Bisphenol A Exposure Induces Sensory Processing Deficits in Larval Zebrafish during Neurodevelopment.

Because of their ex utero development, relatively simple nervous system, translucency, and availability of tools to investigate neural function, larval zebrafish are an exceptional model for understanding neurodevelopmental disorders and the consequences of environmental toxins. Furthermore, early in development, zebrafish larvae easily absorb chemicals from water, a significant advantage over methods required to expose developing organisms to chemical agents in utero Bisphenol A (BPA) and BPA analogs are ubiquitous environmental toxins with known molecular consequences. All humans have measurable quantities of BPA in their bodies. Most concerning, the level of BPA exposure is correlated with neurodevelopmental difficulties in people. Given the importance of understanding the health-related effects of this common toxin, we have exploited the experimental advantages of the larval zebrafish model system to investigate the behavioral and anatomic effects of BPA exposure. We discovered that BPA exposure early in development leads to deficits in the processing of sensory information, as indicated by BPA's effects on prepulse inhibition (PPI) and short-term habituation (STH) of the C-start reflex. We observed no changes in locomotion, thigmotaxis, and repetitive behaviors (circling). Despite changes in sensory processing, we detected no regional or whole-brain volume changes. Our results show that early BPA exposure can induce sensory processing deficits, as revealed by alterations in simple behaviors that are mediated by a well-defined neural circuit.

The importance of air quality for underground spaces: An international survey of public attitudes.

Space is a resource that is constantly being depleted, especially in mega-cities. Underground workspaces (UGS) are increasingly being included in urban plans and have emerged as an essential component of vertical cities. While progress had been made on the engineering aspects associated with the development of high-quality UGS, public attitudes toward UGS as work environments (ie, the public's design concerns with UGS) are relatively unknown. Here, we present the first large-scale study examining preferences and attitudes toward UGS, surveying close to 2000 participants from four cities in three continents (Singapore, Shanghai, London, and Montreal). Contrary to previous beliefs, air quality (and not lack of windows) is the major concern of prospective occupants. Windows, temperature, and lighting emerged as additional important building performance aspects for UGS. Early adopters (ie, individuals more willing to accept UGS and thus more likely to be the first occupants) across all cities prioritized air quality. Present results suggest that (perceived) air quality is a key building performance aspect for UGS that needs to be communicated to prospective occupants as this will improve their attitudes and views toward UGS. This study highlights the importance of indoor air quality for the public.

Induction of Short-Term Sensitization by an Aversive Chemical Stimulus in Zebrafish Larvae.

Larval zebrafish possess a number of molecular and genetic advantages for rigorous biological analyses of learning and memory. These advantages have motivated the search for novel forms of memory in these animals that can be exploited for understanding the cellular and molecular bases of vertebrate memory formation and consolidation. Here, we report a new form of behavioral sensitization in zebrafish larvae that is elicited by an aversive chemical stimulus [allyl isothiocyanate (AITC)] and that persists for ≥30 min. This form of sensitization is expressed as enhanced locomotion and thigmotaxis, as well as elevated heart rate. To characterize the neural basis of this nonassociative memory, we used transgenic zebrafish expressing the fluorescent calcium indicator GCaMP6 (Chen et al., 2013); because of the transparency of larval zebrafish, we could optically monitor neural activity in the brain of intact transgenic zebrafish before and after the induction of sensitization. We found a distinct brain area, previously linked to locomotion, that exhibited persistently enhanced neural activity following washout of AITC; this enhanced neural activity correlated with the behavioral sensitization. These results establish a novel form of memory in larval zebrafish and begin to unravel the neural basis of this memory.

Health Effects of Underground Workspaces cohort: study design and baseline characteristics.

