The importance of understanding the behavioural phenotypes of genetic syndromes associated with intellectual disability

Behavioural phenotype research is of benefit to a large number of children with genetic syndromes and associated developmental delay. This article presents an overview of this research area and demonstrates how understanding pathways between gene disorders and behaviour can inform our understanding of the difficulties individuals with genetic syndromes and developmental delay experience, including self-injurious behaviour, social exploitation, social anxiety, social skills deficits, sensory differences, temper outbursts and repetitive behaviours. In addition, physical health difficulties and their interaction with behaviour are considered. The article demonstrates the complexity involved in assessing a child with a rare genetic syndrome.

Evaluation of methods and applications for behavioural profiling of transgenic mice

During the last 10-15 years interest in mouse behavioural analysis has evolved considerably. The driving force is development in molecular biological techniques that allow manipulation of the mouse genome by changing the expression of genes. Therefore, with some limitations it is possible to study how genes participate in regulation of physiological functions and to create models explaining genetic contribution to various pathological conditions. The first aim of our study was to establish a framework for behavioural phenotyping of genetically modified mice. We established comprehensive battery of tests for the initial screening of mutant mice. These included tests for exploratory and locomotor activity, emotional behaviour, sensory functions, and cognitive performance. Our interest was in the behavioural patterns of common background strains used for genetic manipulations in mice. Additionally we studied the behavioural effect of sex differences, test history, and individual housing. Our findings highlight the importance of careful consideration of genetic background for analysis of mutant mice. It was evident that some backgrounds may mask or modify the behavioural phenotype of mutants and thereby lead to false positive or negative findings. Moreover, there is no universal strain that is equally suitable for all tests, and using different backgrounds allows one to address possible phenotype modifying factors. We discovered that previous experience affected performance in several tasks. The most sensitive traits were the exploratory and emotional behaviour, as well as motor and nociceptive functions. Therefore, it may be essential to repeat some of the tests in naïve animals for assuring the phenotype. Social isolation for a long time period had strong effects on exploratory behaviour, but also on learning and memory. All experiments revealed significant interactions between strain and environmental factors (test history or housing condition) indicating genotype-dependent effects of environmental manipulations. Several mutant line analyses utilize this information. For example, we studied mice overexpressing as well as those lacking extracellular matrix protein heparin-binding growth-associated molecule (HB-GAM), and mice lacking N-syndecan (a receptor for HB-GAM). All mutant mice appeared to be fertile and healthy, without any apparent neurological or sensory defects. The lack of HB-GAM and N-syndecan, however, significantly reduced the learning capacity of the mice. On the other hand, overexpression of HB-GAM resulted in facilitated learning. Moreover, HB-GAM knockout mice displayed higher anxiety-like behaviour, whereas anxiety was reduced in HB-GAM overexpressing mice. Changes in hippocampal plasticity accompanied the behavioural phenotypes. We conclude that HB-GAM and N-syndecan are involved in the modulation of synaptic plasticity in hippocampus and play a role in regulation of anxiety- and learning-related behaviour.

Temper outbursts in Prader-Willi syndrome: causes, behavioural and emotional sequence and responses by carers

Background: Temper outbursts are common in Prader-Willi syndrome but rarely described in detail. This study investigated the phenomenology of temper outbursts in terms of antecedents, sequence of behaviours and emotions and intervention strategies used.

Method: A semi-structured interview about temper outbursts was conducted with the main carers of seven children (9.5 to 16.7 years) and seven adults (24.7 to 47.10 years) with Prader-Willi syndrome (10 male, 4 female). Reliability and validity of the interview results was established.

Results: Various setting events increased and reduced the likelihood of temper outbursts. The most common antecedent was a change to routine or expectation. There were marked similarities in the sequence of behaviours and emotions during temper outbursts, with anger rising quickly followed by expressions of remorse and distress at the end of an outburst.

