Oligomeric and fibrillar species of β-amyloid (Aβ42) both impair mitochondrial function in P301L tau transgenic mice

We recently provided evidence for a mitochondrial dysfunction in P301L tau transgenic mice, a strain modeling the tau pathology of Alzheimer's disease (AD) and frontotemporal dementia (FTD). In addition to tau aggregates, the AD brain is further characterized by Aβ peptide-containing plaques. When we addressed the role of Aβ, this indicated a synergistic action of tau and Aβ pathology on the mitochondria. In the present study, we compared the toxicity of different Aβ42 conformations in light of recent studies suggesting that oligomeric rather than fibrillar Aβ might be the actual toxic species. Interestingly, both oligomeric and fibrillar, but not disaggregated (mainly monomeric) Aβ42 caused a decreased mitochondrial membrane potential in cortical brain cells obtained from FTD P301L tau transgenic mice. This was not observed with cerebellar preparations indicating selective vulnerability of cortical neurons. Furthermore, we found reductions in state 3 respiration, the respiratory control ratio, and uncoupled respiration when incubating P301L tau mitochondria either with oligomeric or fibrillar preparations of Aβ42. Finally, we found that aging specifically increased the sensitivity of mitochondria to oligomeric Aβ42 damage indicating that oligomeric and fibrillar Aβ42 are both toxic, but exert different degrees of toxicit

Optical properties of the human round window membrane

Optical techniques are effective tools for diagnostic applications in medicine and are particularly attractive for the noninvasive analysis of biological tissues and fluids in vivo. Noninvasive examinations of substances via a fiber optic probe need to consider the optical properties of biological tissues obstructing the optical path. This applies to the analysis of the human perilymph, which is located behind the round window membrane. The composition of this inner ear liquid is directly correlated to inner ear hearing loss. In this work, experimental methods for studying the optical properties of the human round window membrane ex vivo are presented. For the first time, a comprehensive investigation of this tissue is performed, including optical transmission, forward scattering, and Raman scattering. The results obtained suggest the application of visible wavelengths (>400nm) for investigating the perilymph behind the round window membrane in future. © 2017 Society of Photo-Optical Instrumentation Engineers (SPIE)

From segment to somite: segmentation to epithelialization analyzed within quantitative frameworks

One of the most visually striking patterns in the early developing embryo is somite segmentation. Somites form as repeated, periodic structures in pairs along nearly the entire caudal vertebrate axis. The morphological process involves short- and long-range signals that drive cell rearrangements and cell shaping to create discrete, epithelialized segments. Key to developing novel strategies to prevent somite birth defects that involve axial bone and skeletal muscle development is understanding how the molecular choreography is coordinated across multiple spatial scales and in a repeating temporal manner. Mathematical models have emerged as useful tools to integrate spatiotemporal data and simulate model mechanisms to provide unique insights into somite pattern formation. In this short review, we present two quantitative frameworks that address the morphogenesis from segment to somite and discuss recent data of segmentation and epithelialization

EGFL7 loss correlates with increased VEGF-D expression, upregulating hippocampal adult neurogenesis and improving spatial learning and memory

Correction: Volume: 80 Issue: 8 DOI: 10.1007/s00018-023-04835-3 Article Number: 201 Published: AUG 2023Neural stem cells reside in the subgranular zone, a specialized neurogenic niche of the hippocampus. Throughout adulthood, these cells give rise to neurons in the dentate gyrus, playing an important role in learning and memory. Given that these core cognitive processes are disrupted in numerous disease states, understanding the underlying mechanisms of neural stem cell proliferation in the subgranular zone is of direct practical interest. Here, we report that mature neurons, neural stem cells and neural precursor cells each secrete the neurovascular protein epidermal growth factor-like protein 7 (EGFL7) to shape this hippocampal niche. We further demonstrate that EGFL7 knock-out in a Nestin-CreERT2-based mouse model produces a pronounced upregulation of neurogenesis within the subgranular zone. RNA sequencing identified that the increased expression of the cytokine VEGF-D correlates significantly with the ablation of EGFL7. We substantiate this finding with intraventricular infusion of VEGF-D upregulating neurogenesis in vivo and further show that VEGF-D knock-out produces a downregulation of neurogenesis. Finally, behavioral studies in EGFL7 knock-out mice demonstrate greater maintenance of spatial memory and improved memory consolidation in the hippocampus by modulation of pattern separation. Taken together, our findings demonstrate that both EGFL7 and VEGF-D affect neurogenesis in the adult hippocampus, with the ablation of EGFL7 upregulating neurogenesis, increasing spatial learning and memory, and correlating with increased VEGF-D expression.Peer reviewe

