, 1968, Koltzenburg et al., 1997 and Lynn and Carpenter, 1982). Hairy skin LTMRs are physically and functionally associated with hair follicles, and in these species hair follicles fall into three distinct types according to length, thickness, and presence of kinks selleck chemicals in the hair shaft (Schlake, 2007) (Figure 1B). Hairy skin is innervated by several LTMR subtypes that fall into distinctive Aβ-, Aδ-, and C-type categories depending on conduction velocities. We are beginning to appreciate the morphological and molecular diversity of hair follicle afferents and their intricate patterns of connections with different hair follicle types (Bourane et al.,

2009, Li et al., 2011, Luo et al., 2009, Millard and Woolf, 1988 and Wu et al., 2012). Indeed, a new picture has emerged, in which hairy skin is a highly specialized sensory organ, as or more complex than glabrous skin, with each hair follicle type representing its own unique mechanosensory unit. Aβ-LTMRs. The first category of low-threshold mechanosensors in hairy skin check details fall into the

Aβ category of conduction velocities. As for glabrous skin, hair follicle-innervating Aβ-LTMRs are divided into two groups according to their firing adaptation rates: slowly adapting (SA) and RA LTMRs. Hairy skin SAI-LTMRs are associated with the Merkel cell complex, or touch dome, found within the epidermal/dermal junction first surrounding the mouths of Guard hairs of rodents (Figure 1B) and their firing properties are similar to those recorded from SAI-LTMRs of glabrous skin (Woodbury and Koerber, 2007). SAII response properties have also been identified in rodent hairy skin, but, as already discussed, the

anatomical correlate of SAII units remains controversial (Wellnitz et al., 2010 and Zimmermann et al., 2009). The most well-characterized hairy skin physiological responses that fall under the category of Aβ/myelinated afferents are the Aβ RA-LTMRs. Historically, the physiological properties of hairy skin RA-LTMRs have been classified by responses to movement of individual hair follicle types at a controlled speed and direction (Brown and Iggo, 1967 and Burgess et al., 1974). Across species, hairy skin RA-LTMRs share some basic physiological characteristics. First, hairy skin RA-LTMRs are not spontaneously active nor do they respond to thermal stimuli. Second, their responses to hair follicle movement can exhibit either few action potentials or a stream of action potentials proportional to velocity and final amplitude of displacement. Third, their physiological receptive field sizes vary extensively across the body, with a trend toward a decrease in receptive field size in the most distal sections of body hair, i.e., extremities. Aβ RA-LTMR responses in hairy skin arise from longitudinal lanceolate endings that surround hair follicles.

The above examples of heterogeneity in CSC populations, as well as several others [55] are likely to reflect plasticity in the CSC phenotype. Additional plasticity is also reflected in studies that show that non-CSCs can acquire CSC properties [56] and [57]. For example, similar to normal stem cells, a microenvironmental niche has been shown to be required to maintain glioma and skin cancer CSCs [58] and [59], and this is probably also the case for other tumor types [60]. A perivascular location can actually Small Molecule Compound Library be the driving force that leads to the acquisition of CSC properties

by non-CSC subpopulations [61]. Thus extrinsic microenvironmental cues are emerging learn more as important determinants of the CSC population. Metastases can occur many years after surgical removal of the primary tumor, which has given rise to the concept of dormancy. These late-developing metastases are thought to develop from DTCs that have become re-activated after remaining in a stable dormant state over a prolonged period [62]. For example, after radical prostatectomy for prostate cancer, almost half of all patients have detectable DTCs in

their bone marrow more than 5 years after their surgery [63]. Dormant tumor cells can exist in a quiescent state, or as micrometastases in which proliferation is balanced by cell death through apoptosis [7]. Reactivation of these dormant cells can be due to changes in the tumor cells themselves, for example due to loss of metastasis suppressor genes that regulate dormancy [64], as well as to modification of their microenvironment,

for example extracellular matrix second (ECM) remodeling and recruitment of inflammatory cells [65] and [66]. The activation of the growth of indolent tumor cells by bone marrow-derived cells (BMDC) recruited in response to osteopontin produced by a second remote “instigator” tumor may also reflect the re-animation of dormant cells [67]. Due to their quiescence or slow turnover, dormant tumor cells are resistant to conventional cytotoxic therapies because their intrinsic quiescence makes them insensitive to DNA-damaging agents that specifically target cycling cells [68]. An elegant recent study that looked at the mechanism behind the re-activation of dormant breast cancer cells in the bone marrow provides evidence that intrinsic changes in gene expression in tumor cells can relieve dormancy [41]. Metastases growing out in the bone marrow after long latency periods were found to express VCAM-1, in contrast to the parental clone that was originally injected into the experimental animals. In further rounds of injection into animals, these VCAM-1-expressing cells were able to form bone metastases without entering dormancy.