Discussion: The sequence of behaviours and emotions within outbursts was similar to that described in temper tantrums in typical development. Cognitive and emotional processes are likely to be important in the understanding of temper outbursts with implications for early intervention.

Epigenetic mechanisms mediating vulnerability and resilience to psychiatric disorders

The impact that stressful encounters have upon long-lasting behavioural phenotypes is varied. Whereas a significant proportion of the population will develop "stress-related" conditions such as post-traumatic stress disorder or depression in later life, the majority are considered "resilient" and are able to cope with stress and avoid such psychopathologies. The reason for this heterogeneity is undoubtedly multi-factorial, involving a complex interplay between genetic and environmental factors. Both genes and environment are of critical importance when it comes to developmental processes, and it appears that subtle differences in either of these may be responsible for altering developmental trajectories that confer vulnerability or resilience. At the molecular level, developmental processes are regulated by epigenetic mechanisms, with recent clinical and pre-clinical data obtained by ourselves and others suggesting that epigenetic differences in various regions of the brain are associated with a range of psychiatric disorders, including many that are stress-related. Here we provide an overview of how these epigenetic differences, and hence susceptibility to psychiatric disorders, might arise through exposure to stress-related factors during critical periods of development.

A population-specific HTR2B stop codon predisposes to severe impulsivity

Impulsivity, describing action without foresight, is an important feature of several psychiatric diseases, suicidality and violent behaviour. The complex origins of impulsivity hinder identification of the genes influencing it and the diseases with which it is associated. Here we perform exon-focused sequencing of impulsive individuals in a founder population, targeting fourteen genes belonging to the serotonin and dopamine domain. A stop codon in HTR2B was identified that is common (minor allele frequency > 1%) but exclusive to Finnish people. Expression of the gene in the human brain was assessed, as well as the molecular functionality of the stop codon, which was associated with psychiatric diseases marked by impulsivity in both population and family-based analyses. Knockout of Htr2b increased impulsive behaviours in mice, indicative of predictive validity. Our study shows the potential for identifying and tracing effects of rare alleles in complex behavioural phenotypes using founder populations, and indicates a role for HTR2B in impulsivity.

A specific pathway can be identified between genetic characteristics and behaviour profiles in Prader-Willi syndrome via cognitive, environmental and physiological mechanisms

Behavioural phenotypes associated with genetic syndromes have been extensively investigated in order to generate rich descriptions of phenomenology, determine the degree of specificity of behaviours for a particular syndrome, and examine potential interactions between genetic predispositions for behaviour and environmental influences. However, relationships between different aspects of behavioural phenotypes have been less frequently researched and although recent interest in potential cognitive phenotypes or endophenotypes has increased, these are frequently studied independently of the behavioural phenotypes.

Taking Prader-Willi syndrome (PWS) as an example, we discuss evidence suggesting specific relationships between apparently distinct aspects of the PWS behavioural phenotype and relate these to specific endophenotypic characteristics.

The framework we describe progresses through biological, cognitive, physiological and behavioural levels to develop a pathway from genetic characteristics to behaviour with scope for interaction with the environment at any stage.

We propose this multilevel approach as useful in setting out hypotheses in order to structure research that can more rapidly advance theory.

Associations between repetitive questioning, resistance to change, temper outbursts and anxiety in Prader-Willi and Fragile-X syndromes

The behavioural phenotypes of Prader-Willi (PWS) and Fragile-X (FraX) syndromes both comprise repetitive behaviours with differences between the profiles. In this study we investigated the context and antecedents to the repetitive behaviours and the association with other behavioural phenotypic characteristics in order to generate testable hypotheses regarding the cause of the behaviours.

The parents or carers of 46 children with PWS (mean age 14.1 years; 20 girls), and 33 boys with FraX (mean age 13.11 years) were interviewed about their children's repetitive behaviour in a semi-structured format.

