The transcription factor Aiolos restrains the activation of intestinal intraepithelial lymphocytes.

Intestinal intraepithelial lymphocytes (IELs) exhibit prompt innate-like responses to microenvironmental cues and require strict control of effector functions. Here we showed that Aiolos, an Ikaros zinc-finger family member encoded by Ikzf3, acted as a regulator of IEL activation. Ikzf3-/- CD8αα+ IELs had elevated expression of NK receptors, cytotoxic enzymes, cytokines and chemokines. Single-cell RNA sequencing of Ikzf3-/- and Ikzf3+/+ IELs showed an amplified effector machinery in Ikzf3-/- CD8αα+ IELs compared to Ikzf3+/+ counterparts. Ikzf3-/- CD8αα+ IELs had increased responsiveness to interleukin-15, which explained a substantial part, but not all, of the observed phenotypes. Aiolos binding sites were close to those for the transcription factors STAT5 and RUNX, which promote interleukin-15 signaling and cytolytic programs, and Ikzf3 deficiency partially increased chromatin accessibility and histone acetylation in these regions. Ikzf3 deficiency in mice enhanced susceptibility to colitis, underscoring the relevance of Aiolos in regulating the effector function in IELs.

Dntt expression reveals developmental hierarchy and lineage specification of hematopoietic progenitors.

Intrinsic and extrinsic cues determine developmental trajectories of hematopoietic stem cells (HSCs) towards erythroid, myeloid and lymphoid lineages. Using two newly generated transgenic mice that report and trace the expression of terminal deoxynucleotidyl transferase (TdT), transient induction of TdT was detected on a newly identified multipotent progenitor (MPP) subset that lacked self-renewal capacity but maintained multilineage differentiation potential. TdT induction on MPPs reflected a transcriptionally dynamic but uncommitted stage, characterized by low expression of lineage-associated genes. Single-cell CITE-seq indicated that multipotency in the TdT+ MPPs is associated with expression of the endothelial cell adhesion molecule ESAM. Stable and progressive upregulation of TdT defined the lymphoid developmental trajectory. Collectively, we here identify a new multipotent progenitor within the MPP4 compartment. Specification and commitment are defined by downregulation of ESAM which marks the progressive loss of alternative fates along all lineages.

Progenitors of distinct lineages shape the diversity of mature type 2 conventional dendritic cells.

Conventional dendritic cells (cDC) are antigen-presenting cells comprising cDC1 and cDC2, responsible for priming naive CD8+ and CD4+ T cells, respectively. Recent studies have unveiled cDC2 heterogeneity and identified various cDC2 progenitors beyond the common DC progenitor (CDP), hinting at distinct cDC2 lineages. By generating Cd300ciCre-hCD2R26tdTomato reporter mice, we identified a bone marrow pro-cDC2 progenitor exclusively generating cDC2 in vitro and in vivo. Single-cell analyses and multiparametric flow cytometry demonstrated that pro-cDC2 encompasses myeloid-derived pre-cDC2 and lymphoid-derived plasmacytoid DC (pDC)-like precursors differentiating into a transcriptionally convergent cDC2 phenotype. Cd300c-traced cDC2 had distinct transcriptomic profiles, phenotypes, and tissue distributions compared with Ms4a3CreR26tdTomato lineage-traced DC3, a monocyte-DC progenitor (MDP)-derived subset that bypasses CDP. Mice with reduced Cd300c-traced cDC2 showed impaired humoral responses to T cell-dependent antigens. We conclude that progenitors of distinct lineages shape the diversity of mature cDC2 across tissues. Thus, ontogenesis may impact tissue immune responses.

A Cre-deleter specific for embryo-derived brain macrophages reveals distinct features of microglia and border macrophages.

