Multiple inducers and novel roles of autoantibodies against the obligatory NMDAR subunit NR1: a translational study from chronic life stress to brain injury.

Circulating autoantibodies (AB) of different immunoglobulin classes (IgM, IgA, and IgG), directed against the obligatory N-methyl-D-aspartate-receptor subunit NR1 (NMDAR1-AB), belong to the mammalian autoimmune repertoire, and appear with age-dependently high seroprevalence across health and disease. Upon access to the brain, they can exert NMDAR-antagonistic/ketamine-like actions. Still unanswered key questions, addressed here, are conditions of NMDAR1-AB formation/boosting, intraindividual persistence/course in serum over time, and (patho)physiological significance of NMDAR1-AB in modulating neuropsychiatric phenotypes. We demonstrate in a translational fashion from mouse to human that (1) serum NMDAR1-AB fluctuate upon long-term observation, independent of blood-brain barrier (BBB) perturbation; (2) a standardized small brain lesion in juvenile mice leads to increased NMDAR1-AB seroprevalence (IgM + IgG), together with enhanced Ig-class diversity; (3) CTLA4 (immune-checkpoint) genotypes, previously found associated with autoimmune disease, predispose to serum NMDAR1-AB in humans; (4) finally, pursuing our prior findings of an early increase in NMDAR1-AB seroprevalence in human migrants, which implicated chronic life stress as inducer, we independently replicate these results with prospectively recruited refugee minors. Most importantly, we here provide the first experimental evidence in mice of chronic life stress promoting serum NMDAR1-AB (IgA). Strikingly, stress-induced depressive-like behavior in mice and depression/anxiety in humans are reduced in NMDAR1-AB carriers with compromised BBB where NMDAR1-AB can readily reach the brain. To conclude, NMDAR1-AB may have a role as endogenous NMDAR antagonists, formed or boosted under various circumstances, ranging from genetic predisposition to, e.g., tumors, infection, brain injury, and stress, altogether increasing over lifetime, and exerting a spectrum of possible effects, also including beneficial functions.

Uncoupling the widespread occurrence of anti-NMDAR1 autoantibodies from neuropsychiatric disease in a novel autoimmune model.

Autoantibodies of the IgG class against N-methyl-D-aspartate-receptor subunit-NR1 (NMDAR1-AB) were considered pathognomonic for anti-NMDAR encephalitis. This view has been challenged by the age-dependent seroprevalence (up to >20%) of functional NMDAR1-AB of all immunoglobulin classes found in >5000 individuals, healthy or affected by different diseases. These findings question a merely encephalitogenic role of NMDAR1-AB. Here, we show that NMDAR1-AB belong to the normal autoimmune repertoire of dogs, cats, rats, mice, baboons, and rhesus macaques, and are functional in the NMDAR1 internalization assay based on human IPSC-derived cortical neurons. The age dependence of seroprevalence is lost in nonhuman primates in captivity and in human migrants, raising the intriguing possibility that chronic life stress may be related to NMDAR1-AB formation, predominantly of the IgA class. Active immunization of ApoE-/- and ApoE+/+ mice against four peptides of the extracellular NMDAR1 domain or ovalbumin (control) leads to high circulating levels of specific AB. After 4 weeks, the endogenously formed NMDAR1-AB (IgG) induce psychosis-like symptoms upon MK-801 challenge in ApoE-/- mice, characterized by an open blood-brain barrier, but not in their ApoE+/+ littermates, which are indistinguishable from ovalbumin controls. Importantly, NMDAR1-AB do not induce any sign of inflammation in the brain. Immunohistochemical staining for microglial activation markers and T lymphocytes in the hippocampus yields comparable results in ApoE-/- and ApoE+/+ mice, irrespective of immunization against NMDAR1 or ovalbumin. These data suggest that NMDAR1-AB of the IgG class shape behavioral phenotypes upon access to the brain but do not cause brain inflammation on their own.

Dissociation of Pe and ERN/Ne in the conscious recognition of an error.

We evaluated the relationship between conscious awareness and the ERN/Ne and Pe in a digit entering task. On each trial, participants rated the accuracy of their responses on a three-point scale (incorrect, unsure, correct). The ERN/Ne was present on incorrect trials judged as incorrect. The Pe was evident on the same trials but also on correct and incorrect trials judged as unsure. We propose that the ERN/Ne occurs when there is an incorrect execution of a correct motor plan and the representation of the correct response is available for comparison with the actual response. The mismatch information that results from this comparison can be transferred to the Pe process and conscious awareness. However, the Pe process and conscious awareness do not only depend on this transfer of information from the ERN/Ne process. The Pe also occurs when there is uncertainty about the correctness of the motor plan, whether or not the plan is, in fact, correct.