, 2010). There are some common themes for channel biogenesis shared by tetrameric VGICs and the pentameric LGICs. Surface expression of nicotinic acetylcholine receptors and GABAA receptors depends on the evolutionarily conserved ER membrane complex (EMC) learn more that regulates protein folding and ER-associated degradation (Richard et al., 2013). Unlike dimers lacking GABAA receptor α or β subunit that are retained in the ER, assembly of heterodimers

of α and β subunits involves calnexin and the immunoglobulin heavy chain binding protein BiP (Bradley et al., 2008, Connolly et al., 1996 and Luscher et al., 2011). In addition to ER chaperones such as BiP/GRP78, calnexin, and ERp57 (Blount and Merlie, 1991, Colombo et al., 2013, Gelman et al., 1995, Paulson et al., 1991 and Wanamaker and Green, 2007), the ER membrane protein RIC-3 regulates acetylcholine

receptor assembly and ER dwell time (Alexander et al., 2010). One striking finding is that, often, interaction with small molecules, including the natural ligand of a channel, can influence biogenesis. Not only does glutamate act as a chemical chaperone in the biogenesis of glutamate receptors (Penn and Greger, 2009), but GABA may be an intracellular chaperone for GABAA receptor biogenesis (Eshaq et al., 2010) and nicotine may act in a similar way for nascent α4β2 and α3β4 nicotinic acetylcholine receptors in SB203580 the ER to enhance their surface expression (Colombo et al., 2013, not Govind et al., 2012, Mazzo et al., 2013, Miwa et al., 2011, Sallette et al., 2005 and Srinivasan et al., 2011). Similar mechanisms may be involved in the rescue of deficient trafficking of a mutant HERG potassium channel in human long QT syndrome

by HERG channel blockers (Rajamani et al., 2002 and Zhou et al., 1999), and the ability of sulfonylureas to function as chemical chaperones to rescue the trafficking defects of ATP-sensitive potassium channels bearing certain mutations that cause congenital hyperinsulinism (Yan et al., 2004). Together, these observations suggest that a better understanding of ion channel biogenesis should enlighten understanding of basic issues about membrane protein folding and may also yield new means to intervene in cases in which channel activity has gone wrong in disease states. Ensuring that only properly folded and assembled channels make it to plasma membrane is important as channels that lack key elements of regulation could cause serious dysfunction. Starting with ER quality control, proteins that reside in the ER or Golgi shuttle channel complexes between these intracellular compartments before mature channels proceed in forward traffic to reach the cell membrane (Colombo et al., 2013, Dancourt and Barlowe, 2010, Deutsch, 2003, Luscher et al., 2011 and Schwappach, 2008).

However, the mechanisms by which macrophages

kill Leishmania in dogs have not been investigated as thoroughly ( Rodrigues et al., 2007). The immune response against Leishmania sp. is highly dependent on the microbicidal action of macrophages, which are actually the host cell target of this protozoan; however, they have full capacity for antigen presentation and establishment C59 wnt solubility dmso of an effective response against the parasite ( Pinelli et al., 1999). Thus, to develop new approaches for analyzing the immune response of naturally L. chagasi-infected dogs or dogs immunized against CVL, in vitro co-culture systems with macrophages and purified T-lymphocytes would be useful. However, there is so far no standardized methodology for this purpose, and these tests usually only involve a system

with peripheral blood mononuclear cells (PBMCs) without purified T-lymphocyte subsets ( Holzmuller et al., 2005, Rodrigues et al., 2007 and Rodrigues et al., 2009). The development of additional methodologies for evaluating the immune system in veterinary medicine, especially in experimental dog models, is required. Such an advance would contribute to the identification of biomarkers related to interactions between innate and adaptive immune responses of dogs. In this context, we aimed to further analyze the immune response by using standardized methodologies for a co-culture system of canine L. chagasi-infected either macrophages and for obtaining purified CD4+ and CD8+ T cells. This approach could contribute to identifying specific immune response biomarkers for developing a resistance or susceptibility profile in CVL, which ABT888 could be used in both vaccine and treatment strategies against the parasite. Healthy mongrel dogs, both sexes with a mean age of 7 months,