Children showed negative emotional behaviour (PWS: 87.0%; FraX: 79.4%) and repetitive questions (PWS: 78.3%; FraX: 73.5%) following changes in routine or expectations. Significantly more temper outbursts were reported to follow changes in children with PWS (89.1%) compared with boys with FraX (41.2%) (chi(2) = 20.93; P <0.001). Anxiety that was frequently associated with repetitive and self-injurious behaviours in boys with FraX, followed changes in significantly more boys with FraX (76.5%) compared with children with PWS (6.5%) (chi(2) = 43.19, P <0.001).

On the basis of these reports and existing literature, we hypothesise that decreases in predictability are aversive to children with PWS and FraX. We also hypothesise that these children have a propensity to show a syndrome-related pattern of behaviour (temper outbursts in PWS and displays of anxiety in FraX) when an event in the environment has this aversive property. We hypothesise that questions may be reinforcing to children in their own right by increasing the predictability of the environment. We outline how a specific cognitive deficit in the endophenotypes associated with both PWS and FraX could be investigated as a potential explanation for the hypothesised aversive properties of decreased predictability.

Évaluation neurobiologique des souris spontanément hypertendues : Du vieillissement à la génomique comparative

Le but de cette thèse est premièrement d’évaluer l’effet du vieillissement sur les fonctions psychomotrices des souches de souris sélectionnées génétiquement en fonction de leur tension artérielle (TA); deuxièmement, de localiser les déterminants génétiques des phénotypes psychophysiologiques à partir de souches recombinantes congéniques (RCS). Ces travaux ont mené à la publication de 4 articles. Le premier article décrit l’évaluation des fonctions psychomotrices des souches avec une tension artérielle élevée (HBP), basse (LBP) et normale (NBP). La performance aux épreuves d’exploration, d’habiletés motrices et d’apprentissage spatial, a été mesurée sur deux cohortes âgées respectivement de 12 mois et de trois mois. Indépendamment de l’âge, les HBPs sont hyperactives dans l’open-field (OF), mais pas dans le test d’exploration de trous. Inversement, les LBP explorent moins d’espaces que les NBP et, à trois mois seulement, sont hypoactives dans l’OF. Par ailleurs, les HBPs et les LBP présentent des déficits précoces de coordination motrice et des fonctions visuo-motrices. Le second article concerne l’évaluation longitudinale de la coordination motrice, de l’anxiété et de l’apprentissage spatial des souches HBP, LBP et NBP, à l’âge de deux mois et de 12 mois. Le vieillissement accentue l’hyperactivité des HBPs dans l’OF. Par contre, l’hypoactivité des souris LBP est détectable seulement à l’âge de deux mois. Indépendamment de l’âge, les souris HBP et LBP montrent une perception réduite du danger dans l’épreuve d’anxiété et des dysfonctions visuo-motrices au labyrinthe aquatique. Enfin, des déficits précoces de coordination motrice se manifestent seulement chez les HBPs. Il reste à déterminer si les déficits observés sont liés à des déterminants génétiques indépendants ou secondaires aux altérations de la tension artérielle. Le troisième article présente la comparaison entre les souches consanguines A/J et C57Bl/6J (B6) aux épreuves de l’OF, de la planche à trous, du labyrinthe aquatique et du cintre (coordination motrice). Les B6 explore d’avantage l’OF et la planche à trous. Les B6 sont moins rapides sur le cintre, mais supérieurs aux A/J dans le labyrinthe aquatique, avec une plate-forme invisible ou visible. Ces résultats démontrent l’implication de déterminants génétiques. Cette thèse se termine par un quatrième article sur la localisation des déterminants génétiques de la susceptibilité au stress dans les RCS, dérivées de A/J et B6, et présentant un agencement spécifique de 12.5% du génome. La réactivité émotionnelle est évaluée dans l’OF et le plus-maze; la réponse de stress est mesurée par radio télémétrie de la température interne pendant le stress d’immobilisation (SI) sous diète régulière et riche en sel; l’excrétion des électrolytes urinaires est dosée après 24 heures de diète salée. Les loci les plus significatifs sont situés dans les régions suivantes: de l’émotionalité dans l’OF (Emo1) sur le chr. 1 (LOD=4.6) correspondant à la région homologue impliquée dans la cohorte d’hypertension familiale du Saguenay; de la dopa décarboxylase (ddc) sur le chr. 11 pour l’émergence du plus-maze (LOD=4.7); de la protéine liant l’endotoxine (lbp) sur le chr. 2 pour l’hypothermie initiale en réponse au SI (LOD=4); et de HSP90 sur le chr. 12 pour l’excrétion de Ca++ (LOD=4.6). Des banques de données sont ensuite interrogées pour recenser les polymorphismes des régions régulatrices ou codantes des gènes candidats chez les souches ancestrales A/J et B6, dont les séquences sont disponibles pour le génome entier. Des utilitaires web permettent de dévoiler les changements dans la structure secondaire de l’ARNm, l’interférence avec des microARN ou avec d’autres motifs de liaison. Plusieurs SNPs fonctionnels ont été identifiés pour le QTL du chr. 1, particulièrement dans les éléments de régulation; ceux-ci impliquant des gènes reliés avec les réponses inflammatoire/immunitaire ou avec le système cardiovasculaire. La quantification par la PCR confirme une régulation à la baisse d’atp1a2 dans le cœur et le cerveau des souches susceptibles à l’anxiété. Ces résultats confirment l’intrication des altérations de la susceptibilité au stress et de la régulation de la TA.

