EASI-FISH for thick tissue defines lateral hypothalamus spatio-molecular organization.

Determining the spatial organization and morphological characteristics of molecularly defined cell types is a major bottleneck for characterizing the architecture underpinning brain function. We developed Expansion-Assisted Iterative Fluorescence In Situ Hybridization (EASI-FISH) to survey gene expression in brain tissue, as well as a turnkey computational pipeline to rapidly process large EASI-FISH image datasets. EASI-FISH was optimized for thick brain sections (300 µm) to facilitate reconstruction of spatio-molecular domains that generalize across brains. Using the EASI-FISH pipeline, we investigated the spatial distribution of dozens of molecularly defined cell types in the lateral hypothalamic area (LHA), a brain region with poorly defined anatomical organization. Mapping cell types in the LHA revealed nine spatially and molecularly defined subregions. EASI-FISH also facilitates iterative reanalysis of scRNA-seq datasets to determine marker-genes that further dissociated spatial and morphological heterogeneity. The EASI-FISH pipeline democratizes mapping molecularly defined cell types, enabling discoveries about brain organization.

The Allen Mouse Brain Common Coordinate Framework: A 3D Reference Atlas.

Recent large-scale collaborations are generating major surveys of cell types and connections in the mouse brain, collecting large amounts of data across modalities, spatial scales, and brain areas. Successful integration of these data requires a standard 3D reference atlas. Here, we present the Allen Mouse Brain Common Coordinate Framework (CCFv3) as such a resource. We constructed an average template brain at 10 µm voxel resolution by interpolating high resolution in-plane serial two-photon tomography images with 100 µm z-sampling from 1,675 young adult C57BL/6J mice. Then, using multimodal reference data, we parcellated the entire brain directly in 3D, labeling every voxel with a brain structure spanning 43 isocortical areas and their layers, 329 subcortical gray matter structures, 81 fiber tracts, and 8 ventricular structures. CCFv3 can be used to analyze, visualize, and integrate multimodal and multiscale datasets in 3D and is openly accessible (https://atlas.brain-map.org/).

Artificial intelligence's interpretation of the neuroanatomical aspect of Peter Paul Rubens's copy of "The Battle of Anghiari" by Leonardo da Vinci.

We tested to see how Ruben's copy of "The Battle of Anghiari" by Leonardo da Vinci would be interpreted by AI in a neuroanatomical aspect. We used WOMBO Dream, an artificial intelligence (AI)-based algorithm that creates images based on words and figures. The keyword we provided for the algorithm was "brain" and the reference image was Ruben's drawing. AI interpreted the whole drawing as a representation of the brain. The image generated by the algorithm was similar to our interpretation of the same painting.

Interindividual differences in intergenerational sustainable behavior are associated with cortical thickness of the dorsomedial and dorsolateral prefrontal cortex.

Intergenerational sustainability requires people of the present generation to make sacrifices today to benefit others of future generations (e.g. mitigating climate change, reducing public debt). Individuals vary greatly in their intergenerational sustainability, and the cognitive and neural sources of these interindividual differences are not yet well understood. We here combined neuroscientific and behavioral methods by assessing interindividual differences in cortical thickness and by using a common-pool resource paradigm with intergenerational contingencies. This enabled us to look for objective, stable, and trait-like neural markers of interindividual differences in consequential intergenerational behavior. We found that individuals behaving sustainably (vs. unsustainably) were marked by greater cortical thickness of the dorsomedial and dorsolateral prefrontal cortex. Given that these brain areas are involved in perspective-taking and self-control and supported by mediation analyses, we speculate that greater cortical thickness of these brain areas better enable individuals to take the perspective of future generations and to resist temptations to maximize personal benefits that incur costs for future generations. By meeting recent calls for the contribution of neuroscience to sustainability research, it is our hope that the present study advances the transdisciplinary understanding of interindividual differences in intergenerational sustainability.

Who initiates punishment, who joins punishment? Disentangling types of third-party punishers by neural traits.