born and raised in a kennel at the Center of Animal Science, Federal University of Ouro Preto, were used in the experiments of (i) establishment of in vitro conditions of monocytes differentiated into macrophages infected with L. chagasi (n = 5) and (ii) purification procedures of T-cell subsets (CD4+ and CD8+) using microbeads (n = 12). The animals received all the appropriate health management before entering the experiment, having received anti-helmintic treatment (plus Chemital®, Chemitec Agro-Veterinary LTDA., BRA) and vaccination against rabies (Tecpar, BRA), distemper, adenovirus type 2, coronavirus, parainfluenza, parvovirus, and Leptospira (HTLP 5/CV-L Vanguard®, Pfizer, BRA). The study protocol was approved by the Ethical Committee for the Use of Experimental Animals of the Universidade Federal de Ouro Preto, Ouro Preto – MG, Brazil. This study used a wild-type strain of L. chagasi (C46) isolated from an infected dog of Governador Valadares, MG, and previously characterized in hamsters ( Moreira et al., 2012). This strain was grown in culture medium NNN/LIT (Sigma Chemical Co.

, 2001 and Zou et al., 2008). The Crb family proteins are single-pass type I transmembrane proteins, and their intracellular domains function to assemble other components of the Crb complex. Aside from their functions with respect to polarity maintenance, genetic studies in Drosophila have shown that Crb inhibits Notch signaling ( Herranz et al., 2006 and Richardson and Pichaud, 2010). Nevertheless,

it remains to be uncovered how Crb BKM120 nmr interacts with Notch for the regulation of neurogenesis. Notch receptors are large, single-pass, type I transmembrane proteins that maintain neuroepithelial cells in the undifferentiated state in a transcription-dependent manner (Louvi and Artavanis-Tsakonas, 2006). The binding of Notch ligands,

such as Delta, triggers the proteolytic cleavage of Notch by multiple proteases, including γ-secretase, which results in the release of the Notch intracellular domain (NICD). NICD is translocated to the nucleus where it forms a transcriptional complex with Mastermind and a member of the CBF1/RBP-J, Su(H), Lag1 (CSL) family; thereafter, it promotes the expression of genes that inhibit the differentiation of neuroepithelial selleck chemicals cells (Louvi and Artavanis-Tsakonas, 2006). In addition to this well-characterized canonical Notch pathway, it has been recently reported that NICD activates a small GTPase R-Ras that

facilitates the cellular adhesion of CHO cells in a transcription-independent manner (Hodkinson et al., 2007). However, it is not known how this noncanonical Notch pathway participates in vertebrate neural development. In the present study, we demonstrate that Crb binds to the extracellular domain of Notch and inhibits its activation, and that a component of the Crb complex, Mosaic eyes [Erythrocyte membrane protein band Resminostat 4.1-like 5 (Epb41l5) according to the Zebrafish Nomenclature Committee; known as Yurt in Drosophila, and Lulu1 or YMO1 in mammals and hereafter referred to as Moe] counteracts this inhibition. Furthermore, we show that the Crb⋅Moe complex-Notch signaling also maintains neuroepithelial apicobasal polarity via the R-Ras-dependent noncanonical Notch pathway. Therefore, our results suggest that the Crb⋅Moe complex-Notch signaling plays pivotal roles both in the restriction of neuroepithelial mitosis in the apical area and in the maintenance of apicobasal polarity of neuroepithelial cells. Tg(CM-isl1:GFP)rw0 (hereinafter referred to as isl1:GFP) transgenic zebrafish express the GFP in most of their cranial motor neurons, including the vagus motor neurons ( Higashijima et al., 2000).

Accordingly, the four sectors covering the lesion in SM’s RH were centered on the posterior tip of the right lateral fusiform gyrus in each

subject. Figure 5B shows the position of the grid in control subject C1. Posterior and ventral sectors of the grid covered parts of VO1/2, while dorsal sectors covered most parts of functionally localized LOC, which was defined on the basis of anatomical and functional characteristics. As in previous studies (e.g., Malach, et al., 1995), LOC was defined as a contiguous cluster localized near the lateral occipital sulcus that responded more strongly to the presentations of intact pictures of objects versus their scrambled counterparts (p < 0.0001). LOC was separately defined for each fMRI study. For example, 2D objects were contrasted with scrambled Selleck PF-06463922 2D objects (Figures 2A and 2B). For the functional Selleckchem AZD9291 analysis of grid sectors, the four sectors encompassing the lesion site were excluded. It is important to note that the grid analysis does not assume or require corresponding functional grid locations across subjects, since we probed general response characteristics such as visual responsiveness, object-related and -selective responses, which are typical for this portion of cortex. The visual responsiveness of cortex in the penumbra of the lesion was investigated by contrasting activations evoked by presentations of all types of objects versus blank images (Figure 5C; Table S2). Figure S4 shows the activations

