Melanie P. Leussis

The ANK3 Bipolar Disorder Gene Regulates Psychiatric-Related Behaviors That Are Modulated by Lithium and Stress

BACKGROUND Ankyrin 3 (ANK3) has been strongly implicated as a risk gene for bipolar disorder (BD) by recent genome-wide association studies of patient populations. However, the genetic variants of ANK3 contributing to BD risk and their pathological function are unknown. METHODS To gain insight into the potential disease relevance of ANK3, we examined the function of mouse Ank3 in the regulation of psychiatric-related behaviors using genetic, neurobiological, pharmacological, and gene-environment interaction (G×E) approaches. Ank3 expression was reduced in mouse brain either by viral-mediated RNA interference or through disruption of brain-specific Ank3 in a heterozygous knockout mouse. RESULTS RNA interference of Ank3 in hippocampus dentate gyrus induced a highly specific and consistent phenotype marked by decreased anxiety-related behaviors and increased activity during the light phase, which were attenuated by chronic treatment with the mood stabilizer lithium. Similar behavioral alterations of reduced anxiety and increased motivation for reward were also exhibited by Ank3+/- heterozygous mice compared with wild-type Ank3+/+ mice. Remarkably, the behavioral traits of Ank3+/- mice transitioned to depression-related features after chronic stress, a trigger of mood episodes in BD. Ank3+/- mice also exhibited elevated serum corticosterone, suggesting that reduced Ank3 expression is associated with elevated stress reactivity. CONCLUSIONS This study defines a new role for Ank3 in the regulation of psychiatric-related behaviors and stress reactivity that lends support for its involvement in BD and establishes a general framework for determining the disease relevance of genes implicated by patient genome-wide association studies.

Abnormal behavioral and neurotrophic development in the younger sibling receiving less maternal care in a communal nursing paradigm in rats

Maternal behavior in rodents has been proposed to vary as a function of the external environment and, in turn, adjust offsprings stress and fear responses. Early handling (brief periods of maternal separation during the first two weeks of life) studies and analyses of spontaneously high-caring rat mothers converge to indicate that increased levels of maternal care may reduce offspring emotionality in adulthood. However, the hypothesis that environment-dependent reduction in maternal care correlates with increased offspring vulnerability to pathology has been scarcely investigated. To test this hypothesis we studied maternal care and offspring development in young, adolescent and young-adult Sprague-Dawley rats reared in a communal nursing situation, characterized by two dams delivering their offspring four days apart and communally caring for them until weaning. We show that dams of the first-born litter show increased aggression towards the pregnant female and that offspring belonging to the second-born litter receive less maternal care compared to older cage-mates. Additionally, second-born rats show increased anxiety-related behavior in a plus-maze test in adolescence and adulthood and abnormal developmental trajectories in terms of social interaction and BDNF levels in the amygdala and hippocampus compared to both the first-born litter and to animal facility reared controls. This is the first indication that adverse environments, not requiring experimenter handling, may reduce maternal care and in turn increase offsprings emotionality and modify social behavior and BDNF developmental trajectories.

Depressive-like behavior in adolescents after maternal separation: sex differences, controllability, and GABA.

Exposure to adversity during development is an identified risk factor for depression later in life. In humans, early adversity accelerates the onset of depressive symptoms, which manifest during adolescence. Animal studies have used maternal separation as a model of early adversity to produce adult depressive-like behaviors, but have yet to examine these behaviors during adolescence. Moreover, the nature of depressive-like behaviors has not been well characterized in this model. Here, we used the triadic model of learned helplessness to understand controllability, helplessness, and motivational factors following maternal separation in male and female adolescent rats. We found sex-dependent changes in the effects of separation, with males demonstrating loss of controllability in an escapable shock condition, whereas females demonstrated motivational impairment in a no-shock condition. The effect, however, did not endure as adult females were no longer helpless. Reductions in parvalbumin, a GABAergic marker, in the prefrontal cortex of separated subjects relative to age-matched controls were evident and paralleled depressive-like behavior. Understanding the risk factors for depression, the nature of depressive-like behaviors, and their unique sex dependency may ultimately provide insight into improved treatments.

