Loss of Bmi1 causes anomalies in retinal development and degeneration of cone photoreceptors.

Retinal development occurs through the sequential but overlapping generation of six types of neuronal cells and one glial cell type. Of these, rod and cone photoreceptors represent the functional unit of light detection and phototransduction and are frequently affected in retinal degenerative diseases. During mouse development, the Polycomb group protein Bmi1 is expressed in immature retinal progenitors and differentiated retinal neurons, including cones. We show here that Bmi1 is required to prevent post natal degeneration of cone photoreceptors and bipolar neurons and that inactivation of Chk2 or p53 could improve but not overcome cone degeneration in Bmi1(-/-) mice. The retinal phenotype of Bmi1(-/-) mice was also characterized by loss of heterochromatin, activation of tandem repeats, oxidative stress and Rip3-associated necroptosis. In the human retina, BMI1 was preferentially expressed in cones at heterochromatic foci. BMI1 inactivation in human embryonic stem cells was compatible with retinal induction but impaired cone terminal differentiation. Despite this developmental arrest, BMI1-deficient cones recapitulated several anomalies observed in Bmi1(-/-) photoreceptors, such as loss of heterochromatin, activation of tandem repeats and induction of p53, revealing partly conserved biological functions between mouse and man.

Sex-specific involvement of the Notch-JAG pathway in social recognition.

Under the hypothesis that olfactory neural epithelium gene expression profiles may be useful to look for disease-relevant neuronal signatures, we examined microarray gene expression in olfactory neuronal cells and underscored Notch-JAG pathway molecules in association with schizophrenia (SZ). The microarray profiling study underscored JAG1 as the most promising candidate. Combined with further validation with real-time PCR, downregulation of NOTCH1 was statistically significant. Accordingly, we reverse-translated the significant finding from a surrogate tissue for neurons, and studied the behavioral profile of Notch1+/- mice. We found a specific impairment in social novelty recognition, whereas other behaviors, such as sociability, novel object recognition and olfaction of social odors, were normal. This social novelty recognition deficit was male-specific and was rescued by rapamycin treatment. Based on the results from the animal model, we next tested whether patients with psychosis might have male-specific alterations in social cognition in association with the expression of NOTCH1 or JAG1. In our first episode psychosis cohort, we observed a specific correlation between the expression of JAG1 and a face processing measure only in male patients. The expression of JAG1 was not correlated with any other cognitive and symptomatic scales in all subjects. Together, although we acknowledge the pioneering and exploratory nature, the present work that combines both human and animal studies in a reciprocal manner suggests a novel role for the Notch-JAG pathway in a behavioral dimension(s) related to social cognition in psychotic disorders in a male-specific manner.

Development of a GMP Phase III purification process for VB4-845, an immunotoxin expressed in E. coli using high cell density fermentation.

VB4-845 is a recombinant immunotoxin comprised of an anti-epithelial cell adhesion molecule (EpCAM) scFv fused to a truncated form of the bacterial toxin, Pseudomonas exotoxin A. VB4-845, purified from TB fed-batch fermentation, showed clinical efficacy when administered locally to treat non-muscle invasive bladder cancer (NMIBC) and squamous cell carcinomas of the head and neck (SCCHN). Here, we describe the implementation of an Escherichia coli high cell density (HCD) cultivation and purification process for VB4-845. HCD cultivation was a prerequisite for achieving higher yields necessary for Phase III clinical trials and commercialization. Using this process, the VB4-845 titer in the supernatant was increased by 30-fold over the original TB fed-batch cultivation. To obtain clinical grade material, a process involving a five-step column purification procedure was implemented and led to an overall recovery of ∼ 40%. VB4-845 purity of >97% was achieved after the first three columns following the removal of low-molecular weight product-related impurities and aggregates. Endotoxins were effectively separated from VB4-845 on the Q-columns and by washing the Ni-column with a detergent buffer while host cell proteins were removed using ceramic hydroxyapatite. Comparability studies demonstrated that the purified product from the Phase III process was identical to the Phase II reference standard produced using TB fed-batch fermentation.

Timely use of in-car dim blue light and blue blockers in the morning does not improve circadian adaptation of fast rotating shift workers.