The act of punishing unfair behavior by unaffected observers (i.e., third-party punishment) is a crucial factor in the functioning of human societies. In everyday life, we see different types of individuals who punish. While some individuals initiate costly punishment against an unfair person independently of what other observers do (independent punishers), others condition their punishment engagement on the presence of another person who punishes (conditional punishers). Still others do not want to partake in any sort of punishment (nonpunishers). Although these distinct behavioral types have a divergent impact on human society, the sources of heterogeneity are poorly understood. We present novel laboratory evidence on the existence of these three types. We use anatomical brain characteristics in combination with stated motives to characterize these types. Findings revealed that independent punishers have larger gray matter volume in the right temporo-parietal junction compared to conditional punishers and nonpunishers, an area involved in social cognition. Conditional punishers are characterized by larger gray matter volume in the right dorsolateral prefrontal cortex, a brain area known to be involved in behavioral control and strategic reasoning, compared to independent punishers and nonpunishers. Finally, both independent punishers and nonpunishers are characterized by larger gray matter volume in an area involved in the processing of social and monetary rewards, that is, the bilateral caudate. By using a neural trait approach, we were able to differentiate these three types clearly based on their neural signatures, allowing us to shed light on the underlying psychological mechanisms.

Examining volumetric gradients based on the frustum surface ratio in the brain in autism spectrum disorder.

Autism spectrum disorder (ASD) is a complex neurodevelopmental disorder that is accompanied by neurodevelopmental differences in regional cortical volume (CV), and a potential layer-specific pathology. Conventional measures of CV, however, do not indicate how volume is distributed across cortical layers. In a sample of 92 typically developing (TD) controls and 92 adult individuals with ASD (aged 18-52 years), we examined volumetric gradients by quantifying the degree to which CV is weighted from the pial to the white surface of the brain. Overall, the spatial distribution of Frustum Surface Ratio (FSR) followed the gyral and sulcal pattern of the cortex and approximated a bimodal Gaussian distribution caused by a linear mixture of vertices on gyri and sulci. Measures of FSR were highly correlated with vertex-wise estimates of mean curvature, sulcal depth, and pial surface area, although none of these features explained more than 76% variability in FSR on their own. Moreover, in ASD, we observed a pattern of predominant increases in the degree of FSR relative to TD controls, with an atypical neurodevelopmental trajectory. Our findings suggest a more outward-weighted gradient of CV in ASD, which may indicate a larger contribution of supragranular layers to regional differences in CV.

Altered Gesture Imitation and Brain Anatomy in Adult Prader-Willi Syndrome Patients.

OBJECTIVE: To explore motor praxis in adults with Prader-Willi syndrome (PWS) in comparison with a control group of people with intellectual disability (ID) and to examine the relationship with brain structural measurements. METHOD: Thirty adult participants with PWS and 132 with ID of nongenetic etiology (matched by age, sex, and ID level) were assessed using a comprehensive evaluation of the praxis function, which included pantomime of tool use, imitation of meaningful and meaningless gestures, motor sequencing, and constructional praxis. RESULTS: Results support specific praxis difficulties in PWS, with worse performance in the imitation of motor actions and better performance in constructional praxis than ID peers. Compared with both control groups, PWS showed increased gray matter volume in sensorimotor and subcortical regions. However, we found no obvious association between these alterations and praxis performance. Instead, praxis scores correlated with regional volume measures in distributed apparently normal brain areas. CONCLUSIONS: Our findings are consistent in showing significant impairment in gesture imitation abilities in PWS and, otherwise, further indicate that the visuospatial praxis domain is relatively preserved. Praxis disability in PWS was not associated with a specific, focal alteration of brain anatomy. Altered imitation gestures could, therefore, be a consequence of widespread brain dysfunction. However, the specific contribution of key brain structures (e.g., areas containing mirror neurons) should be more finely tested in future research.

Patterns of Cortical Folding Associated with Autistic Symptoms in Carriers and Noncarriers of the 22q11.2 Microdeletion.

