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The postnatal development of ocular dominance columns (ODCs) in monkey visual cortex provides an exquisite model for studying mechanisms of experience-guided neuronal plasticity. While the presence of columns at birth in Old World monkeys is now well established, it remains unclear whether cortical neurons at this early stage are capable of modulating gene expression in response to changing sensory conditions. Using a set of monocular deprivation and stimulation protocols, we examined activity-driven expression of the immediate-early genes (IEGs) c-fos and zif268 during the critical period, starting as early as the first postnatal day. The expression levels are similar in layers II/III, IVC and VI throughout development, with no selective decline in the thalamorecepient layer (layer IVC) of adult monkeys. A narrow strip of non-columnar c-Fos expression was observed at the border of layers IVC and V. Our results show that neurons in monkey visual cortex are equipped at birth with the molecular machinery for coupling sensory inputs to active genomic responses and that this responsivity extends throughout the critical period. The findings are discussed within the context of a possible role for IEGs in sensory-driven cortical plasticity during development. [ABSTRACT FROM AUTHOR]

Urban areas in North American cities with positive trends in bicycle usage also witness a high number of cyclist injuries every year. Previous cyclist safety studies based on the traditional approach, which relies on historical crash data, are known to have some limitations such as the fact that crashes need to happen (a reactive approach). This paper explores the use of GPS deceleration events as a surrogate-proactive measure and investigates the relationship between reported cyclist road injuries and deceleration events. The surrogate safety measure is defined based on deceleration values representing hard breaking situations. This work uses a large sample of GPS cyclist trip data from a smartphone application to extract deceleration rates at intersections and along segments and to explore its relationship with the number of observed injuries and validate deceleration rate (DR) as a surrogate safety measure. Using Spearman's rank correlation coefficient, we compared the ranking of sites based on the expected number of injuries and based on DR. The ranks of expected injuries and dangerous decelerations were found to have a correlation of 0.60 at signalized intersections, 0.53 at non-signalized intersections and 0.57 at segments. Despite the promising results of this study, more granular data and validation work needs to be done to improve the reliability of the measures. The technological limitations and future work are discussed at the end of the paper., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

