Resting frontal EEG asymmetry in adolescents with major depression: Impact of disease state and comorbid anxiety disorder.

OBJECTIVE: Greater relative right- than left-frontal cortical activity has been frequently found in adults with major depression (MD). As the few studies in adolescents with MD have been inconclusive, the aim of this study was to assess frontal alpha asymmetry (FAA) in an adolescent sample with MD whilst taking into account possible confounding variables such as disease state and comorbid anxiety disorder. METHODS: An 8-minute resting frontal EEG was assessed in 34 healthy controls (HCs), 16 adolescents with MD in remission without comorbid anxiety disorder (rMDa-), 22 adolescents with acute depression without comorbid anxiety disorder (MDa-), and 23 adolescents with acute depression and comorbid anxiety disorder (MDa+). Alpha power was analyzed over corresponding frontal Regions of Interests. RESULTS: Compared to HCs, MDa+ adolescents demonstrated more left- than right-sided EEG alpha power, indicating greater right-than left-frontal cortical activity. No other group differences emerged. CONCLUSIONS: The results suggest that greater relative right-frontal cortical activity in adolescent MD is not a result of disease state but can be attributed to comorbid anxiety disorder. SIGNIFICANCE: Results suggest that FAA is not linked to adolescent depression per se and highlight the importance of considering comorbid disorders when examining asymmetry patterns in adolescent MD.

Resting frontal EEG asymmetry patterns in adolescents with and without major depression.

More right-sided frontal brain resting activity has been postulated to be a correlate of major depression in adults. In children and adolescents, more right-sided activity (as indicated by more left-sided alpha activity) seems to be associated with psychosocial risk factors. However, an association of frontal asymmetry and manifest unipolar depression has not been shown in adolescents so far. We analyzed frontal asymmetry in 20 adolescents (12-17 years) with unipolar depression (12 with first episode, 8 with recurrent depression) and 31 healthy age-matched controls. We found significantly less left-sided alpha power over frontal Regions of Interest in patients, while controls exhibited no asymmetry. In the control group, more left-sided frontal alpha correlated with higher depression scores, which was not observed in the patient group. Our results suggest developmental effects on frontal asymmetry, and prompt further investigations in adolescents to clarify the role of frontal asymmetry in adolescent major depression.

Not later, but longer: sleep, chronotype and light exposure in adolescents with remitted depression compared to healthy controls.

The relationship between sleep and adolescent depression is much discussed, but still not fully understood. One important sleep variable is self-selected sleep timing, which is also referred to as chronotype. Chronotype is mostly regulated by the circadian clock that synchronises the internal time of the body with the external light dark cycle. A late chronotype as well as a misalignment between internal time and external time such as social jetlag has been shown to be associated with depressive symptoms in adults. In this study, we investigated whether adolescents with remitted depression differ from healthy controls in terms of chronotype, social jetlag and other sleep-related variables. For this purpose, we assessed chronotype and social jetlag with the Munich ChronoType Questionnaire (MCTQ), subjective sleep quality with the Pittsburgh Sleep Quality Index (PSQI) and used continuous wrist-actimetry over 31 consecutive days to determine objective sleep timing. Given the potentially mediating effect of light on chronotype and depressive symptoms, we measured light exposure with a light sensor on the actimeter. In our sample, adolescents with remitted depression showed similar chronotypes and similar amounts of social jetlag compared to controls. However, patients with remitted depression slept significantly longer on work-free days and reported a worse subjective sleep quality than controls. Additionally, light exposure in remitted patients was significantly higher, but this finding was mediated by living in a rural environment. These findings indicate that chronotype might be modified during remission, which should be further investigated in longitudinal studies.

[Chronotype and depression in adolescents ­ a review]. / Chronotyp und Depression bei Jugendlichen ­ ein Review.

