Developmental trajectory and sex differences in auditory processing in a PTEN-deletion model of autism spectrum disorders.

Autism Spectrum Disorders (ASD) encompass a wide array of debilitating symptoms, including severe sensory deficits and abnormal language development. Sensory deficits early in development may lead to broader symptomatology in adolescents and adults. The mechanistic links between ASD risk genes, sensory processing and language impairment are unclear. There is also a sex bias in ASD diagnosis and symptomatology. The current study aims to identify the developmental trajectory and genotype- and sex-dependent differences in auditory sensitivity and temporal processing in a Pten-deletion (phosphatase and tensin homolog missing on chromosome 10) mouse model of ASD. Auditory temporal processing is crucial for speech recognition and language development and deficits will cause language impairments. However, very little is known about the development of temporal processing in ASD animal models, and if there are sex differences. To address this major gap, we recorded epidural electroencephalography (EEG) signals from the frontal (FC) and auditory (AC) cortex in developing and adult Nse-cre PTEN mice, in which Pten is deleted in specific cortical layers (layers III-V) (PTEN conditional knock-out (cKO). We quantified resting EEG spectral power distribution, auditory event related potentials (ERP) and temporal processing from awake and freely moving male and female mice. Temporal processing is measured using a gap-in-noise-ASSR (auditory steady state response) stimulus paradigm. The experimental manipulation of gap duration and modulation depth allows us to measure cortical entrainment to rapid gaps in sounds. Temporal processing was quantified using inter-trial phase clustering (ITPC) values that account for phase consistency across trials. The results show genotype differences in resting power distribution in PTEN cKO mice throughout development. Male and female cKO mice have significantly increased beta power but decreased high frequency oscillations in the AC and FC. Both male and female PTEN cKO mice show diminished ITPC in their gap-ASSR responses in the AC and FC compared to control mice. Overall, deficits become more prominent in adult (p60) mice, with cKO mice having significantly increased sound evoked power and decreased ITPC compared to controls. While both male and female cKO mice demonstrated severe temporal processing deficits across development, female cKO mice showed increased hypersensitivity compared to males, reflected as increased N1 and P2 amplitudes. These data identify a number of novel sensory processing deficits in a PTEN-ASD mouse model that are present from an early age. Abnormal temporal processing and hypersensitive responses may contribute to abnormal development of language function in ASD.

Sex differences during development in cortical temporal processing and event related potentials in wild-type and fragile X syndrome model mice.

BACKGROUND: Autism spectrum disorder (ASD) is currently diagnosed in approximately 1 in 44 children in the United States, based on a wide array of symptoms, including sensory dysfunction and abnormal language development. Boys are diagnosed ~ 3.8 times more frequently than girls. Auditory temporal processing is crucial for speech recognition and language development. Abnormal development of temporal processing may account for ASD language impairments. Sex differences in the development of temporal processing may underlie the differences in language outcomes in male and female children with ASD. To understand mechanisms of potential sex differences in temporal processing requires a preclinical model. However, there are no studies that have addressed sex differences in temporal processing across development in any animal model of ASD. METHODS: To fill this major gap, we compared the development of auditory temporal processing in male and female wildtype (WT) and Fmr1 knock-out (KO) mice, a model of Fragile X Syndrome (FXS), a leading genetic cause of ASD-associated behaviors. Using epidural screw electrodes, we recorded auditory event related potentials (ERP) and auditory temporal processing with a gap-in-noise auditory steady state response (ASSR) paradigm at young (postnatal (p)21 and p30) and adult (p60) ages from both auditory and frontal cortices of awake, freely moving mice. RESULTS: The results show that ERP amplitudes were enhanced in both sexes of Fmr1 KO mice across development compared to WT counterparts, with greater enhancement in adult female than adult male KO mice. Gap-ASSR deficits were seen in the frontal, but not auditory, cortex in early development (p21) in female KO mice. Unlike male KO mice, female KO mice show WT-like temporal processing at p30. There were no temporal processing deficits in the adult mice of both sexes. CONCLUSIONS: These results show a sex difference in the developmental trajectories of temporal processing and hypersensitive responses in Fmr1 KO mice. Male KO mice show slower maturation of temporal processing than females. Female KO mice show stronger hypersensitive responses than males later in development. The differences in maturation rates of temporal processing and hypersensitive responses during various critical periods of development may lead to sex differences in language function, arousal and anxiety in FXS.

Developmental delays in cortical auditory temporal processing in a mouse model of Fragile X syndrome.