22q11.2 deletion syndrome (22q11.2DS) is a genetic condition accompanied by a range of psychiatric manifestations, including autism spectrum disorder (ASD). It remains unknown, however, whether these symptoms are mediated by the same or distinct neural mechanisms as in idiopathic ASD. Here, we examined differences in lGI associated with ASD in 50 individuals with 22q11.2DS (n = 25 with ASD, n = 25 without ASD) and 81 individuals without 22q11.2DS (n = 40 with ASD, n = 41 typically developing controls). We initially utilized a factorial design to identify the set of brain regions where lGI is associated with the main effect of 22q11.2DS, ASD, and with the 22q11.2DS-by-ASD interaction term. Subsequently, we employed canonical correlation analysis (CCA) to compare the multivariate association between variability in lGI and the complex clinical phenotype of ASD between 22q11.2DS carriers and noncarriers. Across approaches, we established that even though there is a high degree of clinical similarity across groups, the associated patterns of lGI significantly differed between carriers and noncarriers of the 22q11.2 microdeletion. Our results suggest that ASD symptomatology recruits different neuroanatomical underpinnings across disorders and that 22q11.2DS individuals with ASD represent a neuroanatomically distinct subgroup that differs from 22q11.2DS individuals without ASD and from individuals with idiopathic ASD.

Mapping the superficial morphology of the occipital lobe: proposal of a universal nomenclature for clinical and anatomical use.

The superficial anatomy of the occipital lobe has been described as irregular and highly complex. This notion mainly arises from the variability of the regional sulco-gyral architecture. Our aim was to investigate the prevalence, morphology, and correlative anatomy of the sulci and gyri of the occipital region in cadaveric specimens and to summarize the nomenclature used in the literature to describe these structures. To this end, 33 normal, adult, formalin-fixed hemispheres were studied. In addition, a review of the relevant literature was conducted with the aim to compare our findings with data from previous studies. Hence, in the lateral occipital surface, we recorded the lateral occipital sulcus and the intraoccipital sulcus in 100%, the anterior occipital sulcus in 24%, and the inferior occipital sulcus in 15% of cases. In the area of the occipital pole, we found the transverse occipital sulcus in 88% of cases, the lunate sulcus in 64%, the occipitopolar sulcus in 24%, and the retrocalcarine sulcus in 12% of specimens. In the medial occipital surface, the calcarine fissure and parieto-occipital sulcus were always present. Finally, the basal occipital surface was always indented by the posterior occipitotemporal and posterior collateral sulci. A sulcus not previously described in the literature was identified on the supero-lateral aspect of the occipital surface in 85% of cases. We named this sulcus "marginal occipital sulcus" after its specific topography. In this study, we offer a clear description of the occipital surface anatomy and further propose a standardized taxonomy for clinical and anatomical use.

The Neuroanatomy of Autism Spectrum Disorder Symptomatology in 22q11.2 Deletion Syndrome.

22q11.2 Deletion Syndrome (22q11.2DS) is a genetic condition associated with a high prevalence of neuropsychiatric conditions that include autism spectrum disorder (ASD). While evidence suggests that clinical phenotypes represent distinct neurodevelopmental outcomes, it remains unknown whether this translates to the level of neurobiology. To fractionate the 22q11.2DS phenotype on the level of neuroanatomy, we examined differences in vertex-wise estimates of cortical volume, surface area, and cortical thickness between 1) individuals with 22q11.2DS (n = 62) and neurotypical controls (n = 57) and 2) 22q11.2DS individuals with ASD symptomatology (n = 30) and those without (n = 25). We firstly observed significant differences in surface anatomy between 22q11.2DS individuals and controls for all 3 neuroanatomical features, predominantly in parietotemporal regions, cingulate and dorsolateral prefrontal cortices. We also established that 22q11.2DS individuals with ASD symptomatology were neuroanatomically distinct from 22q11.2DS individuals without ASD symptoms, particularly in brain regions that have previously been linked to ASD (e.g., dorsolateral prefrontal cortices and the entorhinal cortex). Our findings indicate that different clinical 22q11.2DS phenotypes, including those with ASD symptomatology, may represent different neurobiological subgroups. The spatially distributed patterns of neuroanatomical differences associated with ASD symptomatology in 22q11.2DS may thus provide useful information for patient stratification and the prediction of clinical outcomes.

Design and construction of an innovative brain phantom prototype for MRI.

