Hyperglycemia and cognitive impairments anticipate the onset of an overt type 2 diabetes-like phenotype in TALLYHO/JngJ mice.

Type 2 Diabetes mellitus (T2DM) is a metabolic disorder characterized by chronic hyperglycemia, resulting from deficits in insulin secretion, insulin action, or both. Whilst the role of insulin in the peripheral nervous system has been ascertained in countless studies, its role in the central nervous system (CNS) is emerging only recently. Brain insulin has been lately associated with brain disorders like Alzheimer's disease, obsessive compulsive disorder, and attention deficit hyperactivity disorder. Thus, understanding the role of insulin as a common risk factor for mental and somatic comorbidities may disclose novel preventative and therapeutic approaches. We evaluated general metabolism (glucose tolerance, insulin sensitivity, energy expenditure, lipid metabolism, and polydipsia) and cognitive capabilities (attention, cognitive flexibility, and memory), in adolescent, young adult, and adult male and female TALLYHO/JngJ mice (TH, previously reported to constitute a valid experimental model of T2DM due to impaired insulin signaling). Adult TH mice have also been studied for alterations in gut microbiota diversity and composition. While TH mice exhibited profound deficits in cognitive flexibility and altered glucose metabolism, we observed that these alterations emerged either much earlier (males) or independent of (females) a comprehensive constellation of symptoms, isomorphic to an overt T2DM-like phenotype (insulin resistance, polydipsia, higher energy expenditure, and altered lipid metabolism). We also observed significant sex-dependent alterations in gut microbiota alpha diversity and taxonomy in adult TH mice. Deficits in insulin signaling may represent a common risk factor for both T2DM and CNS-related deficits, which may stem from (partly) independent mechanisms.

Dynamic evolutionary interplay between ontogenetic skull patterning and whole-head integration.

The arrangement and morphology of the vertebrate skull reflect functional and ecological demands, making it a highly adaptable structure. However, the fundamental developmental and macroevolutionary mechanisms leading to different vertebrate skull phenotypes remain unclear. Here we exploit the morphological diversity of squamate reptiles to assess the developmental and evolutionary patterns of skull variation and covariation in the whole head. Our geometric morphometric analysis of a complex squamate ontogenetic dataset (209 specimens, 169 embryos, 44 species), covering stages from craniofacial primordia to fully ossified bones, reveals that morphological differences between snake and lizard skulls arose gradually through changes in spatial relationships (heterotopy) followed by alterations in developmental timing or rate (heterochrony). Along with dynamic spatiotemporal changes in the integration pattern of skull bone shape and topology with surrounding brain tissues and sensory organs, we identify a relatively higher phenotypic integration of the developing snake head compared with lizards. The eye, nasal cavity and Jacobson's organ are pivotal in skull morphogenesis, highlighting the importance of sensory rearrangements in snake evolution. Furthermore, our findings demonstrate the importance of early embryonic, ontogenetic and tissue interactions in shaping craniofacial evolution and ecological diversification in squamates, with implications for the nature of cranio-cerebral relations across vertebrates.

A systematic review of preclinical studies exploring the role of insulin signalling in executive function and memory.

Beside its involvement in somatic dysfunctions, altered insulin signalling constitutes a risk factor for the development of mental disorders like Alzheimer's disease and obsessive-compulsive disorder. While insulin-related somatic and mental disorders are often comorbid, the fundamental mechanisms underlying this association are still elusive. Studies conducted in rodent models appear well suited to help decipher these mechanisms. Specifically, these models are apt to prospective studies in which causative mechanisms can be manipulated via multiple tools (e.g., genetically engineered models and environmental interventions), and experimentally dissociated to control for potential confounding factors. Here, we provide a narrative synthesis of preclinical studies investigating the association between hyperglycaemia - as a proxy of insulin-related metabolic dysfunctions - and impairments in working and spatial memory, and attention. Ultimately, this review will advance our knowledge on the role of glucose metabolism in the comorbidity between somatic and mental illnesses.

Automation at the service of the study of executive functions in preclinical models.