Genetic tools to target microglia specifically and efficiently from the early stages of embryonic development are lacking. We generated a constitutive Cre line controlled by the microglia signature gene Crybb1 that produced nearly complete recombination in embryonic brain macrophages (microglia and border-associated macrophages [BAMs]) by the perinatal period, with limited recombination in peripheral myeloid cells. Using this tool in combination with Flt3-Cre lineage tracer, single-cell RNA-sequencing analysis, and confocal imaging, we resolved embryonic-derived versus monocyte-derived BAMs in the mouse cortex. Deletion of the transcription factor SMAD4 in microglia and embryonic-derived BAMs using Crybb1-Cre caused a developmental arrest of microglia, which instead acquired a BAM specification signature. By contrast, the development of genuine BAMs remained unaffected. Our results reveal that SMAD4 drives a transcriptional and epigenetic program that is indispensable for the commitment of brain macrophages to the microglia fate and highlight Crybb1-Cre as a tool for targeting embryonic brain macrophages.

Distinct progenitor lineages contribute to the heterogeneity of plasmacytoid dendritic cells.

Plasmacytoid dendritic cells (pDCs) are an immune subset devoted to the production of high amounts of type 1 interferons in response to viral infections. Whereas conventional dendritic cells (cDCs) originate mostly from a common dendritic cell progenitor (CDP), pDCs have been shown to develop from both CDPs and common lymphoid progenitors. Here, we found that pDCs developed predominantly from IL-7R+ lymphoid progenitor cells. Expression of SiglecH and Ly6D defined pDC lineage commitment along the lymphoid branch. Transcriptional characterization of SiglecH+Ly6D+ precursors indicated that pDC development requires high expression of the transcription factor IRF8, whereas pDC identity relies on TCF4. RNA sequencing of IL-7R+ lymphoid and CDP-derived pDCs mirrored the heterogeneity of mature pDCs observed in single-cell analysis. Both mature pDC subsets are able to secrete type 1 interferons, but only myeloid-derived pDCs share with cDCs their ability to process and present antigen.

Where's Waldo: Identifying DCs within Mononuclear Phagocytes during Inflammation.

Dendritic cells (DCs) are antigen-presenting cells subdivided in specialized subsets. In this issue, Bosteels et al. challenge this concept, identifying a unique subset of inflammatory DCs characterized by hybrid myeloid features, capable of efficiently priming CD4+ as well as CD8+ T cells.

A distinct human cell type expressing MHCII and RORγt with dual characteristics of dendritic cells and type 3 innate lymphoid cells.

Recent studies have characterized various mouse antigen-presenting cells (APCs) expressing the lymphoid-lineage transcription factor RORγt (Retinoid-related orphan receptor gamma t), which exhibit distinct phenotypic features and are implicated in the induction of peripheral regulatory T cells (Tregs) and immune tolerance to microbiota and self-antigens. These APCs encompass Janus cells and Thetis cell subsets, some of which express the AutoImmune REgulator (AIRE). RORγt+ MHCII+ type 3 innate lymphoid cells (ILC3) have also been implicated in the instruction of microbiota-specific Tregs. While RORγt+ APCs have been actively investigated in mice, the identity and function of these cell subsets in humans remain elusive. Herein, we identify a rare subset of RORγt+ cells with dendritic cell (DC) features through integrated single-cell RNA sequencing and single-cell ATAC sequencing. These cells, which we term RORγt+ DC-like cells (R-DC-like), exhibit DC morphology, express the MHC class II machinery, and are distinct from all previously reported DC and ILC3 subsets, but share transcriptional and epigenetic similarities with DC2 and ILC3. We have developed procedures to isolate and expand them in vitro, enabling their functional characterization. R-DC-like cells proliferate in vitro, continue to express RORγt, and differentiate into CD1c+ DC2-like cells. They stimulate the proliferation of allogeneic T cells. The identification of human R-DC-like cells with proliferative potential and plasticity toward CD1c+ DC2-like cells will prompt further investigation into their impact on immune homeostasis, inflammation, and autoimmunity.

The carboxy terminus causes interfacial assembly of oleate hydratase on a membrane bilayer.