PURPOSE: The purpose of this project was to construct a physical brain phantom for MRI, mimicking structure and T1 relaxation properties of white matter (WM) and gray matter (GM). METHODS: The phantom design comprised 2 compartments, 1 resembling the WM and 1 resembling the GM. Their T1 relaxation times, as assessed using an inversion recovery turbo spin echo sequence, were reproduced using an agar gel doped with contrast agent (CA) and their folding patterns were simulated through a molding-casting procedure using 3D-printed casts and flexible silicone molds. Three versions of the assembling procedure were adopted to build: Phantom1 without any separation; Phantom2 with a varnish layer; and Phantom3 with a thin wax layer between the compartments. RESULTS: Phantom1 was characterized by an immediate diffusion of CA between the 2 compartments. Phantom2 and Phantom3, instead, showed relaxation times and shape comparable with the target ones identified in a healthy control subject (WM: 754 ± 40 ms; GM: 1277 ± 96 ms). Moreover, both compartments revealed intact gyri and sulci. However, the diffusion of CA made Phantom2 stable only for a short period of time. Phantom3 showed stability within a time window of several days but the wax layer between the WM and GM was visible in the MRI. CONCLUSION: Structural and intensity properties of the constructed phantoms are useful in evaluating and validating steps from image acquisition to image processing. Moreover, the described constructing procedure and its modular design make it adjustable to a variety of applications.

Variation in sexual brain size dimorphism over the breeding cycle in the three-spined stickleback.

Snapshot analyses have demonstrated dramatic intraspecific variation in the degree of brain sexual size dimorphism (SSD). Although brain SSD is believed to be generated by the sex-specific cognitive demands of reproduction, the relative roles of developmental and population-specific contributions to variation in brain SSD remain little studied. Using a common garden experiment, we tested for sex-specific changes in brain anatomy over the breeding cycle in three-spined stickleback (Gasterosteus aculeatus) sampled from four locations in northern Europe. We found that the male brain increased in size (ca. 24%) significantly more than the female brain towards breeding, and that the resulting brain SSD was similar (ca. 20%) for all populations over the breeding cycle. Our findings support the notion that the stickleback brain is highly plastic and changes over the breeding cycle, especially in males, likely as an adaptive response to the cognitive demands of reproduction (e.g. nest construction and parental care). The results also provide evidence to suggest that breeding-related changes in brain size may be the reason for the widely varying estimates of brain SSD across studies of this species, cautioning against interpreting brain size measurements from a single time point as fixed/static.

Neuronal morphology alterations in autism and possible role of oxytocin.

Current understanding of the neuroanatomical abnormalities in autism includes gross anatomical changes in several brain areas and microstructural alterations in neuronal cells as well. There are many controversies in the interpretation of the imaging data, evaluation of volume and size of particular brain areas, and their functional translation into a broad autism phenotype. Critical questions of neuronal pathology in autism include the concept of the reversible plasticity of morphological changes, volume alterations of brain areas, and both short- and long-term consequences of adverse events present during the brain development. At the cellular level, remodeling of the actin cytoskeleton is considered as one of the critical factors associated with the autism spectrum disorders. Alterations in the composition of the neuronal cytoskeleton, in particular abnormalities in the polymerization of actin filaments and their associated proteins underlie the functional consequences in behavior resulting in symptoms and clinical correlates of autism spectrum disorder. In the present review, a special attention is devoted to the role of oxytocin in experimental models of neurodevelopmental disorders manifesting alterations in neuronal morphology.

Down syndrome is accompanied by significantly reduced cortical grey-white matter tissue contrast.

Increased cortical thickness (CT) has been reported in Down syndrome (DS) during childhood and adolescence, but it remains unclear, which components of the neural architecture underpin these increases and if CT remains altered in adults. Among other factors, differences in CT measures could be driven by reduced tissue contrast between grey and white matter (GWC), which has been reported in neurodegenerative disorders, such as Alzheimer's disease. Using structural magnetic resonance imaging, we therefore examined differences in CT and GWC in 26 adults with DS, and 23 controls, to (1) examine between-group differences in CT in adulthood, (2) establish whether DS is associated with significant reductions in GWC, and (3) determine the influence of GWC variability on between-group differences in CT. As hypothesized, we observed that DS was accompanied by wide-spread increases in CT, and significantly reduced GWC in several large clusters distributed across the cortex. Out of all vertices with a significant between-group difference in CT, 38.50% also displayed a significant reduction in GWC. This percentage of overlap was also statistically significant and extremely unlikely to be obtained by chance (p = .0002). Differences in GWC thus seem to explain some, although not all, of the differences in CT observed in DS. In addition, our study is the first to extend previous in vivo reports of altered CT in DS during childhood and adolescence to older adults, implying that the regional pattern of neuroanatomical differences associated with DS remains stable across the lifespan.