Many patients with depressive disorders experience symptoms in relation to sleep behavior and daily rhythmicity. However, the multifaceted associations between sleep, depression and circadian rhythms are not fully understood. During the past years, the concept of chronotypehas become increasingly popular in research. The Munich Chronotype Questionnaire (MCTQ) derives chronotype from sleep timing on work-free days and therefore represents a biological measure for the circadian clock, whereas the Morningness-Eveningness-Questionnaire(MEQ) assesses chronotype as a subjective preference for different activities at specific times of day. Chronotype changes with age, with adolescents and young adults being especially late types. We conducted a systematic literature research and identified studies that explore the association between chronotype (MEQ, MCTQ) and depressive symptoms or depressive disorders. Most of the studies showed an association between a late chronotype and depressive symptomatology. However, it is still unclear what is cause and effect. We propose a bidirectional relationship: On the one hand, due to reduced social and physical activity, depressed patients get less daylight which causes their chronotype to delay. On the other hand, a discrepancy between internal time (directed by the circadian clock) and external time (such as early school- or works tarting times) can cause problems

Right-lateralization of N2-amplitudes in depressive adolescents: an emotional go/no-go study.

BACKGROUND: Recent studies have proposed the process of emotion regulation as a promising target to study the neurophysiological basis of adolescent depression. Emotion regulation has repeatedly been studied with emotional go/no-go paradigms. To date, no study has examined if the left-frontal hypoactivation associated with depression generalizes to active tasks. The aim of this study was therefore to investigate the hemispheric asymmetry of the N2 component in depressed adolescents in an emotion regulation paradigm. METHODS: Twenty-four adolescents diagnosed with major depression (age 11-18) and 30 healthy controls (age 11-18) performed two emotional go/no-go tasks exhibiting negative faces as go trials and positive faces as no-go trials and vice versa. RESULTS: On the behavioral level, no significant group differences emerged. On the neural level, we found a more right-lateralized N2-amplitude in depressed subjects, while it was more left-lateralized in controls. Furthermore, both groups showed a less negative N2-amplitude to positive no-go stimuli. CONCLUSION: This study provides strong support for a general left-frontal hypoactivity in adolescent depression, which also applies to active emotional go/no-go paradigms. Furthermore, the less negative N2 to positive stimuli is consistent with a generally enhanced impulsivity of adolescents toward appetitive stimuli, which is possibly the base of the differential clinical pattern of adolescent in contrast to adult depression.

"Glass fairies" and "bone children": adolescents and young adults with anorexia nervosa show positive reactions towards extremely emaciated body pictures measured by the startle reflex paradigm.

In this study, we investigated the emotional processing of extremely emaciated body cues in adolescents and young adults with (n = 36) and without (n = 36) anorexia nervosa (AN), introducing a new picture type, which was taken from websites that promote extreme thinness and is targeted specifically at adolescents interested in extreme thinness. A startle reflex paradigm was used for implicit reactions, while a self-assessment instrument was used for subjective responses. We found a significant group difference with a startle inhibition (appetitive response) among the patients and a startle potentiation (aversive response) among the controls, whereas no such difference for subjective measures was found. The results are in contrast to previous studies, which proposed a general failure to activate the appetitive motivational system in AN, but in keeping with findings from other addictions, where the same response pattern has been found. Implications for prevention and therapy are discussed.

Formation of the retinotectal projection requires Esrom, an ortholog of PAM (protein associated with Myc).

Visual system development is dependent on correct interpretation of cues that direct growth cone migration and axon branching. Mutations in the zebrafish esrom gene disrupt bundling and targeting of retinal axons, and also cause ectopic arborization. By positional cloning, we establish that esrom encodes a very large protein orthologous to PAM (protein associated with Myc)/Highwire/RPM-1. Unlike motoneurons in Drosophila highwire mutants, retinal axons in esrom mutants do not arborize excessively, indicating that Esrom has different functions in the vertebrate visual system. We show here that Esrom has E3 ligase activity and modulates the amount of phosphorylated Tuberin, a tumor suppressor, in growth cones. These data identify a mediator of signal transduction in retinal growth cones, which is required for topographic map formation.