The soluble flavoprotein oleate hydratase (OhyA) hydrates the 9-cis double bond of unsaturated fatty acids. OhyA substrates are embedded in membrane bilayers; OhyA must remove the fatty acid from the bilayer and enclose it in the active site. Here, we show that the positively charged helix-turn-helix motif in the carboxy terminus (CTD) is responsible for interacting with the negatively charged phosphatidylglycerol (PG) bilayer. Super-resolution microscopy of Staphylococcus aureus cells expressing green fluorescent protein fused to OhyA or the CTD sequence shows subcellular localization along the cellular boundary, indicating OhyA is membrane-associated and the CTD sequence is sufficient for membrane recruitment. Using cryo-electron microscopy, we solved the OhyA dimer structure and conducted 3D variability analysis of the reconstructions to assess CTD flexibility. Our surface plasmon resonance experiments corroborated that OhyA binds the PG bilayer with nanomolar affinity and we found the CTD sequence has intrinsic PG binding properties. We determined that the nuclear magnetic resonance structure of a peptide containing the CTD sequence resembles the OhyA crystal structure. We observed intermolecular NOE from PG liposome protons next to the phosphate group to the CTD peptide. The addition of paramagnetic MnCl2 indicated the CTD peptide binds the PG surface but does not insert into the bilayer. Molecular dynamics simulations, supported by site-directed mutagenesis experiments, identify key residues in the helix-turn-helix that drive membrane association. The data show that the OhyA CTD binds the phosphate layer of the PG surface to obtain bilayer-embedded unsaturated fatty acids.

SLC7A8 is a key amino acids supplier for the metabolic programs that sustain homeostasis and activation of type 2 innate lymphoid cells.

Group 2 innate lymphoid cells (ILC2) are innate counterparts of T helper 2 (Th2) cells that maintain tissue homeostasis and respond to injuries through rapid interleukin (IL)-5 and IL-13 secretion. ILC2s depend on availability of arginine and branched-chain amino acids for sustaining cellular fitness, proliferation, and cytokine secretion in both steady state and upon activation. However, the contribution of amino acid transporters to ILC2 functions is not known. Here, we found that ILC2s selectively express Slc7a8, encoding a transporter for arginine and large amino acids. Slc7a8 was expressed in ILC2s in a tissue-specific manner in steady state and was further increased upon activation. Genetic ablation of Slc7a8 in lymphocytes reduced the frequency of ILC2s, suppressed IL-5 and IL-13 production upon stimulation, and impaired type 2 immune responses to helminth infection. Consistent with this, Slc7a8-deficient ILC2s also failed to induce cytokine production and recruit eosinophils in a model of allergic lung inflammation. Mechanistically, reduced amino acid availability due to Slc7a8 deficiency led to compromised mitochondrial oxidative phosphorylation, as well as impaired activation of mammalian target of rapamycin and c-Myc signaling pathways. These findings identify Slc7a8 as a key supplier of amino acids for the metabolic programs underpinning fitness and activation of ILC2s.

One size does not fit all: Methodological considerations and recommended solutions for intramuscular temperature assessment.

Intramuscular temperature kinetics can provide insightful information for exercise and environmental physiology research. However, currently, there are no consistent method descriptions or guidelines for muscle temperature assessment in the literature. Studies have reported a great variation in muscle temperature assessment, from 1.5 cm under the skin to 4 cm under the muscle fascia. Moreover, a large variation in body composition components among participants exacerbates this issue, changing the depth and the muscle to be tested. For instance, in young adults (25 ± 5 yrs), the thigh subcutaneous fat thickness can vary from 0.11 to 1.69 cm, and vastus lateralis thickness from 1.62 to 3.38 cm; in older adults (68.5 ± 3 yrs), subcutaneous fat thickness plus gastrocnemius medialis thickness can vary from 1.03 to 3.22 cm. This variation results in inconsistent resting muscle temperature profiles and muscle temperature kinetics during and after an exercise or environmental thermal stress interventions (hot or cold). Hence, one fixed size does not fit all. Standardization and consistency in muscle temperature assessment procedures across studies are required to allow a better understanding and translation of the influence of a given stressor (exercise or thermal) on muscle temperature kinetics. This methodological manuscript i) summarizes the differences in muscle temperature assessment procedures and techniques used across different studies, ii) discusses current concerns related to variations in intramuscular needle depth, and subcutaneous fat and muscle thickness when assessing muscle temperature, and iii) suggests a systematic and more robust approach, based on individual body composition characteristics, to be considered when assessing intramuscular temperature.

