Targeting microglial CSF1R in health and disease

The main effector functions of microglia are immune, synaptic network refinement, brain insult reaction, and neurogenesis regulation. This cumulative doctoral thesis focuses on the relevance of the colony-stimulating factor 1 receptor (CSF1R) signaling pathway of microglia in the context of the last two topics, specifically in [A] low-grade white matter inflammation and [B] hypoxia-induced neurogenesis. Ad [A]: Our group previously detected that myelin structural protein cyclic nucleotide phosphodiesterase (CNP) deletion triggers low-grade white matter inflammation and causes a behavioral phenotype named catatonia. Treatment with the CSF1R inhibitor PLX5622 of Cnp-/- mice leads to a strong reduction of neuroinflammation, i.e. microglial numbers and activation status, as well as a clear improvement of the catatonic signs. In my first paper, I systematically addressed aspects which are important for clinical translation. Amongst others, I found that microglia surviving PLX5622-induced depletion display a pro-inflammatory phenotype including targeted phagocytosis of oligodendrocyte precursor cells, and that two PLX5622 treatment cycles are not superior to one. These results may be helpful for guiding future use of CSF1R inhibitors in (pre)clinical studies. Ad [B]: As work of our group has shown, exposure of mice to hypoxia, whether inspiratory or functional by motor-cognitive challenge, triggers the expression of brain erythropoietin (EPO). In my second paper, I found that treatment with exogenous EPO leads to not only increased hippocampal cornu ammonis 1 (CA1) pyramidal neuron numbers and dendritic spine densities, but most importantly it simultaneously decreases microglia proliferation, activity, and motility. Searching for mechanisms, I discovered a direct effect of EPO on microglia that acted in two phases: first EPO triggered immediate microglia apoptosis for just a limited time, leading to decreased microglia numbers. Subsequently, the reduction of the microglia population was maintained by decreased microglia proliferation. This was likely due to an EPO-induced decrease in the expression of interleukin 34, a neuronally expressed ligand of CSF1R. EPO also led to decreased microglia-neuron contacts and microglial metabolism in the CA1. Furthermore, this was paralleled by an increase in intermediate neuronal progenitors, which became mature by the end of a 3-week EPO treatment. Importantly, these effects are dependent on EPO receptor expression in microglia and neurons.2021-09-0

Mitochondrial physiology

As the knowledge base and importance of mitochondrial physiology to evolution, health and disease expands, the necessity for harmonizing the terminology concerning mitochondrial respiratory states and rates has become increasingly apparent. The chemiosmotic theory establishes the mechanism of energy transformation and coupling in oxidative phosphorylation. The unifying concept of the protonmotive force provides the framework for developing a consistent theoretical foundation of mitochondrial physiology and bioenergetics. We follow the latest SI guidelines and those of the International Union of Pure and Applied Chemistry (IUPAC) on terminology in physical chemistry, extended by considerations of open systems and thermodynamics of irreversible processes. The concept-driven constructive terminology incorporates the meaning of each quantity and aligns concepts and symbols with the nomenclature of classical bioenergetics. We endeavour to provide a balanced view of mitochondrial respiratory control and a critical discussion on reporting data of mitochondrial respiration in terms of metabolic flows and fluxes. Uniform standards for evaluation of respiratory states and rates will ultimately contribute to reproducibility between laboratories and thus support the development of data repositories of mitochondrial respiratory function in species, tissues, and cells. Clarity of concept and consistency of nomenclature facilitate effective transdisciplinary communication, education, and ultimately further discovery

Mitochondrial physiology

As the knowledge base and importance of mitochondrial physiology to evolution, health and disease expands, the necessity for harmonizing the terminology concerning mitochondrial respiratory states and rates has become increasingly apparent. The chemiosmotic theory establishes the mechanism of energy transformation and coupling in oxidative phosphorylation. The unifying concept of the protonmotive force provides the framework for developing a consistent theoretical foundation of mitochondrial physiology and bioenergetics. We follow the latest SI guidelines and those of the International Union of Pure and Applied Chemistry (IUPAC) on terminology in physical chemistry, extended by considerations of open systems and thermodynamics of irreversible processes. The concept-driven constructive terminology incorporates the meaning of each quantity and aligns concepts and symbols with the nomenclature of classical bioenergetics. We endeavour to provide a balanced view of mitochondrial respiratory control and a critical discussion on reporting data of mitochondrial respiration in terms of metabolic flows and fluxes. Uniform standards for evaluation of respiratory states and rates will ultimately contribute to reproducibility between laboratories and thus support the development of data repositories of mitochondrial respiratory function in species, tissues, and cells. Clarity of concept and consistency of nomenclature facilitate effective transdisciplinary communication, education, and ultimately further discovery

CaMKIIα Expressing Neurons to Report Activity-Related Endogenous Hypoxia upon Motor-Cognitive Challenge