Characterization of the micro-environment of the testis that shapes the phenotype and function of testicular macrophages

Tissue-specific macrophages are important for the activation of innate immune responses and general organ homeostasis. Testicular macrophages (TM) reside in the testicular interstitial space and comprise the largest leukocyte population in the testis and are assumed to play a role in maintaining testicular immune privilege. Numerous studies have indicated that the interstitial fluid (IF) surrounding the TM has immunosuppressive properties, which may influence the TM phenotype. However, the identity of the immunosuppressive molecules present in the IF is poorly characterized. In this thesis it is shown that in the rat, IF shifts the M1 phenotype of granulocyte macrophage-colony stimulating factor induced bone marrow derived macrophages towards the M2 phenotype. M2 macrophages polarized by IF mimic the properties of TM such as increased expression of CD163, high secretion of IL-10 and low secretion of TNF-alpha. In addition, IF-polarized macrophages display immunoregulatory functions by inducing the expansion of immunosuppressive regulatory T cells. This thesis provides evidence that PGE2, PGI2, testosterone and corticosterone are important immunoregulatory molecules in the IF, playing a relevant role in determining the phenotype of TM. Except corticosterone, all of these factors are able to inhibit the NF-kB signaling pathway to suppress the production of pro-inflammatory cytokines and thus maintain an immunosuppressive microenvironment of the testis. Corticosterone was found to be the principal immunosuppressive molecule in the IF. Its receptor, the glucocorticoid receptor, was found to be present in TM immunohistochemically. In addition, TM locally produce small amounts of corticosterone, which suppress the expression of inflammatory genes and render TM refractory to inflammatory stimuli. Taken together, these results suggest that testicular corticosterone shapes the immunosuppressive function and phenotype of TM. This steroid hormone may therefore play also an important role in the establishment and maintenance of the immune privilege of the testis.Gewebsspezifische Makrophagen haben eine wichtige Funktion bei der Aktivierung angeborener Immunantworten und der Organhomeostase. Testikuläre Makrophagen (TM) befinden sich im Interstitium des Hodens und stellen die größte Leukozytenpopulation in der männlichen Gonade dar. Es wird angenommen, dass sie eine wichtige Funktion in der Aufrechterhaltung des Immunprivilegs des Hodens ausüben. Studien haben gezeigt, dass die interstitielle Flüssigkeit (IF), wleche die TM umgibt, immunsuppressive Eigenschaften aufweist, die den Phänotyp der TM beeinflussen könnten. Allerdings konnten immunsuppressive Moleküle in der IF bislang kaum charakterisiert werden. In der vorliegenden Arbeit wird für die Ratte als Modell gezeigt, dass die IF den durch Granulozyten- Makrophagen-Kolonie-stimulierenden Faktor (GM-CSF) induzierten M1 Phänotyp von Makrophagen, die aus dem Knochenmark isoliert wurden, in Richtung des M2 Phänotyps verschieben kann. IF-polarisierte M2-Makrophagen zeigen damit charakteristische Eigenschaften von TM, wie z. Bsp. erhöhte Expression von CD163, hohe Level von sezerniertem IL-10 bei geringer TNF-alpha Sekretion. Darüber hinaus zeigen IF-polarisierte Makrophagen immunoregulatorische Funktionen, indem sie die Expansion von immunsuppressiven regulatorischen T-Zellen induzieren. In dieser Studie werden erstmals auch Ergebnisse vorgestellt, die zeigen, dass PGE2, PGI2, Testosteron und Corticosteron wichtige immunregulatorische Moleküle in der IF darstellen und eine wesentliche Rolle bei der Bestimmung des TM-Phänotyps spielen. Mit Ausnahme von Corticosteron sind die genannten Faktoren in der Lage, den NF-kB-Signalweg zu hemmen, und damit die Produktion von entzündungshemmenden Zytokinen zu unterdrücken. Bei Corticosteron war der NFkB Signalweg bei der Immunsuppression nicht blockiert. Corticosteron konnte als wichtigster immunsuppressiver Faktor in der IF identifiziert werden. Dessen Rezeptor, der Glucocorticoidrezeptor, konnte in TM mittels Immunhistochemie gefunden werden. TM produzieren lokal moderate Mengen an Corticosteron, die die Expression inflammatorischer Gene unterdrücken und TM unempfindlich gegenüber entzündlichen Stimuli machen können. Zusammengenommen zeigen diese Ergebnisse, dass testikuläres Corticosteron maßgeblich für die immunsuppressive Funktion und den spezifischen Phänotyp der TM verantwortlich ist. Damit könnte das Steroidhormon auch eine wichtige Rolle bei der Etablierung und Aufrechterhaltung des Immunprivilegs im Hoden spielen

