Remodeling of Purinergic Receptor-Mediated Ca2+ Signaling as a Consequence of EGF-Induced Epithelial-Mesenchymal Transition in Breast Cancer Cells

Background The microenvironment plays a pivotal role in tumor cell proliferation, survival and migration. Invasive cancer cells face a new set of environmental challenges as they breach the basement membrane and colonize distant organs during the process of metastasis. Phenotypic switching, such as that which occurs during epithelial-mesenchymal transition (EMT), may be associated with a remodeling of cell surface receptors and thus altered responses to signals from the tumor microenvironment. Methodology/Principal Findings We assessed changes in intracellular Ca 2+ in cells loaded with Fluo-4 AM using a fluorometric imaging plate reader (FLIPR TETRA) and observed significant changes in the potency of ATP (EC 50 0.175 μM (-EGF) versus 1.731 μM (+EGF), P<0.05), and the nature of the ATP-induced Ca 2+ transient, corresponding with a 10-fold increase in the mesenchymal marker vimentin (P<0.05). We observed no change in the sensitivity to PAR2-mediated Ca 2+ signaling, indicating that these alterations are not simply a consequence of changes in global Ca 2+ homeostasis. To determine whether changes in ATP-mediated Ca 2+ signaling are preceded by alterations in the transcriptional profile of purinergic receptors, we analyzed the expression of a panel of P2X ionotropic and P2Y metabotropic purinergic receptors using real-time RT-PCR and found significant and specific alterations in the suite of ATP-activated purinergic receptors during EGF-induced EMT in breast cancer cells. Our studies are the first to show that P2X 5 ionotropic receptors are enriched in the mesenchymal phenotype and that silencing of P2X 5 leads to a significant reduction (25%, P<0.05) in EGF-induced vimentin protein expression. Conclusions The acquisition of a new suite of cell surface purinergic receptors is a feature of EGF-mediated EMT in MDA-MB-468 breast cancer cells. Such changes may impart advantageous phenotypic traits and represent a novel mechanism for the targeting of cancer metastasis

Face recognition is similarly affected by viewpoint in school-aged children and adults

There is an ongoing debate on the question when face processing abilities mature. One aspect that has been part of this debate is the ability to recognize faces in and across different viewpoints. Here, we tested 128 participants consisting of school-age children (ages, 5 –10 years) and adults (ages, 19–37 years) in two experiments to investigate the effects of different viewpoints (including front, three-quarter, profile view) on face recognition during development. Furthermore, we compared recognition performance for faces to that of another object category (cars). In the first experiment ($\it n$ = 88) we tested if the pattern of performance for faces presented in different viewpoints is similar in school-aged children and adults. Participants completed a two-alternative- forced-choice (2AFC) memory task comprising images of both faces and cars in front, three-quarter and profile view, which were presented in the same viewpoint during learning and testing. In the second experiment ($\it n$ = 40) we tested if face recognition is similarly affected by viewpoint changes in children and adults. In this experiment the 2AFC memory task included a change of viewpoint between learning and testing. While in both experiments we found higher recognition performance for faces with increasing age, the overall pattern of both viewpoint and viewpoint-change-effects and also the difference between view-change- and no-change-conditions was similar across age groups. In contrast to faces, no viewpoint effects were observed in cars (experiment 1), viewpoint change effects, however, were similar for cars and faces (experiment 2). In sum, our results suggest early maturity of the ability to recognize faces in and across different viewpoints

Face recognition deficits in autism spectrum disorders are both domain specific and process specific

Although many studies have reported face identity recognition deficits in autism spectrum disorders (ASD), two fundamental question remains: 1) Is this deficit "process specific" for face memory in particular, or does it extend to perceptual discrimination of faces as well? And 2) Is the deficit "domain specific" for faces, or is it found more generally for other social or even nonsocial stimuli? The answers to these questions are important both for understanding the nature of autism and its developmental etiology, and for understanding the functional architecture of face processing in the typical brain. Here we show that children with ASD are impaired (compared to age and IQ-matched typical children) in face memory, but not face perception, demonstrating process specificity. Further, we find no deficit for either memory or perception of places or cars, indicating domain specificity. Importantly, we further showed deficits in both the perception and memory of bodies, suggesting that the relevant domain of deficit may be social rather than specifically facial. These results provide a more precise characterization of the cognitive phenotype of autism and further indicate a functional dissociation between face memory and face perception

