosis factors (TNF) family. It appears that HCT-116 cell apoptosis was induced after PPD exposure by the interaction of p53 and DR4/DR5, and suggests that the TRAIL pathway was associated with the PPD activities. CRC is one of the most common cancers worldwide (18).

In diagnostic research, a stepwise evaluation of tests is increasingly proposed considering not only the test’s technical reliability and accuracy but also its place in the clinical pathway and, eventually, its impact on patient outcomes (Van den Bruel et al 2007). Investigating the role and position

of measurements of passive movements of the extremities within clinical pathways for diagnosing disorders forms an unexplored field of research in physiotherapy and could improve the external validity of future reliability studies. With respect to internal validity, only two studies (Cibere et al 2004, Watkins et al 1991) satisfied all three criteria, suggesting unbiased estimates of inter-rater reliability. This disappointing finding is similar to those of reviews of measurements KRX-0401 nmr of upper extremity movements (Van de Pol et al 2010) and spinal movement (Seffinger et al 2004, Van Trijffel Ku 0059436 et al 2005). However, in many cases, these validity criteria could not be scored due to inadequate reporting of the

study protocol. In these cases, it was not possible to provide any indication of the presence and/or direction of the risk of bias. The criteria related to the stability of test circumstances, for both participants and raters, indicate underestimation of reliability if they are not met. Instability of the participants’ characteristics under study – in this case the joint’s mobility – may be caused by changes in the biomechanical properties of joint connective tissues as a result of natural variation over time or mobilising effects of the assessment procedure itself (Rothstein and Echternach 1993). Similarly, instability of the raters’ capability of making judgments may be the result of, for example, mental fatigue. A lack of appropriate blinding of raters, on the other hand, could lead to overestimation of reliability. all If several of these methodological

flaws are present, the direction of risk of bias is difficult to predict. Researchers should give careful consideration to ensuring stability of participants’ and raters’ characteristics during research and to provide detailed information on the study protocol by following the STARD statement (Bossuyt et al 2003a, Bossuyt et al 2003b). Similar recommendations for improving the reporting of reliability studies were made in the field of medical research (Gow et al 2008). A lack of inter-rater reliability adversely affects the accuracy of diagnostic decisions and subsequent treatment selection (Quinn 1989). This is particularly problematic when effective treatments are available and certain patients run the risk of not receiving them due to error and variation in decision-making among therapists. For instance, hip osteoarthritis is usually defined according to the clinical criteria of the American College of Rheumatology which include criteria about restrictions of physiological range of hip flexion and internal rotation (Altman et al 1991).

The total duration click here of each training trial was 500 s. A training trial started with placing the mouse in a square chamber with grid floor (Coulbourn Instruments; H10-11RTC, 120W × 100D × 120H). At 198 s, 278 s, 358 s, and 438 s, a foot shock was delivered (2 s, 0.70 mA). On days 2 and 3, FC+EXT mice were subjected to four extinction trials per day (day 2: E1 to E4; day 3: E5 to E8). Each extinction trial lasted 1,800 s with a trial interval of 2 hr. For each extinction trial, mice were placed in the same box used for

fear conditioning without receiving foot shocks. On day 4, FC and FC+EXT mice were tested over 500 s during a single retrieval test. For the retrieval test, mice were placed in the same box used for fear conditioning without receiving foot shocks. The design of experiment 2 is summarized in Figure 5A. The FC group in experiment 2 was subjected to the same protocol as the FC group in experiment 1. The HC group consisted of mice that stayed in their home cage during the entire experiment. HC mice were perfused at the same time as the FC mice. Freezing behavior was measured using a digital camera connected to a computer with Actimetrics FreezeFrame software. The bout length was 1 s and the threshold for freezing behavior was determined by an experimenter blind to

experimental conditions and animal group. Freezing scores were obtained by averaging freezing during minutes 2 and 3 of each trial. Ninety minutes after retrieval testing, mice were deeply anesthetized with ketamine/xylazine Birinapant molecular weight and transcardially perfused with 0.1 M phosphate buffer (PB) followed by 4% paraformaldehyde (PFA 4%) dissolved in 0.1 M PB. Brains were extracted and postfixed in PFA 4% for 24 hr. Brains were transferred to 30% sucrose for 48–72 hr before slicing 20 μm coronal sections of the entire brain using a cryostat. Sections were stored

in cryoprotectant at −20°C until use. Free-floating sections were rinsed extensively in PBS with 0.25% Triton X-100 (PBS-T). Sections Terminal deoxynucleotidyl transferase were blocked for 1 hr at room temperature in PBS-T with 10% normal goat serum (or 3% donkey serum for CB1R). Sections were incubated in rabbit anti-Zif268 (Santa-Cruz; polyclonal; 1:3,000) with either mouse anti-GAD67 (Millipore; monoclonal; 1:10,000), mouse anti-PV (Millipore; monoclonal; 1:2,000), mouse anti-CCK/Gastrin (Center for Ulcer Research and Education UCLA; monoclonal; 1:1,000), or goat anti-CB1 (kind gift of Dr. K. Mackie; polyclonal; 1:2,000) (Harkany et al., 2005). Additional primary antibodies used were rabbit anti-CamKII (kind gift of Dr. M. Jacob; polyclonal; 1:2,000) and rabbit anti-Rab3b (kind gift of Dr. T. Südhof; polyclonal; 1:4,000). Primary antibodies were diluted in the blocking solution, incubated at 4°C for 72 hr, and rinsed three times for 15 min in PBS-T.