Intrinsic motor neuron excitability is increased after resistance training in older adults.

This study investigated the effects of high-intensity resistance training on estimates of the motor neuron persistent inward current (PIC) in older adults. Seventeen participants (68.5 ± 2.8 yr) completed a 2-wk nonexercise control period followed by 6 wk of resistance training. Surface electromyographic signals were collected with two 32-channel electrodes placed over soleus to investigate motor unit discharge rates. Paired motor unit analysis was used to calculate delta frequency (ΔF) as an estimate of PIC amplitudes during 1) triangular-shaped contractions to 20% of maximum torque capacity and 2) trapezoidal- and triangular-shaped contractions to 20% and 40% of maximum torque capacity, respectively, to understand their ability to modulate PICs as contraction intensity increases. Maximal strength and functional capacity tests were also assessed. For the 20% triangular-shaped contractions, ΔF [0.58-0.87 peaks per second (pps); P ≤ 0.015] and peak discharge rates (0.78-0.99 pps; P ≤ 0.005) increased after training, indicating increased PIC amplitude. PIC modulation also improved after training. During the control period, mean ΔF differences between 20% trapezoidal-shaped and 40% triangular-shaped contractions were 0.09-0.18 pps (P = 0.448 and 0.109, respectively), which increased to 0.44 pps (P < 0.001) after training. Also, changes in ΔF showed moderate to very large correlations (r = 0.39-0.82) with changes in peak discharge rates and broad measures of motor function. Our findings indicate that increased motor neuron excitability is a potential mechanism underpinning training-induced improvements in motor neuron discharge rate, strength, and motor function in older adults. This increased excitability is likely mediated by enhanced PIC amplitudes, which are larger at higher contraction intensities.NEW & NOTEWORTHY Resistance training elicited important alterations in soleus intrinsic motor neuronal excitability, likely mediated by enhanced persistent inward current (PIC) amplitude, in older adults. Estimates of PICs increased after the training period, accompanied by an enhanced ability to increase PIC amplitudes at higher contraction intensities. Our data also suggest that changes in PIC contribution to self-sustained discharging may contribute to increases in motor neuron discharge rates, maximal strength, and functional capacity in older adults after resistance training.

Increases in muscle temperature by hot water improve muscle contractile function and reduce motor unit discharge rates.

PURPOSE: Examine the effects of 42°C hot-water immersion on muscle contraction function and motor unit discharge rates. Voluntary and evoked contraction assessments were examined first with a concomitant increase in the core and muscle temperature, and thereafter with increased muscle temperature but cooled core temperature. METHODS: Fifteen participants (24.9 ± 5.6 years) performed neuromuscular assessments before, after, and ~15-min after either 90-min of 42°C (hot) or 36°C (control) water immersion. Maximal voluntary contraction (MVC) assessment of knee extension was performed along with surface electromyography (sEMG) (vastus lateralis and medialis [VL, VM]) and voluntary activation level (VAL). Resting evoked twitch was elicited for peak torque and time to peak torque analysis. In addition, the VL and VM motor unit discharge rates (MUDR) were measured. RESULTS: After hot-water immersion (core temperature ↑1°C; muscle temperature ↑2.4°C), MVC torque and VAL decreased (p < 0.05). The sEMG (VL and VM) and peak twitch torque did not change (p > 0.05), while time to peak torque decreased (p = 0.007). The VL and VM MUDR decreased, showing a time effect, after both water immersion conditions (36 and 42°C) (p > 0.001). Fifteen minutes after the hot-water immersion (core temperature at baseline; muscle temperature ↑1.4°C), MVC torque returned to baseline, but VAL remained lower. The sEMG (VL and VM) remained unchanged. Peak twitch torque increased (p < 0.002) and time to peak torque remained lower (p = 0.028). The MUDR remained lower after both water immersion conditions (p < 0.05). CONCLUSION: Increased core temperature evoked by 42°C hot-water immersion decreases MVC torque and VAL. However, a passive increase in muscle temperature improved evoked muscle contractile function (i.e., time to peak torque [after] and peak twitch torque [~15 min after]). Moreover, a passive increase in muscle temperature reduced the required MUDR to attain the same torque.