Looking tasks online

Online experimentation is emerging as a new methodology within classical data acquisition in psychology. It allows for easy, fast, broad, and cheap data conduction from the comfort of people’s homes. To add another method to the array of available tools, here we used recent developments in web technology to investigate the technical feasibility of online HyperText Markup Language-5/JavaScript-based video data recording. We employed a preferential looking task with children between 4 and 24 months. Parents and their children participated from home through a three-stage process: First, interested adults registered and took pictures through a webcam-based photo application. In the second step, we edited the pictures and integrated them into the design. Lastly, participants returned to the website and the video data acquisition took place through their webcam. In sum, we were able to create and employ the video recording application with participants as young as 4 months old. Quality-wise, no participant had to be removed due to the framerate or quality of videos and only 7% of data was excluded due to behavioral factors (lack of concentration). Results-wise, interrater reliability of rated looking side (left/right) showed a high agreement between raters, Fleiss’ Kappa, $\kappa$ = 0.97, which can be translated to sufficient data quality for further analyses. With regard to on-/off-screen attention attribution, we found that children lost interest after about 10 s after trial onset using a static image presentation or 60 s total experimental time. Taken together, we were able to show that online video data recording is possible and viable for developmental psychology and beyond

A new 3-day standardized eyeblink conditioning protocol to assess extinction learning from infancy to adulthood

Associative learning can be observed from the neonatal period onward, providing opportunities to examine changes in basic learning and memory abilities. One method that is suitable to study associative learning is classical eyeblink conditioning (EBC) which is dependent on the cerebellum. Extinction learning can be systematically investigated in this paradigm by varying the context during learning and extinction. Because of methodological difficulties and ethical challenges, no studies have compared extinction learning using EBC across human development. Our goal was to test feasibility of a 3-day delay EBC paradigm that can be used from infancy to adulthood. Acceptance/safety was tested especially for infancy by investigating attrition rates and parental report on infant wellbeing. On a paradigm side, we tested if the paradigm leads to successful acquisition and extinction. An air puff served as unconditional stimulus (US) and a tone as conditional stimulus (CS). On day 1 during acquisition, participants received 36 US–CS pairings in context A. On day 2, participants received 12 acquisition trials in context A to consolidate association learning, followed by 48 extinction trials (tone alone presentations) in context B. Renewal was assessed on day 3 and incorporated 12 CS alone trials presented in both the acquisition context and the extinction context. Eyeblink responses were videotaped and coded offline. The protocol was tested with 12–36-months-old infants ($\it N$ = 72), adolescents ($\it N$ = 8), and adults ($\it N$ = 8). Concerning the acceptance/safety side, attrition ranged from 21 to 58% in infant samples due to the complex preparation of the children for the paradigm. However, attrition is equal to or lower than other infant learning paradigms. Parents of infant samples were very interested in the paradigm and reported low levels of infant stress, exhaustion, and negative feelings during the sessions. Data quality was very high, and no participant had to be excluded because of insufficient data. Concerning the paradigm side, participants showed successful acquisition and extinction as a group. The procedure is ethically sound, feasible, tolerated by many infants, and acceptable among parents. The data show successful acquisition and extinction rates, making the paradigm a valuable tool for investigating developmental changes in extinction learning over the lifespan

U can touch this

New technological devices, particularly those with touch screens, have become virtually omnipresent over the last decade. Practically from birth, children are now surrounded by smart phones and tablets. Despite being our constant companions, little is known about whether these tools can be used not only for entertainment, but also to collect reliable scientific data. Tablets may prove particularly useful for collecting behavioral data from those children (1–10 years), who are, for the most part, too old for studies based on looking times and too young for classical psychophysical testing. Here, we analyzed data from six studies that utilized touch screen tablets to deliver experimental paradigms in developmental psychology. In studies 1 and 2, we employed a simple sorting and recall task with children from the ages of 2–8. Study 3 (ages 9 and 10) extended these tasks by increasing the difficulty of the stimuli and adding a staircase-based perception task. A visual search paradigm was used in study 4 (ages 2–5), while 1- to 3-year-olds were presented with an extinction learning task in study 5. In study 6, we used a simple visuo-spatial paradigm to obtain more details about the distribution of reaction times on touch screens over all ages. We collected data from adult participants in each study as well, for comparison purposes. We analyzed these data sets in regard to four metrics: self-reported tablet usage, completeness of data, accuracy of responses and response times. In sum, we found that children from the age of two onwards are very capable of interacting with tablets, are able to understand the respective tasks and are able to use tablets to register their answers accordingly. Results from all studies reiterated the advantages of data collection through tablets: ease of use, high portability, low-cost, and highlevels of engagement for children. We illustrate the great potential of conducting psychological studies in young children using tablets, and also discuss both methodologica lchallenges and their potential solutions