The development of underground workspaces is a strategic effort towards healthy urban growth in cities with ever-increasing land scarcity. Despite the growth in underground workspaces, there is limited information regarding the impact of this environment on workers' health. The Health Effects of Underground Workspaces (HEUW) study is a cohort study that was set up to examine the health effects of working in underground workspaces. In this paper, we describe the rationale for the study, study design, data collection, and baseline characteristics of participants. The HEUW study recruited 464 participants at baseline, of whom 424 (91.4%) were followed-up at 3 months and 334 (72.0%) at 12 months from baseline. We used standardized and validated questionnaires to collect information on socio-demographic and lifestyle characteristics, medical history, family history of chronic diseases, sleep quality, health-related quality of life, chronotype, psychological distress, occupational factors, and comfort levels with indoor environmental quality parameters. Clinical and anthropometric parameters including blood pressure, spirometry, height, weight, and waist and hip circumference were also measured. Biochemical tests of participants' blood and urine samples were conducted to measure levels of glucose, lipids, and melatonin. We also conducted objective measurements of individuals' workplace environment, assessing air quality, light intensity, temperature, thermal comfort, and bacterial and fungal counts. The findings this study will help to identify modifiable lifestyle and environmental parameters that are negatively affecting workers' health. The findings may be used to guide the development of more health-promoting workspaces that attempt to negate any potential deleterious health effects from working in underground workspaces.

Rapid habituation of a touch-induced escape response in Zebrafish (Danio rerio) Larvae.

Zebrafish larvae have several biological features that make them useful for cellular investigations of the mechanisms underlying learning and memory. Of particular interest in this regard is a rapid escape, or startle, reflex possessed by zebrafish larvae; this reflex, the C-start, is mediated by a relatively simple neuronal circuit and exhibits habituation, a non-associative form of learning. Here we demonstrate a rapid form of habituation of the C-start to touch that resembles the previously reported rapid habituation induced by auditory or vibrational stimuli. We also show that touch-induced habituation exhibits input specificity. This work sets the stage for in vivo optical investigations of the cellular sites of plasticity that mediate habituation of the C-start in the larval zebrafish.

Assessing the suitability of virtual reality for psychological testing.

Virtual reality (VR) is rapidly becoming an inexpensive, mainstream technology. VR technology is superambulatory as it allows participants to be examined under standardized environments and tests anywhere. In addition, it can test participants in different virtual spaces, including environments that are unsafe, inaccessible, costly or difficult to set up, or even nonexistent. We summarize the benefits and potential problems of VR technology, but we also move beyond theoretical approaches and present a customizable, open-source VR system (PSY-VR) that allows scalable psychological testing in modifiable VR environments. This system allows users to modify the environment using a simple graphical interface, without programming expertise. Moreover, as a proof-of-concept, we compare responses in a typical Flanker task between a real laboratory and a painstakingly matched virtual laboratory. Results indicate that the VR responses are comparable to real life testing, demonstrating the utility of VR for psychological assessment studies. The predicted rapid advancement of VR immersive technologies, as well the ease of their integration with physiological metrics ensures that VR-based assessment will be the modus operandi of psychological assessment in the future. This will allow controllable, low-cost assessment on a global scale. (PsycINFO Database Record (c) 2019 APA, all rights reserved).

A Psychosocial Approach to Understanding Underground Spaces.

With a growing need for usable land in urban areas, subterranean development has been gaining attention. While construction of large underground complexes is not a new concept, our understanding of various socio-cultural aspects of staying underground is still at a premature stage. With projected emergence of underground built environments, future populations may spend much more of their working, transit, and recreational time in underground spaces. Therefore, it is essential to understand the challenges and advantages that such environments have to improve the future welfare of users of underground spaces. The current paper discusses various psycho-social aspects of underground spaces, the impact they can have on the culture shared among the occupants, and possible solutions to overcome some of these challenges.

Role of protein synthesis and DNA methylation in the consolidation and maintenance of long-term memory in Aplysia.

Previously, we reported that long-term memory (LTM) in Aplysia can be reinstated by truncated (partial) training following its disruption by reconsolidation blockade and inhibition of PKM (Chen et al., 2014). Here, we report that LTM can be induced by partial training after disruption of original consolidation by protein synthesis inhibition (PSI) begun shortly after training. But when PSI occurs during training, partial training cannot subsequently establish LTM. Furthermore, we find that inhibition of DNA methyltransferase (DNMT), whether during training or shortly afterwards, blocks consolidation of LTM and prevents its subsequent induction by truncated training; moreover, later inhibition of DNMT eliminates consolidated LTM. Thus, the consolidation of LTM depends on two functionally distinct phases of protein synthesis: an early phase that appears to prime LTM; and a later phase whose successful completion is necessary for the normal expression of LTM. Both the consolidation and maintenance of LTM depend on DNA methylation.