In the literature, a crash-based modeling approach has long been used to evaluate the factors that contribute to cyclist injury risk at intersections. However, this approach has been criticized as crashes are required to occur before contributing factors can be identified and countermeasures can be implemented. Moreover, human factors related to dangerous behaviors are difficult to evaluate using crash-based methods. As an alternative, surrogate safety measures have been developed to address the issue of reliance on crash data. Despite recent developments, few methodologies and little empirical evidence exist on bicycle-vehicle interactions at intersections using video-based data and statistical analyses to identify associated factors. This study investigates bicycle-vehicle conflict severity and evaluates the impact of different factors, including gender, on cyclist risk at urban intersections with cycle tracks. A segmented ordered logit model is used to evaluate post-encroachment time between cyclists and vehicles. Video data was collected at seven intersections in Montreal, Canada. Road user trajectories were automatically extracted, classified, and filtered using a computer vision software to yield 1514 interactions. The discrete choice variable was generated by dividing post-encroachment time into normal interactions, conflicts, and dangerous conflicts. Independent variables reflecting attributes of the cyclist, vehicle, and environment were extracted either automatically or manually. Results indicated that an ordered model is appropriate for analyzing traffic conflicts and identifying key factors. Furthermore, exogenous segmentation was beneficial in comparing different segments of the population within a single model. Male cyclists, with all else being equal, were less likely than female cyclists to be involved in conflicts and dangerous conflicts at the studied intersections. Bicycle and vehicle speed, along with the time of the conflict relative to the red light phase, were other significant factors in conflict severity. These results will contribute to and further the understanding of gender differences in cycling within North America., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Cities in North America have been building bicycle infrastructure, in particular cycle tracks, with the intention of promoting urban cycling and improving cyclist safety. These facilities have been built and expanded but very little research has been done to investigate the safety impacts of cycle tracks, in particular at intersections, where cyclists interact with turning motor-vehicles. Some safety research has looked at injury data and most have reached the conclusion that cycle tracks have positive effects of cyclist safety. The objective of this work is to investigate the safety effects of cycle tracks at signalized intersections using a case-control study. For this purpose, a video-based method is proposed for analyzing the post-encroachment time as a surrogate measure of the severity of the interactions between cyclists and turning vehicles travelling in the same direction. Using the city of Montreal as the case study, a sample of intersections with and without cycle tracks on the right and left sides of the road were carefully selected accounting for intersection geometry and traffic volumes. More than 90h of video were collected from 23 intersections and processed to obtain cyclist and motor-vehicle trajectories and interactions. After cyclist and motor-vehicle interactions were defined, ordered logit models with random effects were developed to evaluate the safety effects of cycle tracks at intersections. Based on the extracted data from the recorded videos, it was found that intersection approaches with cycle tracks on the right are safer than intersection approaches with no cycle track. However, intersections with cycle tracks on the left compared to no cycle tracks seem to be significantly safer. Results also identify that the likelihood of a cyclist being involved in a dangerous interaction increases with increasing turning vehicle flow and decreases as the size of the cyclist group arriving at the intersection increases. The results highlight the important role of cycle tracks and the factors that increase or decrease cyclist safety. Results need however to be confirmed using longer periods of video data., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Children with fetal alcohol spectrum disorders display behavioural and intellectual impairments that strongly implicate dysfunction within the frontal cortex. Deficits in social behaviour and cognition are amongst the most pervasive outcomes of prenatal ethanol exposure. Our naturalistic vervet monkey model of fetal alcohol exposure (FAE) provides an unparalleled opportunity to study the neurobehavioral outcomes of prenatal ethanol exposure in a controlled experimental setting. Recent work has revealed a significant reduction of the neuronal population in the frontal lobes of these monkeys. We used an intersensory matching procedure to investigate audiovisual perception of socially relevant stimuli in young FAE vervet monkeys. Here we show a domain-specific deficit in audiovisual integration of socially relevant stimuli. When FAE monkeys were shown a pair of side-by-side videos of a monkey concurrently presenting two different calls along with a single audio track matching the content of one of the calls, they were not able to match the correct video to the single audio track. This was manifest by their average looking time being equally spent towards both the matching and non-matching videos. However, a group of normally developing monkeys exhibited a significant preference for the non-matching video. This inability to integrate and thereby discriminate audiovisual stimuli was confined to the integration of faces and voices as revealed by the monkeys' ability to match a dynamic face to a complex tone or a black-and-white checkerboard to a pure tone, presumably based on duration and/or onset-offset synchrony. Together, these results suggest that prenatal ethanol exposure negatively affects a specific domain of audiovisual integration. This deficit is confined to the integration of information that is presented by the face and the voice and does not affect more elementary aspects of sensory integration.

Recent studies suggest that exposure to only one component of audiovisual events can lead to cross-modal cortical activation. However, it is not certain whether such crossmodal recruitment can occur in the absence of explicit conditioning, semantic factors, or long-term associations. A recent study demonstrated that crossmodal cortical recruitment can occur even after a brief exposure to bimodal stimuli without semantic association. In addition, the authors showed that the primary visual cortex is under such crossmodal influence. In the present study, we used molecular activity mapping of the immediate early gene zif268. We found that animals, which had previously been exposed to a combination of auditory and visual stimuli, showed increased number of active neurons in the primary visual cortex when presented with sounds alone. As previously implied, this crossmodal activation appears to be the result of implicit associations of the two stimuli, likely driven by their spatiotemporal characteristics; it was observed after a relatively short period of exposure (~45 min) and lasted for a relatively long period after the initial exposure (~1 day). These results suggest that the previously reported findings may be directly rooted in the increased activity of the neurons occupying the primary visual cortex.