evoked by presentations of individual types of objects versus blank images. The criterion for significant activation in a given grid-sector was defined as an activated volume of at least 50% of the grid sector’s volume, that is 108 mm3, or 4 voxels (p < 0.001) for all subsequent analyses. To exclude the possibility that an arbitrary voxel threshold distorted the results, we performed a second analysis with a more lenient voxel threshold of 81 mm3, or 3 voxels (Figure S5), which yielded similar over results compared to the more conservative analysis presented here. In the controls, 79% ± 11% of the grid sectors in the

RH showed activation indicating that cortex covered by the grid responded well to visual stimulation. Similarly, 77% of the grid-sectors in the RH of control subject C1 showed visual activation. The sectors that were not visually responsive were located in anterior and ventral sectors of the grid. Eccentricity maps from the control subjects suggested that these locations represent the periphery as opposed to the fovea of the visual field (Arcaro et al., 2009). Thus, the lack of activation in these regions is likely due to the parafoveal location of the stimuli. In SM, 64% of the sectors in the RH showed activation. Interestingly, most sectors immediately surrounding the lesion were activated and sectors that were not responsive to visual stimulation, as in the control subjects, were located in anterior and ventral sectors of the grid.

Together, these data imply that mGluR-induced OPHN1 mediates LTD by promoting see more the internalization of AMPARs. Further support for these results, and mechanistic insight into how OPHN1 induction could regulate AMPAR endocytosis during mGluR-LTD, were provided by our finding that OPHN1 interacts with N-BAR domain-containing Endo2/3 core components of the postsynaptic clathrin-dependent endocytic machinery (Chowdhury et al., 2006). Interestingly, our data show that mGluR stimulation enhances OPHN1 association with Endo2/3 in a protein synthesis

dependent manner. And importantly, disruption of the OPHN1-Endo2/3 interaction impedes both mGluR-elicited persistent decreases in surface AMPARs and LTD. Notably, these effects are not attributable to some general disruption of AMPARs or the machinery that controls their trafficking, because disruption of the OPHN1-Endo2/3 interaction does not affect basal AMPAR levels or basal synaptic function. Thus, these data imply that the downregulation of surface AMPARs during mGluR-LTD requires OPHN1 induction and its ability to bind Endo2/3. Likely, OPHN1 induced upon mGluR activation, via the regulation of Endo2/3′s activities, increases the rate of AMPAR endocytosis. While our data demonstrate a requirement for

OPHN1 synthesis in mGluR-LTD, previous studies have shown that newly synthesized BMS-907351 price Arc protein is also required for this process (Waung et al., 2008), implying that both mGluR-induced OPHN1 and Arc, and perhaps other proteins, such as MAP1B and

STEP (Davidkova and Carroll, 2007 and Zhang et al., 2008), are likely to contribute jointly to LTD, and, moreover, that mGluR1/5 must coordinate the various translational control mechanisms involved. Of particular interest is that Arc also interacts with Endo2/3 and this interaction is important for the role of Arc in AMPAR trafficking (Chowdhury et al., 2006). Of note, OPHN1 and Arc interact with different regions of Endo2/3, with OPHN1 binding to the SH3 domain of Endo2/3, and Arc to the C terminus of the N-BAR domain of Endo2/3 (Chowdhury et al., 2006). Oxygenase Therefore, it is possible that newly synthesized OPHN1 and Arc cooperate at the level of Endo2/3 to promote mGluR-driven AMPAR endocytosis, either by regulating distinct aspects of Endo2/3 function or by promoting/engaging a common mechanism, at least under wild-type conditions. Importantly, a different mode of mGluR-LTD regulation seems to occur upon loss of FMRP. Indeed, previous studies demonstrated that mGluR-LTD in Fmr1 KO mice is distinctly different from that in wild-type mice. For instance, whereas mGluR-LTD in wild-type mice is protein synthesis dependent, it persists in the absence of protein synthesis in Fmr1 KO mice ( Hou et al., 2006 and Nosyreva and Huber, 2006).