Chemically controlled formation of a DNA/calcium phosphate coprecipitate: application for transfection of mature hippocampal neurons.

Numerous methods exist for transfecting postmitotic neurons, for example, DNA/calcium phosphate coprecipitation, cationic lipids, viruses, and physical methods such as microinjection, electroporation, and biolistics. Most methods, however, are either toxic to the cell, yield only poor transfection efficiencies, or cells have to be electroporated before plating. In this article, we present a standardized and fast transfection method using DNA/calcium phosphate coprecipitates that efficiently transfer DNA into mature, postmitotic hippocampal neurons. Shifting to CO(2)-independent media with a well-defined pH allows for the tight control of the coprecipitate formation and for adjusting the transfection parameters for the individual DNA plasmid used. The two critical parameters for reproducible and efficient transfections are: the precise pH during crystal formation, and the incubation time of the cells with the coprecipitate. This improved procedure now enables biochemical approaches. By transfecting a dominant-positive Ras mutant, we activate the Erk/MAP kinase signal transduction pathway. Furthermore, using a siRNA plasmid directed against MAP2, the level of an endogenously expressed protein is down-regulated upon transfection. These two approaches demonstrate that the presented transient transfection method can now be used to address questions on a biochemical level in hippocampal neurons.

The brain-specific double-stranded RNA-binding protein Staufen2: nucleolar accumulation and isoform-specific exportin-5-dependent export.

The mammalian double-stranded RNA-binding proteins Staufen (Stau1 and Stau2) are involved in RNA localization in polarized neurons. In contrast to the more ubiquitously expressed Stau1, Stau2 is mainly expressed in the nervous system. In Drosophila, the third double-stranded RNA-binding domain (RBD3) of Staufen is essential for RNA interaction. When conserved amino acids within the RBD3 of Stau2 were mutated to render Stau2 defective for RNA binding, the mutant Stau2 proteins accumulate predominantly in the nucleolus. This is in contrast to wild type Stau2 that mostly localizes in the cytosol. The nuclear import is dependent on a nuclear localization signal in close proximity to the RBD3. The nuclear export of Stau2 is not dependent on CRM1 but rather on Exportin-5. We show that Exportin-5 interacts with the RBD3 of wild type Stau2 in an RNA-dependent manner in vitro but not with mutant Stau2. When Exportin-5 is down-regulated by RNA interference, only the largest isoform of Stau2 (Stau2(62)) preferentially accumulates in the nucleolus. It is tempting to speculate that Stau2(62) binds RNA in the nucleus and assembles into ribonucleoparticles, which are then exported via the Exportin-5 pathway to their final destination.

EphrinB2a in the zebrafish retinotectal system.

Members of the Eph-B family of receptors tyrosine kinase and their transmembrane ligands have been implicated in dorsoventral patterning of the vertebrate retinotectal projection. In the zebrafish retinotectal system, however, ephrinB2a is expressed strongly in the posterior tectum, in tectal neurons that form physical contacts with retinal ganglion cell (RGC) axons. In the gnarled mutant, where tectal neurons form ectopically in the pretectum, RGC axons stall before entering the tectum, or else are misrouted or branch aberrantly in the tectal neuropil. Ectopic expression of ephrinB2a in the anterior midbrain of wild-type embryos, with the aid of baculovirus, also inhibits RGC axon entry into the tectum. In vitro, zebrafish RGC axons are repelled by stripes of purified ephrinB2a. It is proposed that ephrinB2a may signal a subpopulation of RGC axons that they have reached their target neurons in the tectum.

Coupling the iron-responsive element to GFP--an inducible system to study translation in a single living cell.

