F-53B-induced placental vascular endothelial dysfunction leads to intrauterine growth retardation of fetal mice

Preeclampsia is amultisystem disorder involving in inflammatory responses and metabolic dysfunction of maternal-fetal circulation. Recently, researchers found it threatens renal health of offspring in adulthood. Growing evidence indicated chronic kidney disease is associated with glomeruli deficiencies during intrauterine development. Our previous study showed placenta-derived exosomes from cord plasma with preeclampsia impede fetal glomerular vascularization, during which we postulate microRNAs may function as epigenetic switches for gene silencing of human glomerular endothelial cells. However, the specific miRNAs in placenta-derived exosomes engaged in glomerular vascularization remain unclear. Small RNA sequencing of placental-derived exosomes and bioinformatics analysis were applied to identify differentially expressed miRNAs, followed by real-time polymerase chain reaction for verification. Transient expression and inhibition of candidate miRNA were performed by transfection with chemically synthesized miRNA oligonucleotides. Functional assays of HGECs including cell proliferation assays, EDU assays, migration assays, tube formation assays and monolayer cell barrier permeability assays were performed after transfection. Further, dual luciferase assay was used to explore the target genes of candidate miRNA, followed by RT-qPCR, western blot and rescue assays. Antagomirs transfection of C57BL/6J fetal mice via Amniotic cavity injection in vivo and C57BL/6J fetal mice kidney explants culture in vitro were performed to evaluate number of glomeruli, renal development. Preeclampsia downregulates miR-199a-3p in placenta-derived exosomes from cord plasma. Suppression of endogenous miR-199a-3p in HGECs inhibits angiogenesis, proliferation, migration and permeability. A dual luciferase assay and rescue assays confirmed that miR-199a-3p targets PH domain leucine-rich repeat-containing protein phosphatase 2, regulating the phosphorylation of Akt serine/threonine kinase 1 (S473). C57BL/6J fetal mice with miR-199a-3p downregulation have low glomerulus counts and relative growth rate. miR-199a-3p in placenta-derived exosomes from cord plasma controls VEGF-induced glomerular angiogenesis. Moreover, it provides a distinct perspective for the mechanisms underlying the increased risk for renal disease in the offspring of preeclampsia patients via placenta-derived exosomal miRNAs.

Placental transfer of quercetin and its metabolites to fetuses in pregnant mice.

Multilineage potential induced by reconstruction of three-dimensional structures in mouse fetal brain cells.

Mouse model of prenatal valproic acid exposure: Effects on cortical morphogenesis and behavioral outcomes across environmental conditions.

Suppression of the long non-coding RNA, Malat1, enhances epithelial morphogenesis and differentiation in the fetal mouse submandibular gland

BackgroundMesenchymal stromal cells (MSCs) and their secretomes have been extensively studied for the treatment of various pulmonary diseases; however, consistent therapeutic benefits have yet to be demonstrated. The fundamental biology of lung-resident MSCs remains poorly understood.MethodsWe used a previously characterized fetal lung mesenchyme-specific reporter mouse model to initially identify lung MSC subpopulations based on their developmental origins. Bulk RNA-seq was then used to confirm mouse lung MSC differences at the transcriptomic level and to separate human lung MSC subsets. A variety of in vitro methods were used to measure cell proliferation, differentiation, adhesion, T cell modulation, alveolar epithelial cell growth.ResultsWe identified two distinct subsets of mouse lung resident MSCs: one originating from embryonic lung mesenchymal progenitors, and another with an as-yet undetermined tissue origin. While both subsets exhibit core MSC characteristics, transcriptomic analyses revealed distinct gene expression profiles, which were conserved in MSCs isolated from human lungs. These subsets also differed in key biological functions, including proliferation, adhesion, immunomodulation, and their capacity to support alveolar epithelial cell growth in both mouse and human systems.ConclusionsTwo different lung MSC subpopulations have been identified and characterized. These findings advance our understanding of the complexity of mesenchymal progenitors in lung injury repair and disease and may inform the development of effective stem cell-based therapies for pulmonary conditions.Supplementary InformationThe online version contains supplementary material available at 10.1186/s13287-025-04716-5.