Dissociation of acquisition and expression of fear conditioned responses under working memory load.

Previously, it has been suggested that high working memory load (WML) prevents associative learning and, thus, conditioned responses (CRs) during differential-delay fear conditioning. However, previous studies did not distinguish between the effects of WML on the acquisition versus the expression of the CRs. In the present study, we reinvestigated this issue by manipulating WML during either acquisition or postconditioning and showed that the absence of a differential CR under high WML does not indicate the absence of the acquisition of this CR. Thus, a significant CR was observed during a task-free postconditioning phase, and there was no CR during the preceding acquisition phase with high WML. Conversely, a normally acquired CR was impaired by high WML during the postconditioning phase. The results suggest that high WML affects the expression of the CR. If they are not experimentally separated, the effects of WML on the acquisition and expression of the CR may occur together, and the findings may be erroneously interpreted as indicating a lack of associative learning.

Why humans deviate from rational choice.

Rational choice theory predicts that humans always optimize the expected utility of options when making decisions. However, in decision-making games, humans often punish their opponents even when doing so reduces their own reward. We used the Ultimatum and Dictator games to examine the affective correlates of decision-making. We show that the feedback negativity, an event-related brain potential that originates in the anterior cingulate cortex that has been related to reinforcement learning, predicts the decision to reject unfair offers in the Ultimatum game. Furthermore, the decision to reject is positively related to more negative emotional reactions and to increased autonomic nervous system activity. These findings support the idea that subjective emotional markers guide decision-making and that the anterior cingulate cortex integrates instances of reinforcement and punishment to provide such affective markers.

Hypersensitivity to reward in problem gamblers.

BACKGROUND: Recent research has begun to examine the neurophysiologic basis of pathological gambling. However, direct evidence of a behavioral deficit and an accompanying neurofunctional deviation in a realistic gambling context such as Black Jack has not yet been reported. METHODS: Electroencephalogram was recorded while 20 problem gamblers and 21 control participants played a computerized version of Black Jack. Participants were asked to decide at point scores between 11 and 21 whether they wanted to take another card ("hit") to arrive closer to 21 than the opponent (simulated by computer) or not to take another card ("sit") to avoid going over 21 ("bust"). RESULTS: At a critical point score of 16, problem gamblers decided more often to hit despite losses due to a bust on the preceding trial, whereas control participants decided more often to sit under these conditions. Furthermore, problem gamblers showed more reward-related positive amplitudes in the event-related brain potential than control participants after successful hit decisions at 16. CONCLUSIONS: Here we provide experimental evidence for high-risk taking behavior in gamblers and its correlate in event-related brain potentials. Our results suggest that high-risk-taking behavior in problem gamblers is associated with an increased reward-related neural response to infrequent successes of this behavior.

Decision-making under risk: an fMRI study.

Recent research has focused on decision-making under risk and its neural bases. Two kinds of bad decisions under risk may be defined: too risky decisions and too cautious decisions. Here we show that suboptimal decisions of both kinds lead to increased activity in the anterior cingulate cortex in a Blackjack gambling task. Moreover, this increased activity is related to the avoidance of the negatively evaluated decision under risk. These findings complement other results suggesting an important role of the dorsal anterior cingulate cortex in reward-based decision-making and conflict resolution.

An electrophysiological analysis of coaching in Blackjack.

Recent research has demonstrated that a negative deflection in the event-related potential (ERP) that is usually elicited by errors, negative performance feedback, and monetary losses, and which has been associated with response monitoring and reinforcement learning, is also present when we observe others. In the present study we aimed to extend these findings to the domain of coaching behavior. In many contexts of human social life, advice is given by experts to novices, e.g., teachers or parents to scholars or children. However, their advice is sometimes rejected. Here we show that a rejection of one's advice elicits the same negative potential as when one receives negative feedback about one's own behavior.

Drive for thinness and attention toward specific body parts in a nonclinical sample.

OBJECTIVE: To examine the differences between participants scoring high versus low on a drive for thinness construct concerning their visual attention toward specific body parts. We hypothesized that participants scoring high on the drive for thinness subscale would show increased attention to body regions, which are important in the assessment of body weight and thinness like the waist, hips, legs, and arms. METHOD: We examined eye-gaze behavior of a nonclinical sample of 51 male and female college students with an eye-tracking system as they were looking at pictures of young, attractive males and females. In addition, we used the Eating Disorder Inventory to measure drive for thinness. RESULTS: Participants with increased scores on the drive for thinness subscale looked longer and more often to the waist, hips, legs, and arms as compared with low scorers. In addition, they showed decreased attention toward the head or face. CONCLUSION: The results indicate that participants scoring high on drive for thinness show an attentional bias toward body regions that are associated with assessing changes in weight. However, they neglected the face, which is the most important source of social and affective information when looking at others.