Hemiparesis, unilateral weakness or partial paralysis, is a common outcome following hemispherectomy in humans. We use the non-human primate as an invaluable translational model for our understanding of developmental plasticity in response to hemispherectomy. Three infant vervet monkeys (Chlorocebus sabeus) underwent hemispherectomy at a median age of 9 weeks and two additional monkeys at 48 months. Gross motor assessment was conducted in a large open field that contained a horizontal bar spanning the width of the cage. Subjects were assessed yearly following surgery in infantile lesions for a period of 3 years. Adult-lesioned subjects were assessed 40 months following surgery. Shortly after surgery both infant and adult-lesioned subjects were unable to move the contralateral side of their body, but all subjects were able to walk within 6 months following surgery. At each time point the lower limb gait was normal in infant-lesioned subjects with no apparent limp or dragging, however the upper limb demonstrated significant impairment. Horizontal bar crossing was significantly impaired during the first 24 months following surgery. Adult-lesioned subjects also displayed upper limb movement impairments similar to infant-lesioned subjects. In addition the adult-lesioned subjects displayed a noticeable lower limb limp, which was not observed in the infant-lesioned group. Both groups at each time point showed a propensity for ipsiversive turning. The upper limb gait impairment and horizontal bar crossing of lesioned subjects are reminiscent of hemiparesis seen in hemisperectomized humans with the young-lesioned subjects showing a greater propensity for recovery., ((c) 2009 Elsevier Ireland Ltd. All rights reserved.)

Several lines of evidence suggest that exposure to only one component of typically audiovisual events can lead to crossmodal cortical activation. These effects are likely explained by long-term associations formed between the auditory and visual components of such events. It is not certain whether such crossmodal recruitment can occur in the absence of explicit conditioning, semantic factors, or long-term association; nor is it clear whether primary sensory cortices can be recruited in such paradigms. In the present study we tested the hypothesis that crossmodal cortical recruitment would occur even after a brief exposure to bimodal stimuli without semantic association. We used positron emission tomography, and an apparatus allowing presentation of spatially and temporally congruous audiovisual stimuli (noise bursts and light flashes). When presented with only the auditory or visual components of the bimodal stimuli, naïve subjects showed only modality-specific cortical activation, as expected. However, subjects who had previously been exposed to the audiovisual stimuli showed increased cerebral blood flow in the primary visual cortex when presented with sounds alone. Functional connectivity analysis suggested that the auditory cortex was the source of visual cortex activity. This crossmodal activation appears to be the result of implicit associations of the two stimuli, likely driven by their spatiotemporal characteristics; it was observed after a relatively short period of exposure (approximately 45 min), and lasted for a relatively long period after the initial exposure (approximately 1 day). The findings indicate that auditory and visual cortices interact with one another to a larger degree than typically assumed., (2009 Elsevier Ltd. All rights reserved.)

Immunohistochemistry (IHC) is one of the most widely used laboratory techniques for the detection of target proteins in situ. Questions concerning the expression pattern of a target protein across the entire brain are relatively easy to answer when using IHC in small brains, such as those of rodents. However, answering the same questions in large and convoluted brains, such as those of primates presents a number of challenges. Here we present a systematic approach for immunodetection of target proteins in an adult monkey brain. This approach relies on the tissue embedding and sectioning methodology of NeuroScience Associates (NSA) as well as tools developed specifically for batch-staining of free-floating sections. It results in uniform staining of a set of sections which, at a particular interval, represents the entire brain. The resulting stained sections can be subjected to a wide variety of analytical procedures in order to measure protein levels, the population of neurons expressing a certain protein.

Unbiased stereology is a method for accurately and efficiently estimating the total neuron number (or other cell type) in a given area of interest(1). To achieve this goal 6-10 systematic sections should be probed covering the entire structure. Typically this involves processing 1/5 sections which leaves a significant amount of material unprocessed. In order to maximize the material, we propose an inexpensive method for preserving fixed tissue as part of a long-term storage research plan. As tissue is sliced and processed for the desired stain or antibody, alternate sections should be systematically placed in antigen preserve at -20 degrees C for future processing. Using 24-well plates, sections can be placed in order for future retrieval. Using this method, tissue can be stored and processed for immunohistochemistry over the course of years.