BACKGROUND: Autism spectrum disorders (ASD) encompass a wide array of debilitating symptoms, including sensory dysfunction and delayed language development. Auditory temporal processing is crucial for speech perception and language development. Abnormal development of temporal processing may account for the language impairments associated with ASD. Very little is known about the development of temporal processing in any animal model of ASD. METHODS: In the current study, we quantify auditory temporal processing throughout development in the Fmr1 knock-out (KO) mouse model of Fragile X Syndrome (FXS), a leading genetic cause of intellectual disability and ASD-associated behaviors. Using epidural electrodes in awake and freely moving wildtype (WT) and KO mice, we recorded auditory event related potentials (ERP) and auditory temporal processing with a gap-in-noise auditory steady state response (gap-ASSR) paradigm. Mice were recorded at three different ages in a cross sectional design: postnatal (p)21, p30 and p60. Recordings were obtained from both auditory and frontal cortices. The gap-ASSR requires underlying neural generators to synchronize responses to gaps of different widths embedded in noise, providing an objective measure of temporal processing across genotypes and age groups. RESULTS: We present evidence that the frontal, but not auditory, cortex shows significant temporal processing deficits at p21 and p30, with poor ability to phase lock to rapid gaps in noise. Temporal processing was similar in both genotypes in adult mice. ERP amplitudes were larger in Fmr1 KO mice in both auditory and frontal cortex, consistent with ERP data in humans with FXS. CONCLUSIONS: These data indicate cortical region-specific delays in temporal processing development in Fmr1 KO mice. Developmental delays in the ability of frontal cortex to follow rapid changes in sounds may shape language delays in FXS, and more broadly in ASD.

Alcohol and lactation: Developmental deficits in a mouse model.

It is well documented that prenatal ethanol exposure via maternal consumption of alcohol during pregnancy alters brain and behavioral development in offspring. Thus, the Centers for Disease Control (CDC) advises against maternal alcohol consumption during pregnancy. However, little emphasis has been placed on educating new parents about alcohol consumption while breastfeeding. This is partly due to a paucity of research on lactational ethanol exposure (LEE) effects in children; although, it has been shown that infants exposed to ethanol via breast milk frequently present with reduced body mass, low verbal IQ scores, and altered sleeping patterns. As approximately 36% of breastfeeding mothers in the US consume alcohol, continued research in this area is critical. Our study employed a novel murine LEE model, where offspring were exposed to ethanol via nursing from postnatal day (P) 6 through P20, a period correlated with infancy in humans. Compared to controls, LEE mice had reduced body weights and neocortical lengths at P20 and P30. Brain weights were also reduced in both ages in males, and at P20 for females, however, female brain weights recovered to control levels by P30. We investigated neocortical features and found that frontal cortex thickness was reduced in LEE males compared to controls. Analyses of dendritic spines in the prelimbic subdivision of medial prefrontal cortex revealed a trend of reduced densities in LEE mice. Results of behavioral tests suggest that LEE mice engage in higher risk-taking behavior, show abnormal stress regulation, and exhibit increased hyperactivity. In summary, our data describe potential adverse brain and behavioral developmental outcomes due to LEE. Thus, women should be advised to refrain from consuming alcohol during breastfeeding until additional research can better guide recommendations of safe maternal practices in early infancy.

Transgenerational Effects of Prenatal Ethanol Exposure in Prepubescent Mice.

Background: Fetal alcohol spectrum disorders (FASD) represent a leading cause of non-genetic neuropathologies. Recent preclinical evidence from suggests that prenatal ethanol exposure (PrEE), like other environmental exposures, may have a significant, transgenerational impact on the offspring of directly exposed animals, including altered neocortical development at birth and behavior in peri-pubescent mice. How these adverse behavioral outcomes are manifested within the brain at the time of behavioral disruption remains unknown. Methods: A transgenerational mouse model of FASD was used to generate up to a third filial generation of offspring to study. Using a multi-modal battery of behavioral assays, we assessed motor coordination/function, sensorimotor processing, risk-taking behavior, and depressive-like behavior in postnatal day (P) 20 pre-pubescent mice. Additionally, sensory neocortical area connectivity using dye tracing, neocortical gene expression using in situ RNA hybridization, and spine density of spiny stellate cells in the somatosensory cortex using Golgi-Cox staining were examined in mice at P20. Results: We found that PrEE induces behavioral abnormalities including abnormal sensorimotor processing, increased risk-taking behavior, and increased depressive-like behaviors that extend to the F3 generation in 20-day old mice. Assessment of both somatosensory and visual cortical connectivity, as well as cortical RZRß expression in pre-pubescent mice yielded no significant differences among any experimental generations. In contrast, only directly-exposed F1 mice displayed altered cortical expression of Id2 and decreased spine density among layer IV spiny stellate cells in somatosensory cortex at this pre-pubescent, post weaning age. Conclusion: Our results suggest that robust, clinically-relevant behavioral abnormalities are passed transgenerationally to the offspring of mice directly exposed to prenatal ethanol. Additionally, in contrast to our previous findings in the newborn PrEE mouse, a lack of transgenerational findings within the brain at this later age illuminates the critical need for future studies to attempt to discover the link between neurological function and the described behavioral changes. Overall, our study suggests that multi-generational effects of PrEE may have a substantial impact on human behavior as well as health and well-being and that these effects likely extend beyond early childhood.