A 600 kb deletion syndrome at 16p11.2 leads to energy imbalance and neuropsychiatric disorders

BACKGROUND: The recurrent ~600 kb 16p11.2 BP4-BP5 deletion is among the most frequent known genetic aetiologies of autism spectrum disorder (ASD) and related neurodevelopmental disorders. OBJECTIVE: To define the medical, neuropsychological, and behavioural phenotypes in carriers of this deletion. METHODS: We collected clinical data on 285 deletion carriers and performed detailed evaluations on 72 carriers and 68 intrafamilial non-carrier controls. RESULTS: When compared to intrafamilial controls, full scale intelligence quotient (FSIQ) is two standard deviations lower in carriers, and there is no difference between carriers referred for neurodevelopmental disorders and carriers identified through cascade family testing. Verbal IQ (mean 74) is lower than non-verbal IQ (mean 83) and a majority of carriers require speech therapy. Over 80% of individuals exhibit psychiatric disorders including ASD, which is present in 15% of the paediatric carriers. Increase in head circumference (HC) during infancy is similar to the HC and brain growth patterns observed in idiopathic ASD. Obesity, a major comorbidity present in 50% of the carriers by the age of 7 years, does not correlate with FSIQ or any behavioural trait. Seizures are present in 24% of carriers and occur independently of other symptoms. Malformations are infrequently found, confirming only a few of the previously reported associations. CONCLUSIONS: The 16p11.2 deletion impacts in a quantitative and independent manner FSIQ, behaviour and body mass index, possibly through direct influences on neural circuitry. Although non-specific, these features are clinically significant and reproducible. Lastly, this study demonstrates the necessity of studying large patient cohorts ascertained through multiple methods to characterise the clinical consequences of rare variants involved in common diseases.

Dissecting the complex genetic basis of mate choice

The genetic analysis of mate choice is fraught with difficulties. Males produce complex signals and displays that can consist of a combination of acoustic, visual, chemical and behavioural phenotypes. Furthermore, female preferences for these male traits are notoriously difficult to quantify. During mate choice, genes not only affect the phenotypes of the individual they are in, but can influence the expression of traits in other individuals. How can genetic analyses be conducted to encompass this complexity? Tighter integration of classical quantitative genetic approaches with modern genomic technologies promises to advance our understanding of the complex genetic basis of mate choice.