Circadian adaptation to night work usually does not occur in naturalistic conditions, largely due to exposure to low levels of light during the night and light in the morning on the way home. This leads to circadian misalignment, which has documented deleterious effects on sleep and functioning during waking hours. Chronic circadian misalignment is also being increasingly associated with long-term health comorbidities. As the circadian system is mostly sensitive to short wavelengths (i.e., blue light) and less sensitive to long wavelengths (i.e., red light), shaping light exposure in a "wavelength-wise" manner has been proposed to promote partial adaptation to night shifts, and, therefore, alleviate circadian rhythms disruption. This report presents results from two cross-over designed studies that aimed to investigate the effects of three different light conditions on circadian phase, sleepiness, and alertness of police patrol officers on a rotating shift schedule. The first study took place during summer (n = 15) and the second study (n = 25) during winter/early spring. In both studies, all participants went through three conditions composed of four consecutive night shifts: 1) in-car dim blue light exposure during the night shift and wearing of blue-blocking glasses (BBG) in the morning after 05:00 h; 2) in-car red light exposure during the night shift and wearing of BBG in the morning after 05:00 h; 3) a control condition with no intervention. To assess circadian phase position, salivary melatonin was collected hourly the night before and the night after each condition. Sleep was monitored by wrist actigraphy. Also, a 10-min Psychomotor Vigilance-Task was administered at the beginning and end of each night shift and the Karolinska Sleepiness Scale was completed every 2 h during each night shift. In the summer study, no difference was found in alertness and sleepiness between conditions. Participants though exhibited greater (≈3 h) phase delay after four consecutive night shifts in the control condition (in which morning light exposure was expected to prevent phase delay) than after the blue and red conditions (≈2 h) (in which wearing BBG were expected to promote phase delay). In the second study performed during the winter/early spring, a comparable ≈2 h phase delay was found in each of the three conditions, with no difference in alertness and sleepiness between conditions. In conclusion, participants in both studies exhibited modest phase delay across the four night shifts, even during the control conditions. Still, re-entrainment was not fast enough to produce partial circadian adaptation after four night shifts. A greater number of consecutive night shifts may be necessary to produce enough circadian alignment to elicit benefits on sleepiness and alertness in workers driving a motorized vehicle during night shifts. In-car dim blue light exposure combined with the wearing of BBG in the morning did not show the expected benefits on circadian adaptation, sleepiness, and alertness in our studies. Higher levels of light may be warranted when implementing light intervention in a motorized vehicle setting.

The Electroretinogram May Differentiate Schizophrenia From Bipolar Disorder.

BACKGROUND: The retina is recognized as an approachable part of the brain owing to their common embryonic origin. The electroretinogram (ERG) has proved to be a valuable tool to investigate psychiatric disorders. We therefore investigated its accuracy as a tool to differentiate schizophrenia (SZ) from bipolar disorder (BP) even after balancing patients for their main antipsychotic medication. METHODS: ERG cone and rod luminance response functions were recorded in 150 patients with SZ and 151 patients with BP and compared with 200 control subjects. We created a subgroup of subjects-45 with SZ and 45 with BP-balanced for their main antipsychotic medication. RESULTS: A reduced cone a-wave amplitude and a prolonged b-wave latency were observed in both disorders, whereas a reduced cone b-wave amplitude was present in SZ only. Reduced mixed rod-cone a- and b-wave amplitudes were observed in both disorders. Patients with SZ were distinguishable from control subjects with 0.91 accuracy, 77% sensitivity, and 91% specificity with similar numbers for patients with BP (0.89, 76%, and 88%, respectively). Patients with SZ and patients with BP could be differentiated with an accuracy of 0.86 (whole sample) and 0.83 (subsamples of 45 patients with 80% sensitivity and 82% specificity). Antipsychotic dosages were not correlated with ERG parameters. CONCLUSIONS: The ERG waveform parameters used in this study provided a very accurate distinction between the two disorders when using a logistic regression model. This supports the ERG as a tool that could aid the clinician in the differential diagnosis of SZ and BP in stabilized medicated patients.

A methodology for discovering novel brain-relevant peptides: Combination of ribosome profiling and peptidomics.

Brain derived peptides function as signaling molecules in the brain and regulate various physiological and behavioral processes. The low abundance and atypical fragmentation of these brain derived peptides makes detection using traditional proteomic methods challenging. In this study, we introduce and validate a new methodology for the discovery of novel peptides derived from mammalian brain. This methodology combines ribosome profiling and mass spectrometry-based peptidomics. Using this framework, we have identified a novel peptide in mouse whole brain whose expression is highest in the basal ganglia, hypothalamus and amygdala. Although its functional role is unknown, it has been previously detected in peripheral tissue as a component of the mRNA decapping complex. Continued discovery and studies of novel regulating peptides in mammalian brain may also provide insight into brain disorders.

Application of olfactory tissue and its neural progenitors to schizophrenia and psychiatric research.