Cognitive flexibility involves the capability to switch between different perspectives and implement novel strategies upon changed circumstances. The Wisconsin Card Sorting Test (in humans) and the Attentional Set-Shifting Task (ASST, in rodents) evaluate individual capability to acquire a reward-associated rule and subsequently disregard it in favour of a new one. Both tasks entail consecutive stages wherein subjects discriminate between: two stimuli of a given category (simple discrimination, SD); the stimuli of SD confounded by an irrelevant stimulus of a different category (compound discrimination, CD); different stimuli belonging to the SD category (intradimensional shift, IDS); and two stimuli of the confounding category (extradimensional shift, EDS). The ASST is labour intensive, not sufficiently standardised, and prone to experimental error. Here, we tested the validity of a new, commercially available, automated version of ASST (OPERON) in two independent experiments conducted in: different mouse strains (C57BL/6 and CD1 mice) to confirm their differential cognitive capabilities (Experiment 1); and an experimental model of chronic stress (administration of corticosterone in the drinking water; Experiment 2). In both experiments, OPERON confirmed the findings obtained through the manual version. Just as in Experiment 1 both versions captured the deficit of C57BL/6 mice on the reversal of the CD (CDR), so also in Experiment 2 they provided analogous evidence that corticosterone treated mice have a remarkable impairment in the IDS. Thus, OPERON capitalises upon automated phenotyping to overcome the limitation of the manual version of the ASST while providing comparable results.

Reconstructing the origin and early evolution of the snake brain.

Snakes represent one-eighth of terrestrial vertebrate diversity, encompassing various lifestyles, ecologies, and morphologies. However, the ecological origins and early evolution of snakes are controversial topics in biology. To address the paucity of well-preserved fossils and the caveats of osteological traits for reconstructing snake evolution, we applied a different ecomorphological hypothesis based on high-definition brain reconstructions of extant Squamata. Our predictive models revealed a burrowing lifestyle with opportunistic behavior at the origin of crown snakes, reflecting a complex ancestral mosaic brain pattern. These findings emphasize the importance of quantitatively tracking the phenotypic diversification of soft tissues-including the accurate definition of intact brain morphological traits such as the cerebellum-in understanding snake evolution and vertebrate paleobiology. Furthermore, our study highlights the power of combining extant and extinct species, soft tissue reconstructions, and osteological traits in tracing the deep evolution of not only snakes but also other groups where fossil data are scarce.

Insulin and disorders of behavioural flexibility.

Behavioural inflexibility is a symptom of neuropsychiatric and neurodegenerative disorders such as Obsessive-Compulsive Disorder, Autism Spectrum Disorder and Alzheimer's Disease, encompassing the maintenance of a behaviour even when no longer appropriate. Recent evidence suggests that insulin signalling has roles apart from its regulation of peripheral metabolism and mediates behaviourally-relevant central nervous system (CNS) functions including behavioural flexibility. Indeed, insulin resistance is reported to generate anxious, perseverative phenotypes in animal models, with the Type 2 diabetes medication metformin proving to be beneficial for disorders including Alzheimer's Disease. Structural and functional neuroimaging studies of Type 2 diabetes patients have highlighted aberrant connectivity in regions governing salience detection, attention, inhibition and memory. As currently available therapeutic strategies feature high rates of resistance, there is an urgent need to better understand the complex aetiology of behaviour and develop improved therapeutics. In this review, we explore the circuitry underlying behavioural flexibility, changes in Type 2 diabetes, the role of insulin in CNS outcomes and mechanisms of insulin involvement across disorders of behavioural inflexibility.

Long-term consequences of reduced availability and compensatory supplementation of sialylated HMOs on cognitive capabilities.