The use of non-human primates provides an excellent translational model for our understanding of developmental and aging processes in humans(1-6). In addition, the use of non-human primates has recently afforded the opportunity to naturally model complex psychiatric disorders such as alcohol abuse(7). Here we describe a technique for blocking the brain in the coronal plane of the vervet monkey (Chlorocebus aethiops sabeus) in the intact skull in stereotaxic space. The method described here provides a standard plane of section between blocks and subjects and minimizes partial sections between blocks. Sectioning a block of tissue in the coronal plane also facilitates the delineation of an area of interest. This method provides manageable sized blocks since a single hemisphere of the vervet monkey yields more than 1200 sections when slicing at 50 microm. Furthermore by blocking the brain into 1cm blocks, it facilitates penetration of sucrose for cyroprotection and allows the block to be sliced on a standard cryostat.

The visual system in humans is considered the gateway to the world and plays a principal role in the plethora of sensory, perceptual and cognitive processes. It is therefore not surprising that quality of vision is tied to quality of life . Despite widespread clinical and basic research surrounding the causes of visual disorders, many forms of visual impairments, such as retinitis pigmentosa and macular degeneration, lack effective treatments. Non-human primates have the closest general features of eye development to that of humans. Not only do they have a similar vascular anatomy, but amongst other mammals, primates have the unique characteristic of having a region in the temporal retina specialized for high visual acuity, the fovea(1). Here we describe a general technique for dissecting the primate retina to provide tissue for retinal histology, immunohistochemistry, laser capture microdissection, as well as light and electron microscopy. With the extended use of the non-human primate as a translational model, our hope is that improved understanding of the retina will provide insights into effective approaches towards attenuating or reversing the negative impact of visual disorders on the quality of life of affected individuals.

The non-human primate is an important translational species for understanding the normal function and disease processes of the human brain. Unbiased stereology, the method accepted as state-of-the-art for quantification of biological objects in tissue sections, generates reliable structural data for biological features in the mammalian brain. The key components of the approach are unbiased (systematic-random) sampling of anatomically defined structures (reference spaces), combined with quantification of cell numbers and size, fiber and capillary lengths, surface areas, regional volumes and spatial distributions of biological objects within the reference space. Among the advantages of these stereological approaches over previous methods is the avoidance of all known sources of systematic (non-random) error arising from faulty assumptions and non-verifiable models. This study documents a biological application of computerized stereology to estimate the total neuronal population in the frontal cortex of the vervet monkey brain (Chlorocebus aethiops sabeus), with assistance from two commercially available stereology programs, BioQuant Life Sciences and Stereologer (Figure 1). In addition to contrast and comparison of results from both the BioQuant and Stereologer systems, this study provides a detailed protocol for the Stereologer system.

Fragile X syndrome (FXS) is one of the most prevalent forms of heritable mental retardation and developmental delay in males. The syndrome is caused by the silencing of a single gene (fragile X mental retardation-1; FMR1) and the lack of expression of its protein product (fragile X mental retardation-1 protein; FMRP). Recent work has linked the high expression levels of FMRP in the magnocellular layers of lateral geniculate nucleus (M-LGN) of the visual system to a specific reduction of perceptual function known to be mediated by that neural structure. This finding has given rise to the intriguing notion that FMRP expression level may be used as an index of susceptibility of specific brain regions to the observed perceptual and cognitive deficits in FXS. We undertook a comprehensive expression profiling study of FMRP in the monkey to obtain further insight into the link between FMPR expression and the behavioural impact of its loss in FXS. We report here the first 3D whole-brain map of FMRP expression in the Old-World monkey and show that certain brain structures display high FMRP levels, such as the cerebellum, striatum, and temporal lobe structures. This finding provides support for the notion that FMRP expression loss is linked to behavioural and cognitive impairment associated with these structures. We argue that whole-brain FMRP expression mapping may be used to formulate and test new hypotheses about other forms of impairments in FXS that were not specifically examined in this study.