Attentional Blink to emotional and threatening pictures in spider phobics: electrophysiology and behavior.

When two targets have to be identified in a rapid serial visual presentation (RSVP) paradigm, perception of the second target (T2) becomes significantly impaired if it is displayed 200-500 ms after the first target (T1), a phenomenon labeled as "Attentional Blink" (AB). Here we investigate 14 spider phobics and 16 controls in an RSVP paradigm with neutral T1s. T2 pictures were neutral, emotional (positive or negative) or threatening (spiders for spider phobics). In addition, event-related potentials in response to T2 targets were analyzed. Both spider phobics and controls correctly identified positive and negative T2s more often than neutral T2s, indicating a reduction of the AB effect caused by emotional stimuli. In addition, spider phobics detected spider T2s more frequently than all other T2s. Furthermore, significantly larger P300 amplitudes accompanied detection of spider T2s in the spider phobics as compared to the controls. Based on recent theoretical accounts of the AB effect, results indicate a phobia-related post-perceptual consolidation bias of threatening information in spider phobic subjects.

Decision-making in Blackjack: an electrophysiological analysis.

Previous studies have identified a negative potential in the event-related potential (ERP), the error-related negativity (ERN), which is claimed to be triggered by a deviation from a reward expectation. Furthermore, this negativity is related to shifts in risk taking, strategic behavioral adjustments, and inhibition. We used a computer Blackjack gambling task to further examine the process associated with the ERN. Our findings are in line with the view that the ERN process is related to the degree of reward expectation. Furthermore, increased ERN amplitude is associated with the negative evaluation of ongoing decisions, and the amplitude of the ERN is directly related to risk-taking and decision-making behavior. However, the findings suggest that an explanation exclusively based on the deviation from a reward expectation may be insufficient and that the intention of the participants and the importance of a negative event for learning and behavioral change are crucial to the understanding of ERN phenomena.

Identification, cloning, and functional analysis of the human U6 snRNA-specific terminal uridylyl transferase.

Mammalian cells contain a highly specific terminal uridylyl transferase (TUTase) that exclusively accepts U6 snRNA as substrate. This enzyme, termed U6-TUTase, was purified from HeLa cell extracts and analyzed by microsequencing. All sequenced peptides matched a unique human cDNA coding for a previously unknown protein. Domain structure analysis revealed that the U6-TUTase also belongs to the well-characterized poly(A) polymerase protein superfamily. However, by amino acid sequence as well as RNA-binding motifs, human U6-TUTase is highly divergent from both the poly(A) polymerases and from the TUTases identified within the editing complexes of trypanosomes. After cloning, the recombinant U6-TUTase was expressed in HeLa cells. Analysis of its catalytical activity confirmed the identity of the cloned protein as U6-TUTase, exhibiting the same exclusive substrate specificity for U6 snRNA as the endogenous enzyme. That unique selectivity even excluded as substrate U6atac RNA, the functional homolog of the minor spliceosome. Finally, RNAi knockdown experiments revealed that U6-TUTase is essential for cell proliferation. Surprisingly, large amounts of the recombinant enzyme were found to accumulate within nucleoli.

Electrophysiological correlates of threat processing in spider phobics.

The electrocortical correlates of the processing of feared/fear-relevant and neutral stimuli were investigated in a pictorial emotional Stroop paradigm with spider phobic, social phobic, and nonphobic subjects. Subjects identified either the color of red or blue pictures of spiders, birds, or flowers (emotional Stroop task) or the object itself (identification task) by pressing different buttons. No emotional Stroop interference was found for spider phobic subjects when identifying the color of spiders as opposed to neutral stimuli. However, in the object identification task, spider phobic subjects identified spiders significantly faster than birds or flowers. Parietal P300 and P400 amplitudes were enhanced independent of task in spider phobic but not in nonphobic subjects when viewing pictures of spiders, which is consistent with previous studies showing that highly unpleasant and arousing pictures affect parietal late positive potentials.

Event-related brain potentials and affective responses to threat in spider/snake-phobic and non-phobic subjects.