PURPOSE OF REVIEW: The goal of this review article is to introduce olfactory epithelium-derived cell/tissue models as a promising surrogate system to study the molecular mechanisms implicated in schizophrenia and other neuropsychiatric disorders. Here, we particularly focus on the utility of their neural progenitors. RECENT FINDINGS: Recent investigations of the pathophysiology of schizophrenia using olfactory epithelium-derived tissue/cell models have provided insights about schizophrenia-associated alterations in neurodevelopment, stress response, and gene/protein expression regulatory pathways. SUMMARY: The olfactory epithelium retains the capacity for lifelong neurogenesis and regeneration, because of the presence of neural stem cells and progenitors. Thus, both mature neurons and neural progenitors can be obtained from the olfactory epithelium without the need for genetic reprogramming and related confounds. Furthermore, the olfactory epithelium is highly scalable resource in translational settings. Here, we also demonstrate recent findings from research using olfactory epithelium-derived tissue/cell models in schizophrenia and other brain disorders. In summary, we propose that the olfactory epithelium is a promising resource to study neural molecular and cellular signatures relevant to the pathology of schizophrenia and other mental disorders.

The Olfactory Neural Epithelium As a Tool in Neuroscience.

Capturing both dynamic changes (state) and persistent signatures (trait) directly associated with disease at the molecular level is crucial in modern medicine. The olfactory neural epithelium, easily accessible in clinical settings, is a promising surrogate model in translational brain medicine, complementing the limitations in current engineered cell models.

The emerging field of retinal electrophysiological measurements in psychiatric research: A review of the findings and the perspectives in major depressive disorder.

Major depressive disorder (MDD) is a severe mental illness leading to long-term disabilities. One of the current challenges in psychiatric research is to develop new approaches to investigate the pathophysiology of MDD and monitor drug response in order to provide better therapeutic strategies to the patients. Since the retina is considered as part of the central nervous system, it was suggested that it constitutes an appropriate site to investigate mental illnesses. In the past years, several teams assessed the retinal function of patients with mood disorders and many relevant abnormalities have been reported. Investigation of the retinal electrophysiological abnormalities in MDD remains a young emerging field, but we believe that the current findings are very promising and we argue that objective retinal electrophysiological measurements may eventually become relevant tools to investigate the pathophysiology of MDD. Here, we review the retinal abnormalities detected with objective electrophysiological measurements such as the flash electroretinogram (fERG), the pattern electroretinogram (PERG) and the electrooculogram (EOG) in patients with MDD. We discuss how these changes might reflect the pathophysiology of MDD in both clinical and scientific points of view, according especially to the monoamine neurotransmission deficiency hypothesis. We also discuss the technical details that must be taken into consideration for a potential use of the objective retinal electrophysiological measurements as tools to investigate the pathophysiology of MDD.

The brain through the retina: the flash electroretinogram as a tool to investigate psychiatric disorders.

Investigating the living brain remains one of the major obstacles in psychiatry research in order to better understand the biological underpinning of brain disorders. Novel approaches are needed to study brain functions indirectly. Since it is part of the central nervous system, retinal functions as measured with the flash electroretinogram (ERG) may reflect the central dysfunctions reported in psychiatric disorders. This review describes the flash ERG anomalies reported in patients with psychiatric disorders such as seasonal affective disorder, schizophrenia, autism spectrum disorder and drug addiction and discusses how changes in retinal functions might be used as biomarkers for psychiatric disorder as well as a potential aid to diagnosis in psychiatry.

Glycogen synthase kinase-3 overexpression replicates electroretinogram anomalies of offspring at high genetic risk for schizophrenia and bipolar disorder.

BACKGROUND: Electroretinogram (ERG) anomalies occur in patients with psychiatric disorders and represent potential biomarkers for diagnosis. For instance, decreased rod ERG (b-wave amplitude at Vmax) is a biological endophenotype in young offspring at high genetic risk (HR) for schizophrenia (SZ) and bipolar disorder (BD). Also, a decrease in cone a-wave and rod a- and b- wave was observed in SZ patients. However, the biological underpinning of these anomalies remains unknown. Several genetic variants associated with enhanced risk for SZ and/or BD can activate glycogen synthase kinase-3 isozymes (GSK3α and ß). Here we examined the potential contribution of GSK3α and ß in the modulation of the ERG. METHODS: Cone and rod ERGs were recorded in mice having increased (prpGSK3ß mice) or reduced (GSK3ß(+/-) mice) GSK3ß expression and in GSK3α knockout (KO) mice. RESULTS: In prpGSK3ß mice, we observed a decrease in rod b-wave amplitude at Vmax, whereas enhanced b-wave amplitude at Vmax was found in GSK3ß(+/-) mice. An increase in cone a- and b-wave amplitude at Vmax and in rod b-wave amplitude at Vmax was observed in GSK3α-KO mice. CONCLUSIONS: GSK3 expression modulates some ERG parameters. The phenotype observed in prpGSK3ß mice is consistent with observations made in HRs. ERG anomalies observed in GSK3ß(+/-) and GSK3α-KO mice confirm an association between the rod and cone b-wave amplitude and the expression of GSK3 isozymes. Changes in GSK3 expression or activity may explain some ERG anomalies in HRs and patients, thus supporting the biological validity of ERG measurements as a valuable biomarker for psychiatric research.