Background: Understanding the evolutionary origins of a phenotype requires understanding the relationship between ontogenetic and phylogenetic processes. Human infants have been shown to undergo a process of perceptual narrowing during their first year of life, whereby their intersensory ability to match the faces and voices of another species declines as they get older. We investigated the evolutionary origins of this behavioral phenotype by examining whether or not this developmental process occurs in non-human primates as well., Methodology/principal Findings: We tested the ability of infant vervet monkeys (Cercopithecus aethiops), ranging in age from 23 to 65 weeks, to match the faces and voices of another non-human primate species (the rhesus monkey, Macaca mulatta). Even though the vervets had no prior exposure to rhesus monkey faces and vocalizations, our findings show that infant vervets can, in fact, recognize the correspondence between rhesus monkey faces and voices (but indicate that they do so by looking at the non-matching face for a greater proportion of overall looking time), and can do so well beyond the age of perceptual narrowing in human infants. Our results further suggest that the pattern of matching by vervet monkeys is influenced by the emotional saliency of the Face+Voice combination. That is, although they looked at the non-matching screen for Face+Voice combinations, they switched to looking at the matching screen when the Voice was replaced with a complex tone of equal duration. Furthermore, an analysis of pupillary responses revealed that their pupils showed greater dilation when looking at the matching natural face/voice combination versus the face/tone combination., Conclusions/significance: Because the infant vervets in the current study exhibited cross-species intersensory matching far later in development than do human infants, our findings suggest either that intersensory perceptual narrowing does not occur in Old World monkeys or that it occurs later in development. We argue that these findings reflect the faster rate of neural development in monkeys relative to humans and the resulting differential interaction of this factor with the effects of early experience.

Background: It is believed that a face-specific system exists within the primate ventral visual pathway that is separate from a domain-general nonface object coding system. In addition, it is believed that hemispheric asymmetry, which was long held to be a distinct feature of the human brain, can be found in the brains of other primates as well. We show here for the first time by way of a functional imaging technique that face- and object-selective neurons form spatially distinct clusters at the cellular level in monkey inferotemporal cortex. We have used a novel functional mapping technique that simultaneously generates two separate activity profiles by exploiting the differential time course of zif268 mRNA and protein expression., Results: We show that neurons activated by face stimulation can be visualized at cellular resolution and distinguished from those activated by nonface complex objects. Our dual-activity maps of face and object selectivity show that face-selective patches of various sizes (mean, 22.30 mm2; std, 32.76 mm2) exist throughout the IT cortex in the context of a large expanse of cortical territory that is responsive to visual objects., Conclusions: These results add to recent findings that face-selective patches of various sizes exist throughout area IT and provide the first direct anatomical evidence at cellular resolution for a hemispheric asymmetry in favor of the right hemisphere. Together, our results support the notion that human and monkey brains share a similarity in both anatomical organization and distribution of function with respect to high-level visual processing.

Immediate-Early Genes are a class of genes that are rapidly up-regulated following neural stimulation. Due to their quality as potential activity markers in the CNS, they have been used extensively in functional mapping studies. At least three genes have been popularly used, including zif268 (Egr1, NGFI, Krox-24, or ZENK), c-fos and recently, Arc. A number of techniques have been developed in applying IEG labelling for the development of functional maps, thus overcoming some of the earlier limitations of this approach. Current developments highlight the future prospects of cellular-resolution functional activity mapping of the brain.

The use of inducible transcription factors for mapping neural activity is now a common procedure. We have previously developed a double-labelling technique that allows visualization of activated neurons after two different stimulation sequences. The technique exploits the differential time course of mRNA versus protein expression of transcription factors. However, the precise details of the differential time course remained unknown. Here, we provide a complete up- and downregulation profile for both the c-fos and zif268 genes, as determined through combined in situ hybridization and immunocytochemical detection of the mRNA and protein products in primary visual cortex. The data presented here can be used in the design of future studies employing double-label mapping of neural activation following a compound stimulus.