Breast milk (BM) is the optimal source of nutrition for mammals' early life. It exerts multiple benefits, including the development of cognitive capabilities and protection against several diseases like obesity and infection of the respiratory tract. However, which components of BM are involved in individual development has remained elusive. Sialylated human milk oligosaccharides (HMOs) may constitute a valid candidate, whereby they represent the principal source of sialic acid and act as building blocks for brain development. We hypothesize that the reduced availability of two HMOs, sialyl(alpha2,6)lactose (6'SL) and sialyl(alpha2,3)lactose (3'SL), may impair attention, cognitive flexibility, and memory in a preclinical model and that the exogenous supplementation of these compounds may contrast the observed deficits. We evaluated cognitive capabilities in a preclinical model exposed to maternal milk containing reduced concentrations of 6'SL and 3'SL during lactation. To modulate their concentrations, we utilized a preclinical model characterized by the absence of genes that synthesize 3'SL and 6'SL (B6.129-St3gal4 tm1.1Jxm and St6gal1tm2Jxm , double genetic deletion), producing milk lacking 3'SL and 6'SL. Then, to ensure exposure to 3'SL-6'SL-poor milk in early life, we adopted a cross-fostering protocol. The outcomes assessed in adulthood were different types of memory, attention and information processing, some of which are part of executive functions. Then, in the second study, we evaluated the long-term compensatory potential of the exogenous oral supplementation of 3'SL and 6'SL during lactation. In the first study, exposure to HMO-poor milk resulted in reduced memory and attention. Specifically, it resulted in impaired working memory in the T-maze test, in reduced spatial memory in the Barnes maze, and in impaired attentional capabilities in the Attentional set-shifting task. In the second part of the study, we did not observe any difference between experimental groups. We hypothesize that the experimental procedures utilized for the exogenous supplementation may have impacted our ability to observe the cognitive read-out in vivo. This study suggests that early life dietary sialylated HMOs play a crucial role in the development of cognitive functions. Future studies are needed to clarify if an exogenous supplementation of these oligosaccharides may compensate for these affected phenotypes.

Endocast, brain, and bones: Correspondences and spatial relationships in squamates.

Vertebrate endocasts are widely used in the fields of paleoneurology and comparative neuroanatomy. The validity of endocranial studies is dependent upon the extent to which an endocast reflects brain morphology. Due to the variable neuroanatomical resolution of vertebrate endocasts, direct information about the brain morphology can be sometimes difficult to assess and needs to be investigated across lineages. Here, we employ X-ray computed tomography (CT), including diffusible iodine-based contrast-enhanced CT, to qualitatively compare brains and endocasts in different species of squamates. The relative position of the squamate brain within the skull, as well as the variability that may exist in such spatial relationships, was examined to help clarify the neurological regions evidence on their endocasts. Our results indicate that squamate endocasts provide variable representation of the brain, depending on species and neuroanatomical regions. The olfactory bulbs and peduncles, cerebral hemispheres, as well as the medulla oblongata represent the most easily discernable brain regions from squamate endocasts. In contrast, the position of the optic lobes, the ventral diencephalon and the pituitary may be difficult to determine depending on species. Finally, squamate endocasts provide very limited or no information about the cerebellum. The spatial relationships revealed here between the brain and the surrounding bones may help to identify each of the endocranial region. However, as one-to-one correspondences between a bone and a specific region appear limited, the exact delimitation of these regions may remain challenging according to species. This study provides a basis for further examination and interpretation of squamate endocast disparity.

A landmarking protocol for geometric morphometric analysis of squamate endocasts.

Landmark-based geometric morphometrics is widely used to study the morphology of the endocast, or internal mold of the braincase, and the diversity associated with this structure across vertebrates. Landmarks, as the basic unit of such methods, are intended to be points of correspondence, selected depending on the question at hand, whose proper definition is essential to guarantee robustness and reproducibility of results. In this study, 20 landmarks are defined to provide a framework to analyze the morphological variability in squamate endocasts. Ten species representing a cross-section of the diversity of Squamata from both phylogenetic and ecological (i.e., habitat) perspectives were considered, to select landmarks replicable throughout the entire clade, regardless of the degree of neuroanatomical resolution of the endocast. To assess the precision, accuracy, and repeatability of these newly defined landmarks, both intraobserver and interobserver error were investigated. Estimates of measurement error show that most of the landmarks established here are highly replicable, and preliminary results suggest that they capture aspects of endocast shape related to both phylogenetic and ecologic signals. This study provides a basis for further examinations of squamate endocast disparity using landmark-based geometric morphometrics.

Game theoretical trajectory planning enhances social acceptability of robots by humans.