We investigated cortical responses and valence/arousal ratings of spider phobic, snake phobic, and healthy subjects while they were processing feared, fear-relevant, emotional neutral, and pleasant stimuli. Results revealed significantly larger amplitudes of late ERP components (P3 and late positive complex, LPC) but not of early components (N1, P2, N2) in phobics when subjects were processing feared stimuli. This fear-associated increase of amplitudes of late ERP components in phobic subjects was maximal at centro-parietal and occipital brain sites. Furthermore, phobics but not controls rated feared stimuli to be more negative and arousing than fear-relevant, emotional neutral, and pleasant stimuli. Since late ERP components and valence/arousal ratings were only significantly increased when phobic subjects but not when healthy controls were processing feared stimuli, the present data suggest that P3 and LPC amplitudes represent useful neural correlates of the emotional significance and meaning of stimuli.

Eye movements and behavioral responses to threatening and nonthreatening stimuli during visual search in phobic and nonphobic subjects.

Spider-phobic and nonphobic subjects searched for a feared/fear-relevant (spider) or neutral target (mushroom) presented in visual matrices of neutral objects (flowers). In half of the displays, the mushroom target was paired with a spider distractor, or a spider target was paired with a mushroom distractor. Although all subjects responded faster to the neutral target than to the feared/fear-relevant target, phobics were slower to respond than nonphobics when a mushroom target was presented with a spider distractor. Their eyes appeared to be drawn to the feared distractor before fixating neutral targets. A further experiment indicated no group differences when subjects merely judged the homogeneity of matrices. Thus, threat seems to capture the attention of phobics only when it is part of a background that subjects are explicitly instructed to ignore.

Acupuncture decreases somatosensory evoked potential amplitudes to noxious stimuli in anesthetized volunteers.

UNLABELLED: The effect of acupuncture on pain perception is controversial. Because late amplitudes of somatosensory evoked potentials (SEPs) to noxious stimuli are thought to correlate with the subjective experience of pain intensity, we designed this study to detect changes of these SEPs before and after acupuncture in a double-blinded fashion. Sixteen volunteers were anesthetized by propofol and exposed to painful electric stimuli to the right forefinger. Then, blinded to the research team, the acupuncture group (n = 8) was treated with electric needle acupuncture over 15 min at analgesic points of the leg, whereas the sham group (n = 8) received no treatment. Thereafter, nociceptive stimulation was repeated. SEPs were recorded during each noxious stimulation from the vertex Cz, and latencies and amplitudes of the N150 and P260 components were analyzed by analysis of variance. P260 amplitudes decreased from 4.40 +/- 2.76 microV (mean +/- SD) before treatment to 1.67 +/- 1.21 microV after treatment (P < 0.05), whereas amplitudes of the sham group remained unchanged (2.64 +/- 0.94 microV before versus 2.54 +/- 1.54 microV after treatment). In conclusion, this double-blinded study demonstrated that electric needle acupuncture, as compared with sham treatment, significantly decreased the magnitudes of late SEP amplitudes with electrical noxious stimulation in anesthetized subjects, suggesting a specific analgesic effect of acupuncture. IMPLICATIONS: This double-blinded study demonstrates that electric needle acupuncture, as compared with sham treatment, significantly decreases the magnitudes of late somatosensory evoked potential amplitudes with electrical noxious stimulation in anesthetized subjects, suggesting a specific analgesic effect of acupuncture.

Biochemical characterization of a U6 small nuclear RNA-specific terminal uridylyltransferase.

The HeLa cell terminal uridylyltransferase (TUTase) that specifically modifies the 3'-end of mammalian U6 small nuclear RNA (snRNA) was characterized with respect to ionic dependence and substrate requirements. Optimal enzyme activity was obtained at moderate ionic strength (60 mm KCl) and depended on the presence of 5 mm MgCl2. In vitro synthesized U6 snRNA without a 3'-terminal UMP residue was not accepted as substrate. In contrast, U6 snRNA molecules containing one, two or three 3'-terminal UMP residues were filled up efficiently, generating the 3'-terminal structure with four UMP residues observed in newly transcribed cellular U6 snRNA. In this reaction, the addition of more than one UMP nucleotide depended on higher UTP concentrations. The analysis of internally mutated U6 snRNA revealed that the fill-in reaction by the U6-TUTase was not controlled by opposite-strand nucleotides, excluding an RNA-dependent RNA polymerase mechanism. Furthermore, electrophoretic mobility-shift analyses showed that the U6-TUTase was able to form stable complexes with the U6 snRNA in vitro. On the basis of these findings, a protocol was developed for affinity purification of the enzyme. In agreement with indirect labeling results, PAGE of a largely purified enzyme revealed an apparent molecular mass of 115 kDa for the U6-TUTase.