Ankyrin 3: genetic association with bipolar disorder and relevance to disease pathophysiology

Bipolar disorder (BD) is a multi-factorial disorder caused by genetic and environmental influences. It has a large genetic component, with heritability estimated between 59-93%. Recent genome-wide association studies (GWAS) using large BD patient populations have identified a number of genes with strong statistical evidence for association with susceptibility for BD. Among the most significant and replicated genes is ankyrin 3 (ANK3), a large gene that encodes multiple isoforms of the ankyrin G protein. This article reviews the current evidence for genetic association of ANK3 with BD, followed by a comprehensive overview of the known biology of the ankyrin G protein, focusing on its neural functions and their potential relevance to BD. Ankyrin G is a scaffold protein that is known to have many essential functions in the brain, although the mechanism by which it contributes to BD is unknown. These functions include organizational roles for subcellular domains in neurons including the axon initial segment and nodes of Ranvier, through which ankyrin G orchestrates the localization of key ion channels and GABAergic presynaptic terminals, as well as creating a diffusion barrier that limits transport into the axon and helps define axo-dendritic polarity. Ankyrin G is postulated to have similar structural and organizational roles at synaptic terminals. Finally, ankyrin G is implicated in both neurogenesis and neuroprotection. ANK3 and other BD risk genes participate in some of the same biological pathways and neural processes that highlight several mechanisms by which they may contribute to BD pathophysiology. Biological investigation in cellular and animal model systems will be critical for elucidating the mechanism through which ANK3 confers risk of BD. This knowledge is expected to lead to a better understanding of the brain abnormalities contributing to BD symptoms, and to potentially identify new targets for treatment and intervention approaches.

The ankyrin-3 gene is associated with posttraumatic stress disorder and externalizing comorbidity.

BACKGROUND The ankyrin 3 gene (ANK3) produces the ankyrin G protein that plays an integral role in regulating neuronal activity. Previous studies have linked ANK3 to bipolar disorder and schizophrenia. A recent mouse study suggests that ANK3 may regulate behavioral disinhibition and stress reactivity. This led us to hypothesize that ANK3 might also be associated with stress-related psychopathology such as posttraumatic stress disorder (PTSD), as well as disorders of the externalizing spectrum such as antisocial personality disorder and substance-related disorders that are etiologically linked to impulsivity and temperamental disinhibition. METHODS We examined the possibility of association between ANK3 SNPs and both PTSD and externalizing (defined by a factor score representing a composite of adult antisociality and substance abuse) in a cohort of white non-Hispanic combat veterans and their intimate partners (n=554). Initially, we focused on rs9804190-a SNP previously reported to be associated with bipolar disorder, schizophrenia, and ankyrin G expression in brain. Then we examined 358 additional ANK3 SNPs utilizing a multiple-testing correction. RESULTS rs9804190 was associated with both externalizing and PTSD (p=0.028 and p=0.042 respectively). Analysis of other ANK3 SNPs identified several that were more strongly associated with either trait. The most significant association with externalizing was observed at rs1049862 (p=0.00040, pcorrected=0.60). The most significant association with PTSD (p=0.00060, pcorrected=0.045) was found with three SNPs in complete linkage disequilibrium (LD)-rs28932171, rs11599164, and rs17208576. CONCLUSIONS These findings support a role of ANK3 in risk of stress-related and externalizing disorders, beyond its previous associations with bipolar disorder and schizophrenia.

Juvenile Methylphenidate Exposure and Factors That Influence Incentive Processing

Methylphenidate (MPH) is one of the few psychotropic agents approved for use in pediatric populations, underscoring the importance of elucidating any long-term consequences following exposure to this agent. Here, we examined the influence of several variables (i.e. age of assessment, age of exposure, sex, route of administration) on the effect of chronic low-dose MPH (2 mg/kg, twice daily) exposure on place conditioning to cocaine. Juvenile exposure to MPH, but not later exposure, resulted in aversions to cocaine-paired environments when assessed in young adult male rats, but not those entering adolescence. Juvenile MPH enhanced place preferences for cocaine-paired environments in female adolescent rats. The route of administration (i.p. injection or oral ingestion) did not produce enduring differential effects on behavior, and D-MPH was confirmed as the active enantiomer. These observations add to the growing literature on the enduring effects of MPH exposure, and highlight the need for more research in females.