A Multiple Indicators Multiple Causes (MIMIC) model of Behavioural and Psychological Symptoms in Dementia (BPSD)

Introduction: Although there is evidence for distinct behavioural sub-phenotypes in Alzheimer's disease (AD), their inter-relationships and the effect of clinical variables on their expression have been little investigated.

Developmental, morphological, and behavioural plasticity in the reproductive strategies of stink bugs and their egg parasitoids

L’environnement façonne la physiologie, la morphologie et le comportement des organismes par l’entremise de processus écologiques et évolutifs complexes et multidimensionnels. Le succès reproducteur des animaux est déterminé par la valeur adaptative d’un phénotype dans un environnement en modification constante selon une échelle temporelle d’une à plusieurs générations. De plus, les phénotypes sont façonnés par l’environnement, ce qui entraine des modifications adaptatives des stratégies de reproduction tout en imposant des contraintes. Dans cette thèse, considérant des punaises et leurs parasitoïdes comme organismes modèles, j’ai investigué comment plusieurs types de plasticité peuvent interagir pour influencer la valeur adaptative, et comment la plasticité des stratégies de reproduction répond à plusieurs composantes des changements environnementaux (qualité de l’hôte, radiation ultraviolette, température, invasion biologique). Premièrement, j’ai comparé la réponse comportementale et de traits d’histoire de vie à la variation de taille corporelle chez le parasitoïde Telenomus podisi Ashmead (Hymenoptera : Platygastridae), démontrant que les normes de réaction des comportements étaient plus souvent positives que celles des traits d’histoires de vie. Ensuite, j’ai démontré que la punaise prédatrice Podisus maculiventris Say (Hemiptera : Pentatomidae) peut contrôler la couleur de ses œufs, et que la pigmentation des œufs protège les embryons du rayonnement ultraviolet; une composante d’une stratégie complexe de ponte qui a évoluée en réponse à une multitude de facteurs environnementaux. Puis, j’ai testé comment le stress thermique affectait la dynamique de la mémoire du parasitoïde Trissolcus basalis (Wollaston) (Hymenoptera : Platygastridae) lors de l’apprentissage de la fiabilité des traces chimiques laissées par son hôte. Ces expériences ont révélé que des températures hautes et basses prévenaient l’oubli, affectant ainsi l’allocation du temps passé par les parasitoïdes dans des agrégats d’hôtes contenant des traces chimiques. J’ai aussi développé un cadre théorique général pour classifier les effets de la température sur l’ensemble des aspects comportementaux des ectothermes, distinguant les contraintes des adaptations. Finalement, j’ai testé l’habileté d’un parasitoïde indigène (T. podisi) à exploiter les œufs d’un nouveau ravageur invasif en agriculture, Halyomorpha halys Stål (Hemiptera : Pentatomidae). Les résultats ont montré que T. podisi attaque les œufs de H. halys, mais qu’il ne peut s’y développer, indiquant que le ravageur invasif s’avère un « piège évolutif » pour ce parasitoïde. Cela pourrait indirectement bénéficier aux espèces indigènes de punaises en agissant comme un puits écologique de ressources (œufs) et de temps pour le parasitoïde. Ces résultats ont des implications importantes sur la réponse des insectes, incluant ceux impliqués dans les programmes de lutte biologique, face aux changements environnementaux.