Brain anatomy of the 4-day-old European rabbit.

The European rabbit (Oryctolagus cuniculus) is a widely used model in fundamental, medical and veterinary neurosciences. Besides investigations in adults, rabbit pups are relevant to study perinatal neurodevelopment and early behaviour. To date, the rabbit is also the only species in which a pheromone - the mammary pheromone (MP) - emitted by lactating females and active on neonatal adaptation has been described. The MP is crucial since it contributes directly to nipple localisation and oral seizing in neonates, i.e. to their sucking success. It may also be one of the non-photic cues arising from the mother, which stimulates synchronisation of the circadian system during pre-visual developmental stages. Finally, the MP promotes neonatal odour associative and appetitive conditioning in a remarkably rapid and efficient way. For these different reasons, the rabbit offers a currently unique opportunity to determine pheromonal-induced brain processing supporting adaptation early in life. Therefore, it is of interest to create a reference work of the newborn rabbit pup brain, which may constitute a tool for future multi-disciplinary and multi-approach research in this model, and allow comparisons related to the neuroethological basis of social and feeding behaviour among newborns of various species. Here, in line with existing experimental studies, and based on original observations, we propose a functional anatomical description of brain sections in 4-day-old rabbits with a particular focus on seven brain regions which appear important for neonatal perception of sensory signals emitted by the mother, circadian adaptation to the short and single daily nursing of the mother in the nest, and expression of specific motor actions involved in nipple localisation and milk intake. These brain regions involve olfactory circuits, limbic-related areas important in reward, motivation, learning and memory formation, homeostatic areas engaged in food anticipation, and regions implicated in circadian rhythm and arousal, as well as in motricity.

Neuroanatomical distribution and function of the vasopressin V1B receptor in the rat brain deciphered using specific fluorescent ligands.

It is now accepted that vasopressin, through V1A/V1B receptors, centrally regulates cognitive functions such as memory, affiliation, stress, fear and depression. However, the respective roles of these receptor isoforms and their contribution to stress-related pathologies remain uncertain. The development of new therapeutic treatments requires a precise knowledge of the distribution of these receptors within the brain, which has been so far hampered by the lack of selective V1B markers. In the present study, we have determined the pharmacological properties of three new potent rat V1B fluorescent ligands and demonstrated that they constitute valuable tools for simultaneous visualization and activation of native V1B receptors in living rat brain tissue. Thus, d[Leu4,Lys-Alexa 647)8]VP (analogue 3), the compound with the best affinity-selectivity/fluorescence ratio for the V1B receptor emerged as the most promising. The rat brain regions most concerned by stress such as hippocampus, olfactory bulbs, cortex and amygdala display the highest V1B fluorescent labelling with analogue 3. In the hippocampus CA2, V1B receptors are located on glutamatergic, not GABAergic neurones, and are absent from astrocytes. Using AVP-EGFP rats, we demonstrate the presence of V1B autoreceptors on AVP-secreting neurones not only in the hypothalamus, but also sparsely in the hippocampus. Finally, using both electrophysiology and visualization of ERK phosphorylation, we show analogue 3-induced activation of the V1B receptor in situ. This will help to analyse expression and functionality of V1B receptors in the brain and contribute to further explore the AVPergic circuitry in normal and pathological conditions.

New species of the genus Spectracanthicus (Loricariidae, Hypostominae, Ancistrini) from the Rio Javaés (Rio Araguaia basin), with a description of gross brain morphology.

A new species of Spectracanthicus is described from the Rio Javaés, Rio Araguaia basin. The new species is distinguished from its congeners (except Spectracanthicus immaculatus) by colour pattern: body dark grey to dark brown without dots or blotches (v. body colour with yellowish small dots in Spectracanthicus murinus, Spectracanthicus punctatissimus and Spectracanthicus tocantinensis and large white dots in Spectracanthicus zuanoni). It can be further distinguished from S. immaculatus by having thicker and less numerous teeth, with up to eight premaxillary and 20 dentary teeth (v. teeth thinner and more numerous with up to 22 premaxillary and 30 dentary teeth); dorsal and caudal fins without curved spines (v. dorsal and caudal fins with curved spines). Other osteological characters can also diagnose the new species from its congeners. In addition, a gross brain description and brief comments on the new species' ecological habitat are given.