Long-term habituation of the C-start escape response in zebrafish larvae.

The cellular and molecular basis of long-term memory in vertebrates remains poorly understood. Knowledge regarding long-term memory has been impeded by the enormous complexity of the vertebrate brain, particularly the mammalian brain, as well as by the relative complexity of the behavioral alterations examined in most studies of long-term memory in vertebrates. Here, we demonstrate a long-term form of nonassociative learning-specifically, long-term habituation (LTH)-of a simple reflexive escape response, the C-start, in zebrafish larvae. The C-start is triggered by the activation of one of a pair of giant neurons in the zebrafish's hindbrain, the Mauthner cells. We show that LTH of the C-start requires the activity of NMDA receptors and involves macromolecular synthesis. We further show that the long-term habituated reflex can by rapidly dishabituated by a brief tactile stimulus. Our results set the stage for rigorous, mechanistic investigations of the long-term memory for habituation of a reflexive behavioral response, one that is mediated by a relatively simple, neurobiologically tractable, neural circuit. Moreover, the demonstration of NMDAR and transcriptionally dependent LTH in a translucent vertebrate organism should facilitate the use of optical recording, and optogenetic manipulation, of neuronal activity to elucidate the cellular basis of a long-term vertebrate memory.

Processing of Phonemic Consonant Length: Semantic and Fragment Priming Evidence from Bengali.

Six cross-modal lexical decision tasks with priming probed listeners' processing of the geminate-singleton contrast in Bengali, where duration alone leads to phonemic contrast ([pata] 'leaf' vs. [pat:a] 'whereabouts'), in order to investigate the phonological representation of consonantal duration in the lexicon. Four form-priming experiments (auditory fragment primes and visual targets) were designed to investigate listeners' sensitivity to segments of conflicting duration. Each prime derived from a real word ([k(h)[symbol: see text]m]/[g(h)en:]) was matched with a mispronunciation of the opposite duration (*[k(h)[symbol: see text]m:]/*[g(h)en]) and both were used to prime the full words [k(h)[symbol: see text]ma] ('forgiveness') and [g(h)en:a] ('disgust') respectively. Although all fragments led to priming, the results showed an asymmetric pattern. The fragments of words with singletons mispronounced as geminates led to equal priming, while those with geminates mispronounced as singletons showed a difference. The priming effect of the real-word geminate fragment was significantly greater than that of its corresponding nonword singleton fragment. In two subsequent semantic priming tasks with full-word primes a stronger asymmetry was found: nonword geminates (*[k(h)[symbol: see text]m:a]) primed semantically related words ([marjona] 'forgiveness') but singleton nonword primes (*[ghena]) did not show priming. This overall asymmetry in the tolerance of geminate nonwords in place of singleton words is attributed to a representational mismatch and points towards a moraic representation of duration. While geminates require a mora which cannot be derived from singleton input, the additional information in geminate nonwords does not create a similar mismatch.

Reinstatement of long-term memory following erasure of its behavioral and synaptic expression in Aplysia.

Long-term memory (LTM) is believed to be stored in the brain as changes in synaptic connections. Here, we show that LTM storage and synaptic change can be dissociated. Cocultures of Aplysia sensory and motor neurons were trained with spaced pulses of serotonin, which induces long-term facilitation. Serotonin (5HT) triggered growth of new presynaptic varicosities, a synaptic mechanism of long-term sensitization. Following 5HT training, two antimnemonic treatments-reconsolidation blockade and inhibition of PKM--caused the number of presynaptic varicosities to revert to the original, pretraining value. Surprisingly, the final synaptic structure was not achieved by targeted retraction of the 5HT-induced varicosities but, rather, by an apparently arbitrary retraction of both 5HT-induced and original synapses. In addition, we find evidence that the LTM for sensitization persists covertly after its apparent elimination by the same antimnemonic treatments that erase learning-related synaptic growth. These results challenge the idea that stable synapses store long-term memories.