We previously introduced the brain erythropoietin (EPO) circle as a model to explain the adaptive ‘brain hardware upgrade’ and enhanced performance. In this fundamental circle, brain cells, challenged by motor-cognitive tasks, experience functional hypoxia, triggering the expression of EPO among other genes. We attested hypoxic cells by a transgenic reporter approach under the ubiquitous CAG promoter, with Hif-1α oxygen-dependent degradation-domain (ODD) fused to CreERT2-recombinase. To specifically focus on the functional hypoxia of excitatory pyramidal neurons, here, we generated CaMKIIα-CreERT2-ODD::R26R-tdTomato mice. Behavioral challenges, light-sheet microscopy, immunohistochemistry, single-cell mRNA-seq, and neuronal cultures under normoxia or hypoxia served to portray these mice. Upon complex running wheel performance as the motor-cognitive task, a distinct increase in functional hypoxic neurons was assessed immunohistochemically and confirmed three-dimensionally. In contrast, fear conditioning as hippocampal stimulus was likely too short-lived to provoke neuronal hypoxia. Transcriptome data of hippocampus under normoxia versus inspiratory hypoxia revealed increases in CA1 CaMKIIα-neurons with an immature signature, characterized by the expression of Dcx, Tbr1, CaMKIIα, Tle4, and Zbtb20, and consistent with accelerated differentiation. The hypoxia reporter response was reproduced in vitro upon neuronal maturation. To conclude, task-associated activity triggers neuronal functional hypoxia as a local and brain-wide reaction mediating adaptive neuroplasticity. Hypoxia-induced genes such as EPO drive neuronal differentiation, brain maturation, and improved performance

Erythropoietin re-wires cognition-associated transcriptional networks

Abstract Recombinant human erythropoietin (rhEPO) has potent procognitive effects, likely hematopoiesis-independent, but underlying mechanisms and physiological role of brain-expressed EPO remained obscure. Here, we provide transcriptional hippocampal profiling of male mice treated with rhEPO. Based on ~108,000 single nuclei, we unmask multiple pyramidal lineages with their comprehensive molecular signatures. By temporal profiling and gene regulatory analysis, we build developmental trajectory of CA1 pyramidal neurons derived from multiple predecessor lineages and elucidate gene regulatory networks underlying their fate determination. With EPO as ‘tool’, we discover populations of newly differentiating pyramidal neurons, overpopulating to ~200% upon rhEPO with upregulation of genes crucial for neurodifferentiation, dendrite growth, synaptogenesis, memory formation, and cognition. Using a Cre-based approach to visually distinguish pre-existing from newly formed pyramidal neurons for patch-clamp recordings, we learn that rhEPO treatment differentially affects excitatory and inhibitory inputs. Our findings provide mechanistic insight into how EPO modulates neuronal functions and networks

Hippocampal neurons respond to brain activity with functional hypoxia

Physical activity and cognitive challenge are established non-invasive methods to induce comprehensive brain activation and thereby improve global brain function including mood and emotional well-being in healthy subjects and in patients. However, the mechanisms underlying this experimental and clinical observation and broadly exploited therapeutic tool are still widely obscure. Here we show in the behaving brain that physiological (endogenous) hypoxia is likely a respective lead mechanism, regulating hippocampal plasticity via adaptive gene expression. A refined transgenic approach in mice, utilizing the oxygen-dependent degradation (ODD) domain of HIF-1α fused to CreERT2 recombinase, allows us to demonstrate hypoxic cells in the performing brain under normoxia and motor-cognitive challenge, and spatially map them by light-sheet microscopy, all in comparison to inspiratory hypoxia as strong positive control. We report that a complex motor-cognitive challenge causes hypoxia across essentially all brain areas, with hypoxic neurons particularly abundant in the hippocampus. These data suggest an intriguing model of neuroplasticity, in which a specific task-associated neuronal activity triggers mild hypoxia as a local neuron-specific as well as a brain-wide response, comprising indirectly activated neurons and non-neuronal cells

BIN1K358R suppresses glial response to plaques in mouse model of Alzheimer's disease

IntroductionThe BIN1 coding variant rs138047593 (K358R) is linked to Late-Onset Alzheimer's Disease (LOAD) via targeted exome sequencing.MethodsTo elucidate the functional consequences of this rare coding variant on brain amyloidosis and neuroinflammation, we generated BIN1K358R knock-in mice using CRISPR/Cas9 technology. These mice were subsequently bred with 5xFAD transgenic mice, which serve as a model for Alzheimer's pathology.ResultsThe presence of the BIN1K358R variant leads to increased cerebral amyloid deposition, with a dampened response of astrocytes and oligodendrocytes, but not microglia, at both the cellular and transcriptional levels. This correlates with decreased neurofilament light chain in both plasma and brain tissue. Synaptic densities are significantly increased in both wild-type and 5xFAD backgrounds homozygous for the BIN1K358R variant.DiscussionThe BIN1 K358R variant modulates amyloid pathology in 5xFAD mice, attenuates the astrocytic and oligodendrocytic responses to amyloid plaques, decreases damage markers, and elevates synaptic densities.HighlightsBIN1 rs138047593 (K358R) coding variant is associated with increased risk of LOAD. BIN1 K358R variant increases amyloid plaque load in 12-month-old 5xFAD mice. BIN1 K358R variant dampens astrocytic and oligodendrocytic response to plaques. BIN1 K358R variant decreases neuronal damage in 5xFAD mice. BIN1 K358R upregulates synaptic densities and modulates synaptic transmission

Polarization Properties of the Weakly Magnetized Neutron Star X-Ray Binary GS 1826–238 in the High Soft State

The launch of the Imaging X-ray Polarimetry Explorer (IXPE) on 2021 December 9 has opened a new window in X-ray astronomy. We report here the results of the first IXPE observation of a weakly magnetized neutron star, GS 1826−238, performed on 2022 March 29–31 when the source was in a high soft state. An upper limit (99.73% confidence level) of 1.3% for the linear polarization degree is obtained over the IXPE 2–8 keV energy range. Coordinated INTEGRAL and NICER observations were carried out simultaneously with IXPE. The spectral parameters obtained from the fits to the broadband spectrum were used as inputs for Monte Carlo simulations considering different possible geometries of the X-ray emitting region. Comparing the IXPE upper limit with these simulations, we can put constraints on the geometry and inclination angle of GS 1826–238