Association between serotonin 5-HT-2C receptor gene (HTR2C) polymorphisms and obesity- and mental health-related phenotypes in a large population-based cohort

OBJECTIVE: Studies have shown that common single-nucleotide polymorphisms (SNPs) in the serotonin 5-HT-2C receptor (HTR2C) are associated with antipsychotic agent-induced weight gain and the development of behavioural and psychological symptoms. We aimed to analyse whether variation in the HTR2C is associated with obesity- and mental health-related phenotypes in a large population-based cohort. METHOD: Six tagSNPs, which capture all common genetic variation in the HTR2C gene, were genotyped in 4978 men and women from the European Prospective Investigation into Cancer (EPIC)-Norfolk study, an ongoing prospective population-based cohort study in the United Kingdom. To confirm borderline significant associations, the -759C/T SNP (rs3813929) was genotyped in the remaining 16 003 individuals from the EPIC-Norfolk study. We assessed social and psychological circumstances using the Health and Life Experiences Questionnaire. Genmod models were used to test associations between the SNPs and the outcomes. Logistic regression was performed to test for association of SNPs with obesity- and mental health- related phenotypes. RESULTS: Of the six HTR2C SNPs, only the T allele of the -759C/T SNP showed borderline significant associations with higher body mass index (BMI) (0.23 kg m(-2); (95% confidence interval (CI): 0.01-0.44); P=0.051) and increased risk of lifetime major depressive disorder (MDD) (Odds ratio (OR): 1.13 (95% CI: 1.01-1.22), P=0.02). The associations between the -759C/T and BMI and lifetime MDD were independent. As associations only achieved borderline significance, we aimed to validate our findings on the -759C/T SNP in the full EPIC-Norfolk cohort (n=20 981). Although the association with BMI remained borderline significant (beta=0.20 kg m(-2); 95% CI: 0.04-0.44, P=0.09), that with lifetime MDD (OR: 1.01; 95% CI: 0.94-1.09, P=0.73) was not replicated. CONCLUSIONS: Our findings suggest that common HTR2C gene variants are unlikely to have a major role in obesity- and mental health-related traits in the general population.

Intraspecific Behavioural Diversity in a Freshwater Zooplankton: A study of the fitness consequences of vertical migration behaviour for distinct morphs of Daphnia pulicaria

The literature on niche separation and coexistence between species is large, but there is widespread variation in behavioural strategy between individuals of the same species that has received much less attention. Understanding what maintains this diversity is important because intraspecific behavioural diversity can affect population dynamics and community interactions. Multiple behavioural strategies can arise either as phenotype-dependent ‘conditional strategies’, where phenotypic variation causes individuals to adopt different strategies for optimizing fitness, or as internally-independent ‘alternative strategies’, where multiple fitness peaks exist for individuals and strategic ‘choice’ remains plastic. Though intraspecific variation in stable phenotypes is known to maintain intraspecific behavioural diversity through conditional strategies, when internal conditions are highly plastic or reversible, it is not clear whether individual behaviours are maintained as conditional strategies, or as alternative strategies of equal fitness. In this study, I combine an observational and experimental approach to identify the likely mechanisms maintaining behavioural diversity between hemoglobin-rich and hemoglobin-poor morphs in a natural population of Daphnia pulicaria. In Round Lake, individuals with low hemoglobin migrate daily from the hypolimnion to the epilimnion, whereas individuals with high hemoglobin remain in the hypolimnion. Using high-resolution depth and time sampling, I discovered behavioural diversity both within and among hemoglobin phenotypes. I tested the role of hemoglobin phenotype in maintaining behavioural diversity using automated migration robots that move individuals across the natural environmental gradients in the lake. By measuring the fitness of each morph undergoing either a natural migration behaviour, or the migration of the opposite morph, I found that the fitness of hemoglobin rich and poor morphs in their natural behaviour does not differ, but that Hb-rich individuals can obtain equal fitness from either behaviour, while Hb-poor morphs suffer substantial drops in survivorship in the alternate migration behaviour. Thus, migration behaviour in this system exists as a conditional strategy for some individuals, and as alternative strategies of equal fitness for others. The results of this study suggest that individual limits in the expression of highly flexible internal conditions can reinforce intraspecific behavioural diversity. Few studies have measured the fitness consequences of switching migration strategies and this study provides a rare example in the field.