Inter-individual variations and hemispheric asymmetries in structural connectivity patterns of the inferior fronto-occipital fascicle: a diffusion tensor imaging tractography study.

PURPOSE: Precise knowledge of the structural connectivity of white matter fascicles could yield new insights into function and is important for neurosurgical planning. Therefore, we aimed to provide a detailed map of the cortical terminations of the inferior fronto-occipital fascicle (IFOF), with special emphasis on putative inter-individual variations and hemispheric asymmetries. METHODS: Deterministic diffusion tensor imaging-based tractography was used to perform virtual dissection of the IFOF in 20 healthy subjects. The IFOF was probed from a single seed region of interest placed within the external/extreme capsule, i.e. the white matter region of "obligatory passage" along the known path of the IFOF. This enabled to reconstruct all the fibers belonging to the IFOF and to provide the complete map of their cortical terminations. RESULTS: We observed widespread projections over a total of 11 cortical territories within the occipital, parietal, temporal and frontal lobes. Importantly, compared to previous studies we consistently found some inter-individual variability with several distinct patterns connecting subsets of the 11 cortical territories, and tangible differences between the two hemispheres. IFOF terminations within the superior parietal lobule were rightward lateralized, whereas terminations within the inferior frontal gyrus were leftward lateralized. CONCLUSIONS: Our results provide a clinically relevant map of IFOF's cortical terminations, including intra- and inter-individual variations. Right-left differences in connectivity patterns might be related to known functional asymmetries in the human brain, and reinforce the general evidence that the IFOF likely supports distinct clinical features and functional roles according to the (affected) hemisphere, such as language and spatial attention.

Individual Differences in Cognitive Control Circuit Anatomy Link Sensation Seeking, Impulsivity, and Substance Use.

Individuals vary widely in their tendency to seek stimulation and act impulsively, early developing traits with genetic origins. Failures to regulate these behaviors increase risk for maladaptive outcomes including substance abuse. Here, we explored the neuroanatomical correlates of sensation seeking and impulsivity in healthy young adults. Our analyses revealed links between sensation seeking and reduced cortical thickness that were preferentially localized to regions implicated in cognitive control, including anterior cingulate and middle frontal gyrus (n = 1015). These associations generalized to self-reported motor impulsivity, replicated in an independent group (n = 219), and correlated with heightened alcohol, tobacco, and caffeine use. Critically, the relations between sensation seeking and brain structure were evident in participants without a history of alcohol or tobacco use, suggesting that observed associations with anatomy are not solely a consequence of substance use. These results demonstrate that individual differences in the tendency to seek stimulation, act on impulse, and engage in substance use are correlated with the anatomical structure of cognitive control circuitry. Our findings suggest that, in healthy populations, covariation across these complex multidimensional behaviors may in part originate from a common underlying biology. SIGNIFICANCE STATEMENT: Impaired cognitive control may result in a tendency to seek stimulation impulsively and an increased risk for maladaptive outcomes, including substance abuse. Here, we examined the structural correlates of sensation seeking and impulsivity in a large cohort of healthy young adults. Our analyses revealed links between sensation seeking and reduced cortical thickness that were preferentially localized to regions implicated in cognitive control, including anterior cingulate and middle frontal gyrus. The observed associations generalized to motor impulsivity, replicated in an independent group, and predicted heightened alcohol, tobacco, and caffeine use. These data indicate that normal variability in cognitive control system anatomy predicts sensation seeking and motor impulsivity in the healthy populations, potentially increasing risk for substance use disorders.

Reconstructing anatomy from electro-physiological data.

Here we show how it is possible to make estimates of brain structure based on MEG data. We do this by reconstructing functional estimates onto distorted cortical manifolds parameterised in terms of their spherical harmonics. We demonstrate that both empirical and simulated MEG data give rise to consistent and plausible anatomical estimates. Importantly, the estimation of structure from MEG data can be quantified in terms of millimetres from the true brain structure. We show, for simulated data, that the functional assumptions which are closer to the functional ground-truth give rise to anatomical estimates that are closer to the true anatomy.