Asymmetric processing of durational differences - electrophysiological investigations in Bengali.

Duration is used contrastively in many languages to distinguish word meaning (e.g. in Bengali, [pata] 'leaf' vs. [pat:a] 'whereabouts'). While there is a large body of research on other contrasts in speech perception (e.g. vowel contrasts and consonantal place features), little work has been done on how durational information is used in speech processing. In non-linguistic studies of low-level processing, such as visual and non-linguistic acoustic pop-out tasks, an asymmetry is found where additional information is more readily detected than missing information. In this study, event-related potentials were recorded during two cross-modal auditory-visual semantic priming studies, where nonword mispronunciations of spoken prime words were created by changing the duration of a medial consonant (real word [dana] 'seed'>nonword [dan:a]). N400 amplitudes showed an opposite asymmetric pattern of results, where increases in consonantal duration were tolerated and led to priming of the visual target, but decreases in consonantal duration were not accepted. This asymmetrical pattern of acceptability is attributed to the fact that a longer consonant includes all essential information for the recognition of the original word with a short medial consonant (a possible default category) and any additional information can be ignored. However, when a consonant is shortened, it lacks the required durational information to activate the word with the original long consonant.

Learning and memory in zebrafish larvae.

Larval zebrafish possess several experimental advantages for investigating the molecular and neural bases of learning and memory. Despite this, neuroscientists have only recently begun to use these animals to study memory. However, in a relatively short period of time a number of forms of learning have been described in zebrafish larvae, and significant progress has been made toward their understanding. Here we provide a comprehensive review of this progress; we also describe several promising new experimental technologies currently being used in larval zebrafish that are likely to contribute major insights into the processes that underlie learning and memory.

Presynaptic NMDA receptor mechanisms for enhancing spontaneous neurotransmitter release.

NMDA receptors (NMDARs) are required for experience-driven plasticity during formative periods of brain development and are critical for neurotransmission throughout postnatal life. Most NMDAR functions have been ascribed to postsynaptic sites of action, but there is now an appreciation that presynaptic NMDARs (preNMDARs) can modulate neurotransmitter release in many brain regions, including the neocortex. Despite these advances, the cellular mechanisms by which preNMDARs can affect neurotransmitter release are largely unknown. Here we interrogated preNMDAR functions pharmacologically to determine how these receptors promote spontaneous neurotransmitter release in mouse primary visual cortex. Our results provide three new insights into the mechanisms by which preNMDARs can function. First, preNMDARs can enhance spontaneous neurotransmitter release tonically with minimal extracellular Ca(2+) or with major sources of intracellular Ca(2+) blocked. Second, lowering extracellular Na(+) levels reduces the contribution of preNMDARs to spontaneous transmitter release significantly. Third, preNMDAR enhance transmitter release in part through protein kinase C signaling. These data demonstrate that preNMDARs can act through novel pathways to promote neurotransmitter release in the absence of action potentials.

Synaptic dysfunction and abnormal behaviors in mice lacking major isoforms of Shank3.

SHANK3 is a synaptic scaffolding protein enriched in the postsynaptic density (PSD) of excitatory synapses. Small microdeletions and point mutations in SHANK3 have been identified in a small subgroup of individuals with autism spectrum disorder (ASD) and intellectual disability. SHANK3 also plays a key role in the chromosome 22q13.3 microdeletion syndrome (Phelan-McDermid syndrome), which includes ASD and cognitive dysfunction as major clinical features. To evaluate the role of Shank3 in vivo, we disrupted major isoforms of the gene in mice by deleting exons 4-9. Isoform-specific Shank3(e4-9) homozygous mutant mice display abnormal social behaviors, communication patterns, repetitive behaviors and learning and memory. Shank3(e4-9) male mice display more severe impairments than females in motor coordination. Shank3(e4-9) mice have reduced levels of Homer1b/c, GKAP and GluA1 at the PSD, and show attenuated activity-dependent redistribution of GluA1-containing AMPA receptors. Subtle morphological alterations in dendritic spines are also observed. Although synaptic transmission is normal in CA1 hippocampus, long-term potentiation is deficient in Shank3(e4-9) mice. We conclude that loss of major Shank3 species produces biochemical, cellular and morphological changes, leading to behavioral abnormalities in mice that bear similarities to human ASD patients with SHANK3 mutations.