Morphogenesis by coupled regulatory networks: Reliable control of positional information and proportion regulation

Based on a non-equilibrium mechanism for spatial pattern formation we study how position information can be controlled by locally coupled discrete dynamical networks, similar to gene regulation networks of cells in a developing multicellular organism. As an example we study the developmental problems of domain formation and proportion regulation in the presence of noise, as well as in the presence of cell flow. We find that networks that solve this task exhibit a hierarchical structure of information processing and are of similar complexity as developmental circuits of living cells. Proportion regulation is scalable with system size and leads to sharp, precisely localized boundaries of gene expression domains, even for large numbers of cells. A detailed analysis of noise-induced dynamics, using a mean-field approximation, shows that noise in gene expression states stabilizes (rather than disrupts) the spatial pattern in the presence of cell movements, both for stationary as well as growing systems. Finally, we discuss how this mechanism could be realized in the highly dynamic environment of growing tissues in multi-cellular organisms.Comment: Journal of Theoretical Biology, in pres

Prognostic impact of urokinase-type plasminogen activator system components in clear cell renal cell carcinoma patients without distant metastasis

Background Members of the urokinase-type plasminogen activator (uPA) system including uPA, its receptor uPAR and the plasminogen activator inhibitor 1 (PAI-1) play an important role in tumour invasion and progression in a variety of tumour types. Since the majority of clear cell renal cell carcinoma (ccRCC) shows distant metastasis at time of diagnosis or later, the interplay of uPA, uPAR and PAI-1 might be of importance in this process determining the patients’ outcome. Methods Corresponding pairs of malignant and non-malignant renal tissue specimens were obtained from 112 ccRCC patients without distant metastasis who underwent tumour nephrectomy. Tissue extracts prepared from fresh-frozen tissue samples by detergent extraction were used for the determination of antigen levels of uPA, uPAR and PAI-1 by ELISA. Antigen levels were normalised to protein concentrations and expressed as ng per mg of total protein. Results Antigen levels of uPA, uPAR, and PAI-1 correlated with each other in the malignant tissue specimens (rs=0.51-0.65; all P<0.001). Antigen levels of uPA system components were significantly higher in tissue extracts of non-organ confined tumours (pT3+4) compared to organ-confined tumours (pT1+2; all P<0.05). Significantly elevated levels of uPAR and PAI-1 were also observed in high grade ccRCC. When using median antigen levels as cut-off points, all three uPA system factors were significant predictors for disease-specific survival (DSS) in univariate Cox’s regression analyses. High levels of uPA and uPAR remained independent predictors for DSS with HR=2.86 (95% CI 1.07-7.67, P=0.037) and HR=4.70 (95% CI 1.51-14.6, P=0.008), respectively, in multivariate Cox’s regression analyses. A combination of high antigen levels of uPA and/or uPAR further improved the prediction of DSS in multivariate analysis (HR=14.5, 95% CI 1.88-111.1, P=0.010). Moreover, high uPA and/or uPAR levels defined a patient subgroup of high risk for tumour-related death in ccRCC patients with organ-confined disease (pT1+2) (HR=9.83, 95% CI 1.21-79.6, P=0.032). Conclusions High levels of uPA and uPAR in tumour tissue extracts are associated with a significantly shorter DSS of ccRCC patients without distant metastases