The electroretinogram as a biomarker of central dopamine and serotonin: potential relevance to psychiatric disorders.

BACKGROUND: Dysfunctions in brain dopamine and serotonin neurotransmission are believed to be involved in the etiology of psychiatric disorders, and electroretinogram (ERG) anomalies have been reported in psychiatric patients. The goal of this study was to evaluate whether ERG anomalies could result from central dopamine or serotonin dysfunctions or from changes in the retinal bioavailability of these neurotransmitters. METHOD: Photopic and scotopic ERGs were recorded in R439H tryptophan hydroxylase 2 knockin (Tph2-KI) mice that have an approximately 80% decrease in brain serotonin and dopamine transporter knockout (DAT-KO) mice showing a fivefold increase in brain extracellular dopamine. Dopamine and serotonin retinal and striatal tissue content were also measured. The role of dopamine D1 receptors (D1R) and D2 receptors (D2R) in the ERG responses was evaluated in D1R-KO and D2R-KO mice. RESULTS: An increase in photopic b-wave implicit time was observed in Tph2-KI mice (wildtype = 24.25 msec, KI = 25.22 msec; p = .011). The DAT-KO mice showed a decrease in rod sensitivity (wildtype =-1.97 log units, KO =-1.81 log units; p = .014). In contrast to remarkable alterations in brain levels, no changes in dopamine and serotonin retinal content were found in DAT-KO and Tph2-KI mice, respectively. The D1R-KO mice showed anomalies in photopic and scotopic maximal amplitude, whereas D2R-KO mice showed higher oscillatory potentials relative contribution to the b-wave amplitude. CONCLUSION: Alterations in central dopamine and serotonin neurotransmission can affect the ERG responses. The ERG anomalies reported in psychiatric disorders might serve as biomarkers of central monoaminergic dysfunction, thus promoting ERG measurements as a useful tool in psychiatric research.

Glycogen synthase kinase-3ß haploinsufficiency lengthens the circadian locomotor activity period in mice.

The mood stabiliser drug lithium has been reported to impact circadian rhythms in vertebrates. Among several putative therapeutic molecular targets, direct inhibition of glycogen synthase kinase-3 beta (GSK3ß) by lithium has been proposed to underlie its effects on circadian physiology. Here we study the effect of GSK3ß haploinsufficiency on the circadian locomotor activity in mice during a free-running period in comparison to wildtype littermates (WT). Mice were housed individually to record their circadian wheel running activity and were entrained to a 12h light/12h dark cycle for 14 days and then placed under constant darkness for 14 days to allow free-running. During the free-running phase, the circadian locomotor activity period of GSK3ß(+/-) was significantly lengthened (23.83±0.05h) when compared to the WT mice (23.54±0.10h; p=0.0374). No significant difference in locomotor activity was observed. Knowing that GSK3ß interacts with most of the core clock components, these data suggest that GSK3ß acts as a critical intrinsic regulator of the circadian clock and plays an important role in regulating its period in response to lithium treatment.

Negative impact of melatonin ingestion on the photopic electroretinogram of dogs.

Melatonin follows a circadian rhythm entrained by the light/dark cycle and plays a role in promoting light sensitivity at night. It has been suggested that melatonin and dopamine reciprocal inhibition may contribute to the switch between day and night vision. The purpose of this study was to investigate the impact of a high dose of melatonin administration on the photopic and scotopic electroretinogram (ERG) of dogs in the daytime, when it is not thought to be present. Photopic and scotopic ERG luminance response functions were obtained from 7 anaesthetized beagle dogs (3 males and 4 females), once without melatonin (control) and once after oral administration of melatonin (90 mg/dog). Vmax (maximal b-wave amplitude achieved) and logK (retinal sensitivity) were calculated from the derived luminance response function. Photopic flicker ERG was also recorded. In photopic condition, a-wave amplitude (control: -126.90 µV; with melatonin: -49.64 µV; p<0.001) and Vmax (control: 252.50 µV; with melatonin: 115.40 µV; p<0.001) were decreased. A significant reduction of the photopic flicker ERG amplitude was observed after melatonin ingestion. In scotopic condition, an overall difference was reported before and after melatonin ingestion for the a- and b-wave amplitude, but no change was significant for Vmax. Melatonin ingestion at a high dose during the day decreases the photopic amplitude of a- and b-wave, but has no impact on implicit time. This negative impact of melatonin on photopic system may serve to promote night vision.