Since humans and robots are increasingly sharing portions of their operational spaces, experimental evidence is needed to ascertain the safety and social acceptability of robots in human-populated environments. Although several studies have aimed at devising strategies for robot trajectory planning to perform safe motion in populated environments, a few efforts have measured to what extent a robot trajectory is accepted by humans. Here, we present a navigation system for autonomous robots that ensures safety and social acceptability of robotic trajectories. We overcome the typical reactive nature of state-of-the-art trajectory planners by leveraging non-cooperative game theory to design a planner that encapsulates human-like features of preservation of a personal space, recognition of groups, sequential and strategized decision making, and smooth obstacle avoidance. Social acceptability is measured through a variation of the Turing test administered in the form of a survey questionnaire to a pool of 691 participants. Comparison terms for our tests are a state-of-the-art navigation algorithm (Enhanced Vector Field Histogram, VFH) and purely human trajectories. While all participants easily recognized the non-human nature of VFH-generated trajectories, the distinction between game-theoretical trajectories and human ones were hardly revealed. Our results mark a strong milestone toward the full integration of robots in social environments.

Developmental differences in the impact of perceptual salience on short-term memory performance and meta-memory skills.

In everyday life, individuals are surrounded by many stimuli that compete to access attention and memory. Evidence shows that perceptually salient stimuli have more chances to capture attention resources, thus to be encoded into short-term memory (STM). However, the impact of perceptual salience on STM at different developmental stages is entirely unexplored. Here we assessed STM performance and meta-memory skills of 6, 10, and 18 years-old participants (total N = 169) using a delayed match-to-sample task. On each trial, participants freely explored a complex (cartoon-like) scene for 4 s. After a retention interval of 4 s, they discriminated the same/different position of a target-object extracted from the area of maximal or minimal salience of the initially-explored scene. Then, they provided a confidence judgment of their STM performance, as an index of meta-memory skills. When taking into account 'confident' responses, we found increased STM performance following targets at maximal versus minimal salience only in adult participants. Similarly, only adults showed enhanced meta-memory capabilities following maximal versus minimal salience targets. These findings documented a late development in the impact of perceptual salience on STM performance and in the improvement of metacognitive capabilities to properly judge the content of one's own memory representation.

Altered Hippocampal Resting-state Functional Connectivity in Highly Superior Autobiographical Memory.

Individuals with Highly Superior Autobiographical Memory (HSAM) provide the opportunity to investigate the neurobiological substrates of enhanced memory performance. While previous studies started to assess the neural correlates of memory retrieval in HSAM, here we assessed for the first time the intrinsic connectivity of a core memory region, the hippocampus, with the whole brain, in 8 HSAM subjects (HSAMs) and 21 controls during resting-state functional neuroimaging. We found in HSAMs vs. controls disrupted hippocampal resting-state functional connectivity (rsFC) with high-level control regions belonging to the saliency network (the anterior cingulate cortex and the left and right insulae), and to the ventral fronto-parietal attentional network (the temporo-parietal junction and the inferior frontal gyrus), also involved with salience detection. Conversely, HSAMs showed enhanced hippocampal rsFC with sensory regions along the fusiform gyrus and the inferior temporal cortex. This altered pattern of hippocampal rsFC might be interpreted as a reduced capability of HSAMs to discriminate and select salient information, with a subsequent increase in the probability to encode and consolidate sensory information irrespective of their task-relevancy. Ultimately, these findings provide evidence that HSAM might be paradoxically enabled by an altered hippocampal rsFC that bypasses regions involved with salience detection in favor of specialized sensory regions.

Searching for a Relationship between Early Breastfeeding and Cognitive Development of Attention and Working Memory Capacity.

Previous research consistently reported that subjects that were exclusively breastfed (eBF) vs. not-exclusively breastfed (neBF) during infancy (0-6 months) showed increased scores of general intelligence measures (e.g., the intelligence quotient). However, the existent literature largely neglected whether breastfeeding also affects specific cognitive processes, such as attention and working memory (WM) capacity. We tested whether eBF vs. neBF subjects showed performance differences in relation to these two core cognitive functions. The Attention Network Test (ANT), to measure alerting, orienting, and conflict, and the Change Colour Task (CCT), to measure visuospatial WM capacity, were administered to 144 participants divided according to age (6-, 10-, and 18-year-old participants) and breastfeeding (eBF or neBF during 0-6 months of life). Importantly, the sub-groups were homogenous in terms of maternal education, a factor potentially affecting the relation between breastfeeding and cognition. While we found increased performance as a function of participants' age in both tasks, we failed to observe effects related to breastfeeding, as evidenced by Bayesian analyses. These findings highlight for the first time that the pattern of nutrition provided during early infancy does not appear to affect the development of attention and WM capacity, at least starting from the age considered in the present study.