Analysis of Ignition Sites for the Explosives 3,3'-Diamino-4,4'-azoxyfurazan (DAAF) and 1,3,5,7-Tetranitro-1,3,5,7-tetrazoctane (HMX) Using Crush Gun Impact Testing.

The handling safety characteristics of energetic materials must be measured in order to ensure the safe transport and use of explosives. Drop-weight impact sensitivity measurements are one of the first standardized tests performed for energetics. They utilize a small amount of the explosive sample and a standard weight, which is dropped on the material from various heights to determine its sensitivity. While multiple laboratories have used the impact sensitivity test as an initial screening tool for explosive sensitivity for the past 60 years, variability exists due to the use of different instruments, different methods to determine the initiation, and the scatter commonly associated with less-sensitive explosives. For example, standard explosives such as 1,3,5,7-tetranitro-1,3,5,7-tetrazoctane (HMX) initiate reliably and consistently on the drop-weight impact test, whereas insensitive explosives such as 3,3'-diamino-4,4'-azoxyfurazan (DAAF) exhibit variability in sound levels and the expended material. Herein we investigate the impact sensitivity of DAAF and HMX along with a more detailed investigation of ignition sites using a novel "crush gun" apparatus: a pneumatically powered drop-weight tower with advanced diagnostics, including high-speed visual and infrared cameras. Using this crush gun assembly, the ignition sites in HMX and DAAF were analyzed with respect to the effects of particle size and the presence of a source of grit. The formation of ignition sites was observed in both explosives; however, only HMX showed ignition sites that propagated to a deflagration at lower firing speeds. Finally, the presence of grit particles was shown to increase the occurrence of ignition sites in DAAF at lower firing speeds, though propagation to a full reaction was not observed on the time scale of the test. These results enable a better understanding of how ignition and propagation occurs during the impact testing of DAAF.

Probative value of absolute and relative judgments in eyewitness identification.

It is well-accepted that eyewitness identification decisions based on relative judgments are less accurate than identification decisions based on absolute judgments. However, the theoretical foundation for this view has not been established. In this study relative and absolute judgments were compared through simulations of the WITNESS model (Clark, Appl Cogn Psychol 17:629-654, 2003) to address the question: Do suspect identifications based on absolute judgments have higher probative value than suspect identifications based on relative judgments? Simulations of the WITNESS model showed a consistent advantage for absolute judgments over relative judgments for suspect-matched lineups. However, simulations of same-foils lineups showed a complex interaction based on the accuracy of memory and the similarity relationships among lineup members.

Detection of PhIP in grilled chicken entrées at popular chain restaurants throughout California.

Heterocyclic amines (HCAs), compounds formed when meat is cooked at high temperatures particularly through pan frying, grilling, or barbequing, pose a potential carcinogenic risk to the public. It is unclear whether there is any level at which consumption of HCAs can be considered safe. Efforts to measure these compounds mainly include cooking studies under laboratory conditions and some measurement of home-cooked foods, but analysis of commercially cooked foods has been minimal. Attempts to estimate exposure of the public to these compounds must take into consideration dining outside the home, which could result in significant exposure for some individuals. We surveyed at least 9 locations each of 7 popular chain restaurants (McDonald's, Burger King, Chick-fil-A, Chili's, TGI Friday's, Outback Steakhouse, and Applebee's) in California, collecting one or two entrees from each location. Entrees were analyzed for 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) using high-performance liquid chromatography tandem mass spectrometry. All 100 samples contained PhIP. Concentrations were variable within and between entrees and ranged from 0.08 to 43.2 ng/g. When factoring in the weight of the entrees, absolute levels of PhIP reached over 1,000 ng for some entrees. Potential strategies for reducing exposure include the avoidance of meats cooked using methods that are known to form PhIP.