A monoclonal antibody that recognizes a nonphosphorylated epitope on the medium and high molecular weight subunits of neurofilament (NF) proteins was used to investigate laminar and cell morphology changes in monkey striate cortex during post-natal development. Six cortices were obtained from monkeys of a variety of ages: five from developing animals with ages spanning the critical period and one adult. At post-natal day (PD) 0, immunohistochemistry with the SMI-32 antibody revealed immunoreactive (IR) cells in layer IVB and in infragranular layer VI. Early in the critical period (PD 7), these layers become more defined with an increase in the density of immunopositive cells. At the height of the critical period (PD 30 and 42), a drastic increase in the density of SMI-32 labelled pyramidal neurons in layers V and VI was observed. Similarly, layer IVC showed an abundance of dendritic fragments and dendrites that appeared to originate from the infragranular layers. At the end of the critical period (PD 103), a trend toward morphological maturation for individual neurons found within each layer was observed. During any developmental time point, neurons at first appearance tended to show an immature morphology with staining largely restricted to the cell bodies. As such, the characteristic arborizations common to mature pyramidal and multipolar cells was not evident. We propose that the staining pattern seen in this study is consistent with the idea that layers anatomically associated with the magnocellular (M) pathway develop earlier than their parvocellular (P) counterparts.

The rapid accumulation of inducible transcription factors (ITFs), such as c-Fos and Zif268, in activated neurons combined with histological methods that offer detection at the cellular level are key features that have led to their wide use in visualizing activated neurons. There are two major drawbacks of ITFs that limit their use in the CNS--cell-type expression specificity and stimulus-transcription coupling uncertainty. Recent technical advances in the field of molecular activity mapping now permit dual-labeling approaches that help resolve some of these ambiguities and identify neurons that are activated by different sensory stimuli. Furthermore, the recent identification of the robl/LC7-like gene, which shows immediate-early repression after stimulation, may have utility in functional mapping where it can be used to delineate quiescent neurons and serve as a complement to molecular activity markers.

The AP-1 transcription factor, which is composed of various combinations of Fos and Jun proteins, is believed to be a key participant in molecular processes that guide activity-dependent changes in gene expression. In this study, we investigated the activity of different MAP kinases that have been implicated in AP-1 activation. We examined the activities of ERK, JNK/SAPK, and p38 MAPK along with their nuclear targets (Elk-1 and c-Jun) in rat visual cortex after light stimulation. The transcription factor Elk-1 (a possible regulator of c-fos expression) was found to be transiently modified by phosphorylation when visual stimulation was applied after a period of dark rearing. In vitro kinase assay with Elk-1 as substrate showed that light stimulation activated MAPK/ERK in visual cortex but not frontal cortex. Furthermore, ERK activation was temporally matched to onset of Elk-1 phosphorylation. The activity of JNK1 (c-Jun N-terminal kinase 1) was elevated at 2-6 h after visual exposure and was also temporally correlated to increase of endogenous P-c-Jun levels and its appearance within the AP-1 DNA-binding complex. The activities of p38 MAP kinases did not change significantly. These results demonstrate the differential engagement of MAPK signaling pathways following sensory stimulation and their relative effects upon AP-1 expression in the intact brain., (Copyright 1999 Academic Press.)

The postnatal development of ocular dominance columns (ODCs) in monkey visual cortex provides an exquisite model for studying mechanisms of experience-guided neuronal plasticity. While the presence of columns at birth in Old World monkeys is now well established, it remains unclear whether cortical neurons at this early stage are capable of modulating gene expression in response to changing sensory conditions. Using a set of monocular deprivation and stimulation protocols, we examined activity-driven expression of the immediate-early genes (IEGs) c-fos and zif268 during the critical period of development. We observed well-delineated patterns of ODCs produced by sensory regulation of both IEGs throughout the critical period, starting as early as the first postnatal day. The expression levels are similar in layers II/II, IVC and VI throughout development, with no selective decline in the thalamorecepient layer (layer IVC) of adult monkeys. A narrow strip of non-columnar c-Fos expression was observed at the border of layers IVC and V. Our results show that neurons in monkey visual cortex are equipped at birth with the molecular machinery for coupling sensory inputs to active genomic responses and that this responsivity extends throughout the critical period. The findings are discussed within the context of a possible role for IEGs in sensory-driven cortical plasticity during development.