Exposure to 3'Sialyllactose-Poor Milk during Lactation Impairs Cognitive Capabilities in Adulthood.

Breast milk exerts pivotal regulatory functions early in development whereby it contributes to the maturation of brain and associated cognitive functions. However, the specific components of maternal milk mediating this process have remained elusive. Sialylated human milk oligosaccharides (HMOs) represent likely candidates since they constitute the principal neonatal dietary source of sialic acid, which is crucial for brain development and neuronal patterning. We hypothesize that the selective neonatal lactational deprivation of a specific sialylated HMOs, sialyl(alpha2,3)lactose (3'SL), may impair cognitive capabilities (attention, cognitive flexibility, and memory) in adulthood in a preclinical model. To operationalize this hypothesis, we cross-fostered wild-type (WT) mouse pups to B6.129-St3gal4tm1.1Jxm/J dams, knock-out (KO) for the gene synthesizing 3'SL, thereby providing milk with approximately 80% 3'SL content reduction. We thus exposed lactating WT pups to a selective reduction of 3'SL and investigated multiple cognitive domains (including memory and attention) in adulthood. Furthermore, to account for the underlying electrophysiological correlates, we investigated hippocampal long-term potentiation (LTP). Neonatal access to 3'SL-poor milk resulted in decreased attention, spatial and working memory, and altered LTP compared to the control group. These results support the hypothesis that early-life dietary sialylated HMOs exert a long-lasting role in the development of cognitive functions.

Collective Emotional Contagion in Zebrafish.

Seeking to match our emotional state with one of those around us is known as emotional contagion-a fundamental biological process that underlies social behavior across several species and taxa. While emotional contagion has been traditionally considered to be a prerogative of mammals and birds, recent findings are demonstrating otherwise. Here, we investigate emotional contagion in groups of zebrafish, a freshwater model species which is gaining momentum in preclinical studies. Zebrafish have high genetic homology to humans, and they exhibit a complex behavioral repertoire amenable to study social behavior. To investigate whether individual emotional states can be transmitted to group members, we pharmacologically modulated anxiety-related behaviors of a single fish through Citalopram administration and we assessed whether the altered emotional state spread to a group of four untreated conspecifics. By capitalizing upon our in-house developed tracking algorithm, we successfully preserved the identity of all the subjects and thoroughly described their individual and social behavioral phenotypes. In accordance with our predictions, we observed that Citalopram administration consistently reduced behavioral anxiety of the treated individual, in the form of reduced geotaxis, and that such a behavioral pattern readily generalized to the untreated subjects. A transfer entropy analysis of causal interactions within the group revealed that emotional contagion was directional, whereby the treated individual influenced untreated subjects, but not vice-versa. This study offers additional evidence that emotional contagion is biologically preserved in simpler living organisms amenable to preclinical investigations.

Highly superior autobiographical memory in aging: A single case study.

Whilst countless studies have shown that aging is associated with cognitive decline in the general population, near to nothing is known about this association in elderly individuals naturally exhibiting enhanced memory capabilities. The identification of a 75 years old individual (GC) with highly superior autobiographical memory (HSAM), and his willingness to volunteer to our study over a period of five years, allowed us to investigate this issue in a single case study. At the age of 75 years, GC was screened for HSAM with the Public Events Quiz and the Random Dates Quiz, with a positive outcome. GC's memory performance was extraordinarily higher than normal-memory control subjects (>3 standard deviations), and comparable to a group of younger HSAM individuals (mean age of 32.5 years; Santangelo et al., 2018). GC underwent general neuropsychological (Mini-Mental State Examination), personality (Personality Assessment Inventory), and brain morphological (brain volumes and lesions) assessments, showing no deviation from normal ranges. To gain insight into the brain mechanisms underlying his memory performance, GC underwent functional brain imaging during the retrieval of memories associated with random dates. The latter were also rated in terms of reliving quality and emotional valence. Similar to younger HSAM individuals, GC's access to past memories recruited a wide network of prefrontal and temporo-parietal regions, especially during the recollection of memories associated with a lower reliving rating, suggesting a compensatory mechanism in HSAM. Increased activity in the insula was instead associated with emotionally-positive memories. Five years later, GC was tested again for HSAM and showed no sign of memory decline, whereby his memory performance was indistinguishable from the tests he performed five years earlier. GC's case suggests that highly superior memory performance can manifest without apparent decline in physiological aging. Implications of the current findings for the extant models of autobiographical memory are discussed.