Rule-based extrapolation: a continuing challenge for exemplar models.

Erickson and Kruschke (1998, 2002) demonstrated that in rule-plus-exception categorization, people generalize category knowledge by extrapolating in a rule-like fashion, even when they are presented with a novel stimulus that is most similar to a known exception. Although exemplar models have been found to be deficient in explaining rule-based extrapolation, Rodrigues and Murre (2007) offered a variation of an exemplar model that was better able to account for such performance. Here, we present the results of a new rule-plus-exception experiment that yields rule-like extrapolation similar to that of previous experiments, and yet the data are not accounted for by Rodrigues and Murre's augmented exemplar model. Further, a hybrid rule-and-exemplar model is shown to better describe the data. Thus, we maintain that rule-plus-exception categorization continues to be a challenge for exemplar-only models.

Executive attention and task switching in category learning: evidence for stimulus-dependent representation.

One class of multiple-system models of category learning posits that within a single category-learning task people can learn to utilize different systems with different category representations to classify different stimuli. This is referred to as stimulus-dependent representation (SDR). The use of SDR implies that learners switch from subtask to subtask as trials demand. Thus, the use of SDR can be assessed via slowed response times, following a representation switch. Additionally, the use of SDR requires control of executive attention to keep inactive representations from interfering with the current response. Subjects were given a category learning task composed of one- and two-dimensional substructures. Control of executive attention was measured using a working memory capacity (WMC) task. Subjects most likely to be using SDR showed greater slowing of responses following a substructure switch and a greater correlation between learning performance and WMC. These results provide support for the principle of SDR in category learning and the reliance of SDR on executive attention.

Perceptual match effects in direct tests of memory: the role of contextual fan.

The aim of the present study was to determine whether physical attributes of a memory representation would affect explicit memory performance and, if so, what type of factors would affect the size of a perceptual match effect. Subjects studied words in different, uncommon fonts and were later asked whether the word had been studied earlier. Words could be re-presented in the original font, a font studied with another word, or a font not seen earlier. In two additional experiments, we varied the numbers of words studied in the same unusual font (1 vs. 12 words per font). Recognition memory for the words was better if the test and study fonts matched, and this effect was larger for fonts not shared with other words. Moreover, old judgments were most likely to be classified as remember responses when words were re-presented in the same font when it had not been studied with other words. Although we found a significant effect of levels of processing, this factor did not interact with whether the font matched between study and test. These results are consistent with the predictions of the source of activation confusion model of memory and suggest that perceptual information operates according to the same memory principles as conceptual information.

Rule-based extrapolation in perceptual categorization.

Erickson and Kruschke (1998) provided a demonstration that in certain situations people will classify novel stimuli according to an extrapolated rule, even when the most similar training exemplar is an exception to the rule. This result challenged exemplar models. Nosofsky and Johansen (2000) have called this finding into question by offering an exemplar-based explanation for those data based on the perceptual features of the stimuli. Here, we describe the results of a new experiment that yields results similar to those found previously without the questionable perceptual features: Participants who learn to classify all the training stimuli have patterns of generalization that indicate a combination of rule and exemplar representation. ATRIUM, a hybrid rule and exemplar model (Erickson & Kruschke, 1998), is shown to account for these data much better than ALCOVE, an exemplar model (Kruschke, 1992). Moreover, four alternate exemplar explanations, including one suggested by Nosofsky and Johansen, cannot account for our new findings.

A reexamination of stimulus-frequency effects in recognition: two mirrors for low- and high-frequency pseudowords.

The word-frequency mirror effect (more hits and fewer false alarms for low-frequency than for high-frequency words) has intrigued memory researchers, and multiple accounts have been offered to explain the result. In this study, participants were differentially familiarized to various pseudowords in a familiarization phase that spanned multiple weeks. Recognition tests given during the first week of familiarization replicated a result of W. T. Maddox and W. K. Estes (1997) that failed to show the classic word-frequency mirror effect for pseudowords; however, recognition tests given toward the end of training showed the classic mirror pattern. In addition, a stimulus-frequency mirror effect for "remember" vs. "know" judgments was obtained. These data are consistent with an account of the mirror effect that posits the involvement of dual processes for episodic recognition.