Characterization of ftsZ Mutations that Render Bacillus subtilis Resistant to MinC

Background: Cell division in Bacillus subtilis occurs precisely at midcell. Positional control of cell division is exerted by two mechanisms: nucleoid occlusion, through Noc, which prevents division through nucleoids, and the Min system, where the combined action of the MinC, D and J proteins prevents formation of the FtsZ ring at cell poles or recently completed division sites. Methodology/Principal Findings: We used a genetic screen to identify mutations in ftsZ that confer resistance to the lethal overexpression of the MinC/MinD division inhibitor. The FtsZ mutants were purified and found to polymerize to a similar or lesser extent as wild type FtsZ, and all mutants displayed reduced GTP hydrolysis activity indicative of a reduced polymerization turnover. We found that even though the mutations conferred in vivo resistance to MinC/D, the purified FtsZ mutants did not display strong resistance to MinC in vitro. Conclusions/Significance: Our results show that in B. subtilis, overproduction of MinC can be countered by mutations that alter FtsZ polymerization dynamics. Even though it would be very likely that the FtsZ mutants found depend on other Z-ring stabilizing proteins such as ZapA, FtsA or SepF, we found this not to be the case. This indicates that the cell division process in B. subtilis is extremely robust.

The ReCoDe addiction research consortium:Losing and regaining control over drug intake-Findings and future perspectives

Substance use disorders (SUDs) are seen as a continuum ranging from goal-directed and hedonic drug use to loss of control over drug intake with aversive consequences for mental and physical health and social functioning. The main goals of our interdisciplinary German collaborative research centre on Losing and Regaining Control over Drug Intake (ReCoDe) are (i) to study triggers (drug cues, stressors, drug priming) and modifying factors (age, gender, physical activity, cognitive functions, childhood adversity, social factors, such as loneliness and social contact/interaction) that longitudinally modulate the trajectories of losing and regaining control over drug consumption under real-life conditions. (ii) To study underlying behavioural, cognitive and neurobiological mechanisms of disease trajectories and drug-related behaviours and (iii) to provide non-invasive mechanism-based interventions. These goals are achieved by: (A) using innovative mHealth (mobile health) tools to longitudinally monitor the effects of triggers and modifying factors on drug consumption patterns in real life in a cohort of 900 patients with alcohol use disorder. This approach will be complemented by animal models of addiction with 24/7 automated behavioural monitoring across an entire disease trajectory; i.e. from a naïve state to a drug-taking state to an addiction or resilience-like state. (B) The identification and, if applicable, computational modelling of key molecular, neurobiological and psychological mechanisms (e.g., reduced cognitive flexibility) mediating the effects of such triggers and modifying factors on disease trajectories. (C) Developing and testing non-invasive interventions (e.g., Just-In-Time-Adaptive-Interventions (JITAIs), various non-invasive brain stimulations (NIBS), individualized physical activity) that specifically target the underlying mechanisms for regaining control over drug intake. Here, we will report on the most important results of the first funding period and outline our future research strategy.</p

Standards in semen examination:publishing reproducible and reliable data based on high-quality methodology

Biomedical science is rapidly developing in terms of more transparency, openness and reproducibility of scientific publications. This is even more important for all studies that are based on results from basic semen examination. Recently two concordant documents have been published: the 6th edition of the WHO Laboratory Manual for the Examination and Processing of Human Semen, and the International Standard ISO 23162:2021. With these tools, we propose that authors should be instructed to follow these laboratory methods in order to publish studies in peer-reviewed journals, preferable by using a checklist as suggested in an Appendix to this article.Peer reviewe