Potential role of passively increased muscle temperature on contractile function.

Declines in muscle force, power, and contractile function can be observed in older adults, clinical populations, inactive individuals, and injured athletes. Passive heating exposure (e.g., hot baths, sauna, or heated garments) has been used for health purposes, including skeletal muscle treatment. An acute increase in muscle temperature by passive heating can increase the voluntary rate of force development and electrically evoked contraction properties (i.e., time to peak twitch torque, half-relation time, and electromechanical delay). The improvements in the rate of force development and evoked contraction assessments with increased muscle temperature after passive heating reveal peripheral mechanisms' potential role in enhancing muscle contraction. This review aimed to summarise, discuss, and highlight the potential role of an acute passive heating stimulus on skeletal muscle cells to improve contractile function. These mechanisms include increased calcium kinetics (release/reuptake), calcium sensitivity, and increased intramuscular fluid.

Muscle temperature kinetics and thermoregulatory responses to 42 °C hot-water immersion in healthy males and females.

PURPOSE: To determine the vastus lateralis muscle temperature kinetics during and after passive heating, to exam the effect of sex on thermoregulatory responses, and the thermal safety and tolerance of the 42 °C hot-water immersion protocol. METHODS: Thirty participants (15 males, 15 females) underwent a 2 h 42 ºC hot-water immersion to the waist level. Vastus lateralis, rectal and skin temperature, thermal sensation, heart rate and blood pressure (BP) were measured during the passive heating and recovery period. Participant recovery was monitored until muscle temperature returned to baseline. RESULTS: Vastus lateralis temperature increased to a maximal value of 39.0 ± 0.11 °C (P < 0.001), reaching a plateau after ~ 83.5 min of hot-water immersion and returning to baseline after ~ 115.8 min of recovery. Despite the anthropometric differences between males and females (e.g., height, body mass, body fat %, and fat thickness; P < 0.05), thermoregulatory responses showed no differences between sexes (P > 0.05). No change was found in systolic BP (~ 117 mmHg; P = 0.061). Peak rectal temperature (38.8 ± 0.14 °C; P < 0.001), heart rate (~ 100 bpm; P < 0.001), and diastolic BP (↓ ~ 13 mmHg; P < 0.001) during the hot-water immersion indicated the safety of the protocol. While skin temperature (~ 35.4 °C; P < 0.001) and thermal sensation (~ 5.95 AU; P < 0.001) confirmed protocol tolerance. CONCLUSION: These data demonstrate lower-body 42 °C hot-water immersion to increase vastus lateralis temperature and plateau ~ 2.8 °C above baseline. This amplitude of muscle temperature change aligns with reported cellular adaptation and muscle growth. Thermal strain incurred from this protocol appears safe and tolerable, positioning it well for health-related prescription.

Physical exercise associated with vitamin D chronic supplementation reduces kidney injury induced by monosodium glutamate.