The membranes were labeled with 1.5 mol% DiO (3,3′-dioctadecyloxacarbocyanine; Invitrogen). The GUVs were formed by the drying rehydration procedure, as described in van den Bogaart et al. (2011). Briefly, 1 mg/ml total lipid concentration in methanol was mixed with 1.5 mol% dioleoyl-PiP3 (1,2-dioleoyl-sn-glycero-3-[phosphoinositol-3′,4’,5′-trisphosphate];

Avanti Polar Lipids) in a 1:2:0.8 volume mixture of chloroform, methanol, and water. Subsequently, 3 mol% of Atto647N-syntaxin-1A (residues 257-288; Atto647N from Atto-Tec) in 2,2,2-trifluoroethanol (TFE) was added to the lipid mixture. We then dried 1 μl on www.selleckchem.com/products/i-bet151-gsk1210151a.html a microscope coverslip for 2 min at 50°C–60°C, followed by rehydration in 20 mM HEPES (pH 7.4). GUVs were imaged using a confocal microscope. Competitive binding experiments

were performed as described in Murray and Tamm (2009) by recording emission spectra of 100-nm-sized liposomes composed of a 4:1 molar ratio of DOPC/DOPS and prepared by extrusion through 100 nm polycarbonate membranes as described in van den Bogaart et al. (2007), with a 1:5,000 molar protein-to-lipid ratio of Atto647N-labeled Syntaxin1A (residues 257–288) and 1:5,000 buy Ku-0059436 of bodipy-labeled PI(4,5)P2 (bodipy-TMR-PI(4,5)P2,C16; Echelon Biosciences). No additional lipid was added or 1:5,000 or 1:500 of unlabeled PI(4,5)P2 or 1:5,000 of unlabeled PI(3,4,5)P2 was added. Excitation was at 544 nm and the excitation and emission slit widths were 1 nm and 5 nm, respectively. A spectrum in the presence of 0.05% Triton X-100 was recorded to correct for the fluorescence crosstalk (gray). Immunohistochemistry was performed as described in Kasprowicz et al. (2008), except for Syntaxin1A labeling; larval fillets were fixed for 15 min in Bouin’s fixative and fixed larvae were blocked with 0.25% BSA and 5% NGS in PBS. Antibodies used were the following: Ms anti-FasII1D4 1:20 (Vactor et al., 1993),

Ms anti-DLG4F3 1:250 (Parnas et al., 2001), Ms anti-CSP6D6 1:50 PAK6 (Zinsmaier et al., 1994), Ms anti-BRPNC82 1:100 (Wagh et al., 2006), Ms anti-Syntaxin8C3 1:20 (Schulze and Bellen, 1996) (Developmental Hybridoma Studies Bank), Rb anti-Dap160 1:200 (Roos and Kelly, 1998), Rb anti-Endo 1:200 (Verstreken et al., 2002), anti-HA 1:200, and Rb anti-RBP 1:500 (Liu et al., 2011). GFP or Venus was not visualized with antibodies but their fluorescence was imaged directly. Images were captured on a Zeiss 510 META or Leica DM 6000CS confocal microscope with a 63× NA 1.4 oil lens. Labeling intensity in single section confocal images was quantified as the mean gray value of boutonic fluorescence corrected for background in the muscle; all quantifications were performed on confocal images. Intensity line plots were generated by quantifying boutonic circumference fluorescence intensity in ImageJ and plotting the intensity values versus the normalized bouton circumference.

Volumes were typically 200 μm × 200 μm × 45 μm. Laser power exiting the objective ranged from 12–60 mW and was continuously adjusted depending on instantaneous focal depth. GCaMP3 was excited at 960 nm and emission was collected

with a green 2″ filter (542 nm center; 50 nm band; Semrock) via GaAsP photomultiplier tubes (Hamamatsu). Neurons were confirmed to be within a particular cortical area by comparison of two-photon images of surface vasculature above the imaging site with surface vasculature from widefield (intrinsic autofluorescence signal) retinotopic mapping. Recording sessions were 3–5 hr in duration. Viral expression of GCaMP3 permitted recording from neurons across multiples cortical areas in the same mice on different days (Andermann et al., 2010, Dombeck et al., 2010, Mank et al., PLX4032 mouse find more 2008, O’Connor et al., 2010 and Tian et al., 2009). When recording from the same cortical region on multiple days, previously imaged neurons were relocated and an adjacent volume was selected to ensure that all neurons in the sample were unique. During imaging, mice were placed on a 6″ foam trackball (Plasteel) that could spin noiselessly on ball bearings (McMaster-Carr).