Sialylated human milk oligosaccharides program cognitive development through a non-genomic transmission mode.

Breastmilk contains bioactive molecules essential for brain and cognitive development. While sialylated human milk oligosaccharides (HMOs) have been implicated in phenotypic programming, their selective role and underlying mechanisms remained elusive. Here, we investigated the long-term consequences of a selective lactational deprivation of a specific sialylated HMO in mice. We capitalized on a knock-out (KO) mouse model (B6.129-St6gal1tm2Jxm/J) lacking the gene responsible for the synthesis of sialyl(alpha2,6)lactose (6'SL), one of the two sources of sialic acid (Neu5Ac) to the lactating offspring. Neu5Ac is involved in the formation of brain structures sustaining cognition. To deprive lactating offspring of 6'SL, we cross-fostered newborn wild-type (WT) pups to KO dams, which provide 6'SL-deficient milk. To test whether lactational 6'SL deprivation affects cognitive capabilities in adulthood, we assessed attention, perseveration, and memory. To detail the associated endophenotypes, we investigated hippocampal electrophysiology, plasma metabolomics, and gut microbiota composition. To investigate the underlying molecular mechanisms, we assessed gene expression (at eye-opening and in adulthood) in two brain regions mediating executive functions and memory (hippocampus and prefrontal cortex, PFC). Compared to control mice, WT offspring deprived of 6'SL during lactation exhibited consistent alterations in all cognitive functions addressed, hippocampal electrophysiology, and in pathways regulating the serotonergic system (identified through gut microbiota and plasma metabolomics). These were associated with a site- (PFC) and time-specific (eye-opening) reduced expression of genes involved in central nervous system development. Our data suggest that 6'SL in maternal milk adjusts cognitive development through a short-term upregulation of genes modulating neuronal patterning in the PFC.

Acute Citalopram administration alters zebrafish social dynamics in a behavioral teleporting experiment.

In the Star Trek universe, Scotty can "beam you up" and teleport you to a location. Even though science fiction is yet to be reality, is teleporting still possible? Recently, we have developed a robotic platform that is capable of teleporting the behavior of fish from one tank to another through biologically-inspired robotic replicas which map the behavior of a fish from a tank to the other one. Our results indicate that behavioral teleporting is a promising tool to study the biological determinants of behavior. Here we demonstrate its use in pharmacological research, by investigating the effects of selective serotonin reuptake inhibitors of social interactions.

Acute Citalopram administration modulates anxiety in response to the context associated with a robotic stimulus in zebrafish.

BACKGROUND: Anxiety represents one of the most urgent health challenges in Western Countries, where it is associated with major medical and societal costs. A common therapeutic approach is the use of selective serotonin reuptake inhibitors, such as Citalopram. However, this treatment of choice is characterized by incomplete efficacy and potential side effects. Preclinical research is needed to detail the mechanisms underlying therapeutic efficacy of available treatments. METHODS: Zebrafish, a rapidly emerging model species, constitutes an excellent candidate for high-throughput studies in behavioral pharmacology. Here, we present a robotics-based experimental paradigm to investigate the effects of acute Citalopram administration on conditioned place aversion. We trained adult subjects in a three-partitioned tank, consisting of one central and two lateral compartments: the latter were associated either with a fear eliciting robotic stimulus or with an empty environment. Following training, we implemented an automated three-dimensional tracking system to assess the spatial association and detail individual phenotype in a stimulus-free test session. RESULTS: We observed a linear dose-response profile with respect to geotaxis, with increasing Citalopram concentrations reducing the tendency to swim near the bottom of the tank. Although control subjects failed to exhibit the predicted conditioned aversion, we found preliminary evidence that Citalopram may affect sexes differentially, with male subjects showing increased conditioned aversion at low Citalopram concentration. CONCLUSIONS: Experimental paradigms based on robotics and three-dimensional tracking can contribute methodological advancements in zebrafish behavioral psychopharmacology.