The aim was to evaluate the effects of chronic vitamin D (VD) supplementation associated with regular swimming over renal histomorphometric aspects in obese rats. Thirty Wistar male rats (5 days old) were used. Twenty four rats were given subcutaneous injections of monosodium glutamate (MSG; 4 g/kg), and six control rats were given an equimolar saline solution. At 21-days-old, the MSG-treated rats were randomly distributed among sedentary animals (S) and exercised (E, swimming; 3x/week). These groups were subdivided into groups orally supplemented with VD (12 µg/kg; 3x/week) or not supplemented (NS), totaling Five experimental groups (n = 6 rats/group): MSG, MSG-SVD, MSG-ENS, MSG-EVD and control groups. In MSG-obese rats, there was such as a decrease in the diameter of the, glomerular tuft, Bowman's capsule, Bowman's space areas, and renal cortical thickness, compared to the control group. In MSG-SVD, MSG-ENS, and MSG-EVD animals, there was an increase in the cortical thickness in relation to the MSG group. In MSG-ENS and MSG-EVD animals, there was a reduction of tubular degeneration in relation to the MSG group. We conclude that physical exercise associated with Vitamin D supplementation can prevent of renal injury, increasing the thickness of the renal cortex and decrease the tubular degeneration.

Effects of passive heating intervention on muscle hypertrophy and neuromuscular function: A preliminary systematic review with meta-analysis.

Passive heating has been therapeutically used to treat a range of health conditions. Further, this intervention presents as a potential exercise mimetic strategy showing acute and chronic effects on skeletal muscle adaptation and neuromuscular systems. This systematic review and meta-analysis aimed to synthesise the existing evidence on the effects of passive heating on muscle hypertrophy and neuromuscular function. Seven databases were searched (i.e., PubMed, Web of Science, Scopus, CINAHL, EMBASE, Cochrane, and SPORTDiscus) from 1937 to October 2019. Eligible studies included original papers using healthy animals or human samples (≥18 years; both sexes) that have used a control group or condition. Ten original articles were included in this review and four in the meta-analysis. The meta-analysis detected an increase in muscle mass in animal samples seven days after passive heating (I2 = 65%, P < 0.01). The systematic review showed preliminary evidence that repeated passive heating exposures may promote muscle hypertrophy in animals and humans. Moreover, augmented muscle strength (involuntary and voluntary) may be observed after long-term passive heating (animals and humans) and increases in corticospinal excitability in humans after a single passive heating session. Passive heating has shown some potential benefits for skeletal muscle mass gain and muscle force improvement. Therefore, it is plausible to suggest that passive heating might be a worthwhile alternative to be recommended as an exercise mimetic for those people who lack or are unable to complete sufficient exercise.

RING E3 mechanism for ubiquitin ligation to a disordered substrate visualized for human anaphase-promoting complex.

For many E3 ligases, a mobile RING (Really Interesting New Gene) domain stimulates ubiquitin (Ub) transfer from a thioester-linked E2∼Ub intermediate to a lysine on a remotely bound disordered substrate. One such E3 is the gigantic, multisubunit 1.2-MDa anaphase-promoting complex/cyclosome (APC), which controls cell division by ubiquitinating cell cycle regulators to drive their timely degradation. Intrinsically disordered substrates are typically recruited via their KEN-box, D-box, and/or other motifs binding to APC and a coactivator such as CDH1. On the opposite side of the APC, the dynamic catalytic core contains the cullin-like subunit APC2 and its RING partner APC11, which collaborates with the E2 UBCH10 (UBE2C) to ubiquitinate substrates. However, how dynamic RING-E2∼Ub catalytic modules such as APC11-UBCH10∼Ub collide with distally tethered disordered substrates remains poorly understood. We report structural mechanisms of UBCH10 recruitment to APC(CDH1) and substrate ubiquitination. Unexpectedly, in addition to binding APC11's RING, UBCH10 is corecruited via interactions with APC2, which we visualized in a trapped complex representing an APC(CDH1)-UBCH10∼Ub-substrate intermediate by cryo-electron microscopy, and in isolation by X-ray crystallography. To our knowledge, this is the first structural view of APC, or any cullin-RING E3, with E2 and substrate juxtaposed, and it reveals how tripartite cullin-RING-E2 interactions establish APC's specificity for UBCH10 and harness a flexible catalytic module to drive ubiquitination of lysines within an accessible zone. We propose that multisite interactions reduce the degrees of freedom available to dynamic RING E3-E2∼Ub catalytic modules, condense the search radius for target lysines, increase the chance of active-site collision with conformationally fluctuating substrates, and enable regulation.