NR3A-containing NMDARs promote neurotransmitter release and spike timing-dependent plasticity.

Recent evidence suggests that presynaptic-acting NMDA receptors (preNMDARs) are important for neocortical synaptic transmission and plasticity. We found that unique properties of the NR3A subunit enable preNMDARs to enhance spontaneous and evoked glutamate release and that NR3A is required for spike timing-dependent long-term depression in the juvenile mouse visual cortex. In the mature cortex, NR2B-containing preNMDARs enhanced neurotransmission in the absence of magnesium, indicating that presynaptic NMDARs may function under depolarizing conditions throughout life. Our findings indicate that NR3A relieves preNMDARs from the dual-activation requirement of ligand-binding and depolarization; the developmental removal of NR3A limits preNMDAR functionality by restoring this associative property.

Habituation of the C-start response in larval zebrafish exhibits several distinct phases and sensitivity to NMDA receptor blockade.

The zebrafish larva has been a valuable model system for genetic and molecular studies of development. More recently, biologists have begun to exploit the surprisingly rich behavioral repertoire of zebrafish larvae to investigate behavior. One prominent behavior exhibited by zebrafish early in development is a rapid escape reflex (the C-start). This reflex is mediated by a relatively simple neural circuit, and is therefore an attractive model behavior for neurobiological investigations of simple forms of learning and memory. Here, we describe two forms of short-lived habituation of the C-start in response to brief pulses of auditory stimuli. A rapid form, persisting for ≥1 min but <15 min, was induced by 120 pulses delivered at 0.5-2.0 Hz. A more extended form (termed "short-term habituation" here), which persisted for ≥25 min but <1 h, was induced by spaced training. The spaced training consisted of 10 blocks of auditory pulses delivered at 1 Hz (5 min interblock interval, 900 pulses per block). We found that these two temporally distinguishable forms of habituation are mediated by different cellular mechanisms. The short-term form depends on activation of N-methyl-d-aspartate receptors (NMDARs), whereas the rapid form does not.

Influence of the NR3A subunit on NMDA receptor functions.

Various combinations of subunits assemble to form the NMDA-type glutamate receptor (NMDAR), generating diversity in its functions. Here we review roles of the unique NMDAR subunit, NR3A, which acts in a dominant-negative manner to suppress receptor activity. NR3A-containing NMDARs display striking regional and temporal expression specificity, and, unlike most other NMDAR subtypes, they have a low conductance, are only modestly permeable to Ca(2+), and pass current at hyperpolarized potentials in the presence of magnesium. While glutamate activates triheteromeric NMDARs composed of NR1/NR2/NR3A subunits, glycine is sufficient to activate diheteromeric NR1/NR3A-containing receptors. NR3A dysfunction may contribute to neurological disorders involving NMDARs, and the subunit offers an attractive therapeutic target given its distinct pharmacological and structural properties.

Downregulation of NR3A-containing NMDARs is required for synapse maturation and memory consolidation.

NR3A is the only NMDA receptor (NMDAR) subunit that downregulates sharply prior to the onset of sensitive periods for plasticity, yet the functional importance of this transient expression remains unknown. To investigate whether removal/replacement of juvenile NR3A-containing NMDARs is involved in experience-driven synapse maturation, we used a reversible transgenic system that prolonged NR3A expression in the forebrain. We found that removal of NR3A is required to develop strong NMDAR currents, full expression of long-term synaptic plasticity, a mature synaptic organization characterized by more synapses and larger postsynaptic densities, and the ability to form long-term memories. Deficits associated with prolonged NR3A were reversible, as late-onset suppression of transgene expression rescued both synaptic and memory impairments. Our results suggest that NR3A behaves as a molecular brake to prevent the premature strengthening and stabilization of excitatory synapses and that NR3A removal might thereby initiate critical stages of synapse maturation during early postnatal neural development.