Local protein synthesis in a cell represents an elegant mechanism to achieve important biological phenomena such as cell migration, body axis formation during embryonic development and establishment of cell polarity. A prerequisite to studying translation in a restricted cellular compartment is the ability to unambiguously discriminate between proteins that arise through local protein synthesis and those that reach the site of interest by diffusion or transport. To tackle this problem, we set up a green fluorescent protein (GFP)-based reporter system that allows one to uncouple the translation of reporter gene mRNA from its subcellular localization. The system is based on the iron-responsive element, which regulates the translation of both endogenous ferritin and transferrin transcripts in response to changes in iron concentration. Translation of the reporter messenger RNA (mRNA) is thus dependent on iron in the medium; both its transcription and localization, however, are unaffected. Known targeting sequences can be used to direct the mRNA transcript to a subcellular compartment of interest. For instance, the full-length 3'-untranslated region of calcium/calmodulin-dependent protein kinase IIalpha mRNA can be added to the construct, after the stop codon of the GFP sequence, to selectively target the transcript into the dendrites of transiently transfected hippocampal neurons. This novel fluorescent assay will allow us to address a number of important biological questions in living mammalian cells.

Barentsz, a new component of the Staufen-containing ribonucleoprotein particles in mammalian cells, interacts with Staufen in an RNA-dependent manner.

Staufen1, the mammalian homolog of Drosophila Staufen, assembles into ribonucleoprotein particles (RNPs), which are thought to transport and localize RNA into dendrites of mature hippocampal neurons. We therefore investigated whether additional components of the RNA localization complex besides Staufen are conserved. One candidate is the mammalian homolog of Drosophila Barentsz (Btz), which is essential for the localization of oskar mRNA to the posterior pole of the Drosophila oocyte and is a component of the oskar RNA localization complex along with Staufen. In this study, we report the characterization of mammalian Btz, which behaves like a nucleocytoplasmic shuttling protein. When expressed in the Drosophila egg chamber, mammalian Btz is still able to interact with Drosophila Staufen and reach the posterior pole in the wild-type oocyte, but does not rescue the btz mutant phenotype. Most interestingly, we show by immunoprecipitation assays that Btz interacts with mammalian Staufen in an RNA-dependent manner through a conserved domain, which encompasses the region of homology to the Drosophila Btz protein and contains a novel conserved motif. One candidate for an RNA that mediates this interaction is the dendritically localized brain cytoplasmic 1 transcript. In addition, Btz and Staufen1 colocalize within particles in the cell body and, to a more variable extent, in dendrites of mature hippocampal neurons. Together, our data suggest that the mRNA transport machinery is conserved during evolution, and that mammalian Btz is an additional component of the dendritic RNPs in hippocampal neurons.

A GFP-based system to uncouple mRNA transport from translation in a single living neuron.

An inducible fluorescent system based on GFP is presented that allows for the uncoupling of dendritic mRNA transport from subsequent protein synthesis at the single cell level. The iron-responsive element (IRE) derived from ferritin mRNA in the 5'-UTR of the GFP reporter mRNA renders translation of its mRNA dependent on iron. The addition of the full-length 3'-UTR of the Ca(2+)/calmodulin-dependent protein kinase II alpha (CaMKIIalpha) after the stop codon of the GFP reading frame targets the reporter mRNA to dendrites of transfected fully polarized hippocampal neurons. As we show by time-lapse videomicroscopy, iron specifically turns on GFP reporter protein synthesis in a single transfected hippocampal neuron. We investigate whether GFP expression is affected--in addition to iron--by synaptic activity. Interestingly, synaptic activity has a clear stimulatory effect. Most importantly, however, this activity-dependent protein synthesis is critically dependent on the presence of the full-length 3'-UTR of CaMKIIalpha confirming that this sequence contains translational activation signals. The IRE-based system represents a new convenient tool to study local protein synthesis in mammalian cells where mRNA localization to a specific intracellular compartment occurs.