Passive heat therapy: a promising preventive measure for people at risk of adverse health outcomes during heat extremes.

The world is experiencing increased frequency, duration, and severity of life-threatening heat extremes. Most hospitalizations and excess deaths during extreme heat events are associated with preexisting diseases in older adults. As climate change persists, the global population ages and the number of individuals with chronic diseases expands, more people are at risk of adverse health outcomes during extreme heat events. Therefore, proactive preventive measures are urgently needed to mitigate heat-related health risks within these populations. In this context, passive heat therapy (e.g., hot baths, saunas, and water-perfused suits) emerges as a promising countermeasure to improve physiological resilience to a warming planet. Passive heating improves cardiovascular function and overall health in older adults and individuals living with chronic diseases, offering the prospect of reducing cardiovascular strain during hotter days. Moreover, some studies suggest that passive heat therapy can be an effective strategy for heat acclimation (i.e., improved thermoregulation). This review describes the existing literature on the effects of passive heat therapy on cardiovascular and thermoregulatory responses in individuals with higher heat-related health risks and explores the use of passive heating as a strategy for heat acclimation to mitigate health risks during extreme heat events.NEW & NOTEWORTHY Passive heat therapy improves cardiovascular function and health in middle-aged and older adults living with or without chronic diseases. In addition, preliminary studies indicate that passive heat interventions can induce heat acclimation, improving thermoregulatory responses. Thus, passive heat therapy could serve as a preventive measure for people at risk of adverse health outcomes during extreme heat events, improving resilience to ongoing climate change.

Brain-Engrafted Monocyte-derived Macrophages from Blood and Skull-Bone Marrow Exhibit Distinct Identities from Microglia.

Microglia are thought to originate exclusively from primitive macrophage progenitors in the yolk sac (YS) and to persist throughout life without much contribution from definitive hematopoiesis. Here, using lineage tracing, pharmacological manipulation, and RNA-sequencing, we elucidated the presence and characteristics of monocyte-derived macrophages (MDMs) in the brain parenchyma at baseline and during microglia repopulation, and defined the core transcriptional signatures of brain-engrafted MDMs. Lineage tracing mouse models revealed that MDMs transiently express CD206 during brain engraftment as CD206 + microglia precursors in the YS. We found that brain-engrafted MDMs exhibit transcriptional and epigenetic characteristics akin to meningeal macrophages, likely due to environmental imprinting within the meningeal space. Utilizing parabiosis and skull transplantation, we demonstrated that monocytes from both peripheral blood and skull bone marrow can repopulate microglia-depleted brains. Our results reveal the heterogeneous origins and cellular dynamics of brain parenchymal macrophages at baseline and in models of microglia depletion.

The centrosomal protein FGFR1OP controls myosin function in murine intestinal epithelial cells.

Recent advances in human genetics have shed light on the genetic factors contributing to inflammatory diseases, particularly Crohn's disease (CD), a prominent form of inflammatory bowel disease. Certain risk genes associated with CD directly influence cytokine biology and cell-specific communication networks. Current CD therapies primarily rely on anti-inflammatory drugs, which are inconsistently effective and lack strategies for promoting epithelial restoration and mucosal balance. To understand CD's underlying mechanisms, we investigated the link between CD and the FGFR1OP gene, which encodes a centrosome protein. FGFR1OP deletion in mouse intestinal epithelial cells disrupted crypt architecture, resulting in crypt loss, inflammation, and fatality. FGFR1OP insufficiency hindered epithelial resilience during colitis. FGFR1OP was crucial for preserving non-muscle myosin II activity, ensuring the integrity of the actomyosin cytoskeleton and crypt cell adhesion. This role of FGFR1OP suggests that its deficiency in genetically predisposed individuals may reduce epithelial renewal capacity, heightening susceptibility to inflammation and disease.