Seizure susceptibility and locus ceruleus activation are reduced following environmental enrichment in an animal model of epilepsy

Alterations in the complexity of social and physical housing environments modulate seizure susceptibility in animal models of epilepsy. The studies described here tested the hypothesis that environmental enrichment would delay seizure onset in the epileptic (El) mouse. Neural activation measured via cFos expression, accumulation of the stress neuropeptide corticotropin-releasing factor (CRF), and behavioral seizure susceptibility were quantified in El mice to better understand the mechanisms of ictogenesis. Enrichment housing of El mice from Postnatal Days 21 to 49 produced a 100% decrease in seizure susceptibility relative to El controls. cFos expression increased in the primary motor cortex, locus ceruleus, and hippocampus of El mice relative to ddY controls, an effect attenuated by enrichment housing. CRF levels were elevated by enrichment in the hippocampus of ddY mice only. This study provides evidence that enrichment housing delays the onset of seizure susceptibility in El mice while altering the neuronal and stress-related responses in seizure-associated regions of the El brain.

Genomic survey of prepulse inhibition in mouse chromosome substitution strains

Prepulse inhibition (PPI) is a measure of sensorimotor gating, a pre‐attentional inhibitory brain mechanism that filters extraneous stimuli. Prepulse inhibition is correlated with measures of cognition and executive functioning, and is considered an endophenotype of schizophrenia and other psychiatric illnesses in which patients show PPI impairments. As a first step toward identifying genes that regulate PPI, we performed a quantitative trait locus (QTL) screen of PPI phenotypes in a panel of mouse chromosome substitution strains (CSSs). We identified five CSSs with altered PPI compared with the host C57BL/6J strain: CSS‐4 exhibited decreased PPI, whereas CSS‐10, ‐11, ‐16 and ‐Y exhibited higher PPI compared with C57BL/6J. These data indicate that A/J chromosomes 4, 10, 11, 16 and Y harbor at least one QTL region that modulates PPI in these CSSs. Quantitative trait loci for the acoustic startle response were identified on seven chromosomes. Like PPI, habituation of the startle response is also disrupted in schizophrenia, and in the present study CSS‐7 and ‐8 exhibited deficits in startle habituation. Linkage analysis of an F2 intercross identified a highly significant QTL for PPI on chromosome 11 between positions 101.5 and 114.4 Mb (peak LOD = 4.54). Future studies will map the specific genes contributing to these QTLs using congenic strains and other genomic approaches. Identification of genes that modulate PPI will provide insight into the neural mechanisms underlying sensorimotor gating, as well as the psychopathology of disorders characterized by gating deficits.

Quality of rearing guides expression of behavioral and neural seizure phenotypes in EL mice

The present studies employed behavioral and neural markers of seizure-related plasticity to examine the relative contributions of genetic predisposition versus rearing environment in generating adult phenotypes in EL mice, a stress-induced animal model of epilepsy. Early environment was manipulated by cross-fostering pups of the EL strain to a seizure-resistant CD-1 control strain of mouse. The impact of changes in rearing quality on growth,exploratory and stress-reactivity phenotypes were examined, with a focus on the role of maternal care in shaping seizure susceptibility and neural cF os activation. Improvement in maternal care imposed by replacing biological EL dams with foster CD-1 mothers was sufficient to decrease pup mortality, to increase body weight gain (+0.1 g/day) and to delay the onset of seizure susceptibility in EL offspring beyond post-natal day 80–90. Moreover,hypoactivity in hippocampus and cortex among EL offspring cross-fostered to EL, but not CD-1 control, dams suggests that changes in rearing environment were accompanied by enduring changes in brain plasticity. Thus, neural and behavioral phenotypes of EL mice are dependent upon post-partum maternal care which if systematically enhanced can postpone seizure expression.

Routine tail suspension husbandry facilitates onset of seizure susceptibility in EL mice.

Summary:  Purpose: Tail suspension can elicit seizures in susceptible EL mice, a model of idiopathic, multifactorial epilepsy. Further, repeated tail suspension hastens the lifetime onset of seizure susceptibility in these mice. The present study tested the hypothesis that curtailing human handling during development would delay the onset of seizure susceptibility relative to EL mice handled regularly by using tail suspension for standard laboratory husbandry.