We monitored trackball revolutions using a custom photodetector circuit. In a subset of experiments, we recorded eye movements using a CMOS camera (Mightex; 20 Hz) and infrared illumination Resminostat (720–900 nm bandpass filters, Edmund). To achieve accurate stimulation at temporal frequencies of 0.5–24 Hz, we used a 120 Hz LCD monitor (Samsung 2233RZ, 22″) calibrated (at each stimulus frequency) using

a spectrophotometer (Photoresearch PR-650; see also Wang and Nikolić, 2011). Waveforms were also confirmed to be sinusoidal by measuring luminance fluctuations of a full-field sinusoidally modulated stimulus (using a photomultiplier tube, Hamamatsu). The monitor was positioned so that the stimulus patch was 21 cm from the contralateral eye. Stimuli were centered at monocular locations of 70° to 115° eccentricity and −5° to 14° elevation (which provided maximal separation of responsive regions across visual cortical areas, Figure 1A). For cellular imaging, local 40° Gabor-like circular patches (sigmoidal 10%–90% falloff in 10°) containing sine-wave drifting gratings (80% contrast) were presented for 5 s, followed by 5 s of uniform mean luminance (46 cd/m2). In the spatial frequency × temporal frequency protocol (Figure 2), we presented upward-drifting gratings at 5 spatial frequencies (0.02, 0.04, 0.08, 0.16, and 0.32 cycles per degree, cpd) and 7 temporal frequencies (0.5, 1, 2, 4, 8, 15, and 24 Hz) for a total of 35 stimulus types plus 10% blank trials. In the spatial frequency × direction protocol (Figure 5), we presented up to 6 spatial frequencies (0.02, 0.04, 0.08, 0.16, and 0.

Since repetitive stimulation can release endocannabinoid 2-arachidonoyl

glycerol from MSNs (Kano et al., 2009; Maejima et al., 2005), we applied a cannabinoid CB1 receptor antagonist to block the endocannabinoid-mediated retrograde suppression of corticostriatal synapses and to evaluate unadulterated presynaptic function. In wild-type and PCDH17−/− mice at P21–P23, the normalized Selleckchem Vemurafenib EPSC amplitude decreased gradually during prolonged repetitive stimulation, but the depression of the normalized EPSC amplitude was significantly weaker in PCDH17−/− mice ( Figure 6D). There was no significant difference in average amplitudes of the first ten EPSCs between wild-type and PCDH17−/− mice ( Figure S6A). Since the numbers of total SVs was increased in PCDH17−/− synapses at three weeks of age ( Figures 5A and 5B), the weaker synaptic depression following repetitive stimulation VEGFR inhibitor is thought to result from the increased number of vesicles available for release during enhanced activity. In contrast, the extent of synaptic depression was similar in wild-type and PCDH17−/− mice at P16–P18 ( Figure S6C), suggesting that PCDH17’s regulation

of synaptic depression is developmental stage-dependent. Taken together, these electrophysiological data suggest that overall synaptic transmission efficacy is enhanced at anterior corticostriatal excitatory synapses in PCDH17−/− mice. To examine behavioral abnormalities in PCDH17−/− mice, we performed a battery of behavioral tests for evaluating sensory and motor functions, cognition, anxiety, and depression. We employed the tail suspension test and the forced swim test, which are widely used for assessing antidepressant-like activity in mice. In both tests, PCDH17−/− mice were less immobile than wild-type Phosphatidylinositol diacylglycerol-lyase mice ( Figures 7A and 7B), suggesting

that PCDH17−/− mice were less susceptible to depression than wild-type mice. Alternatively, the reduced immobility of PCDH17−/− mice in these tests might have been caused by an increase in spontaneous activity ( Cryan and Holmes, 2005). To check this possibility, we performed the open field test to measure spontaneous locomotor activity of PCDH17−/− mice. The results showed that there were no significant differences between wild-type and PCDH17−/− mice in terms of immobility time, total distance traveled, or the amount of rearing activity observed during the test ( Figure 7C), suggesting that spontaneous activity was normal in PCDH17−/− mice. As alteration in fearfulness can also affect performance in tests for depression ( Cryan and Holmes, 2005), we also analyzed anxiety-related behavior.