Loss of fragile X messenger ribonucleoprotein (FMRP) causes fragile X syndrome (FXS), an inherited neurodevelopmental disorder resulting in intellectual disability and autism spectrum disorder; however, the molecular function of FMRP remains uncertain. Here, using cell lines and fibroblasts and induced pluripotent stem cell-derived neurons from healthy individuals and patients with FXS, we showed that FMRP regulates collided ribosomes by recruiting activating signal cointegrator 1 complex subunit 3 (ASCC3), an early-acting ribosome-associated quality control (RQC) factor to collided ribosomes, and either positively or negatively regulating translation, depending on transcript context. Disease-associated ASCC3 variants that perturbed ASCC3-FMRP interaction were also found to be defective in ribosome association and handling of collided ribosomes. In cells of a patient with FXS and the Fmr1 KO mouse model, ASCC3 abundance was reduced, and overexpression of ASCC3 in the brains of fetal Fmr1 KO mice promoted neuronal migration. In addition, CRISPR-mediated activation of ASCC3 by lateral ventricular injection of adeno-associated virus (AAV) ameliorated synaptic defects and improved locomotor activity, cognitive deficits, obsessive-compulsive-like behavior, and social interaction deficits after 1 month in 2-month-old Fmr1 KO mice compared with untreated Fmr1 KO controls. In conclusion, these data implicated FMRP in the handling of collided ribosomes to maintain protein homeostasis during neurodevelopment and synaptogenesis and demonstrated proof of concept that targeting RQC may offer alternative treatment strategies for FXS.

Although they exhibit limited regenerative ability of some tissues and organs shortly after birth or towards the end of fetal development, humans and laboratory mammals quickly transition to producing scar tissue for tissue repair. In contrast, spiny mice exhibit complex tissue regeneration as adults and provide a blueprint for how regeneration can occur throughout adulthood in mammals. Fibroblasts are key mediators of wound healing outcomes and prior work uncovered that cells from highly regenerative mammals (spiny mice and rabbits) exhibit extreme resistance to oxidative stress compared to those from non-regenerating laboratory mice and rats. Using a battery of cellular tests in primary ear pinna fibroblasts from highly regenerative and non-regenerative mammals, we find that cells from spiny mice and rabbits exhibit a baseline preference for glycolysis supporting a lower ROS-producing basal state. Uniquely, spiny mouse fibroblasts exhibit large, spherical, depolarized mitochondria similar to megamitochondria identified in pathological tissues. The megamitochondria phenotype was present across lifespan in ear pinna fibroblasts from fetal, young and old spiny mice. While rabbit, mouse and rat fibroblasts had polarized tubular mitochondrial networks typical of adult mammalian fibroblasts, isolated rabbit and spiny mice fibroblasts shared lower oxygen consumption efficiency even in the absence of a potential gradient. Taken together, our results support that a shared metabolic signature exists in stromal cells from highly regenerative mammals, although possibly driven by different mechanisms, to converge on a ROS-resistant phenotype which ultimately helps supports tissue regeneration.

Cartilage destruction is a characteristic of osteoarthritis, and intra-articular cartilage has poor spontaneous repair ability due to its non-nerve-free and non-vascular characteristics, so how to carry out cartilage repair and cartilage regeneration is a major problem in the current research of osteoarthritis. Mesenchymal stem cells (MSCs) are known to have multidirectional differentiation potential, while synovial mesenchymal stem cells (SMSCs) have been shown to have high proliferation rates, low immunogenicity, and strong chondrogenic differentiation potential in previous studies. There are stem cells in the articular cavity, and many methods have been applied to the study of intra-articular induction of stem cell cartilage differentiation, but there is still a lack of precise targets in this study. Nuclear factor of activated T cells 5 (NFAT5) plays a key role in a variety of cellular processes. Little is known about the role of NFAT5 in the differentiation and homeostasis of SMSCs. In this study, we found that the expression of NFAT5 in the cartilage proliferation region was significantly enhanced during the development of articular cartilage in fetal mice, and demonstrated that inhibition of NFAT5 expression by KRN2 had a significant effect on attenuating the chondrogenic differentiation of SMSCs. This finding suggests that NFAT5 may be a promising therapeutic target for cartilage regeneration.

Ovarian organogenesis requires the coordinated specification of supporting and steroidogenic cell lineages from multipotent coelomic epithelium (CE) progenitors. A longstanding question is whether the CE contains transcriptionally distinct, spatially organized progenitor subpopulations with predetermined lineage biases, or whether specification into supporting and steroidogenic lineages occurs only after delamination and integration into the bipotential gonad. The developmental origins of granulosa cells and the emergence of ovarian steroidogenic/stromal progenitors (SPs) also remain poorly defined. Here, we show that CE cells covering the fetal mouse ovary are transcriptionally heterogeneous and spatially organized into subdomains already primed toward supporting or steroidogenic fates. CE priming is dynamic, with transient coexistence of supporting- and steroidogenic-biased CE progenitors before resolving into a predominantly supporting-biased CE. Local delamination of these primed cells seeds intragonadal niches where pre-granulosa cells and SPs mirror the spatio-temporal arrangements of CE-primed progenitors. We further demonstrate a dual origin for the supporting lineage, with granulosa cells deriving from both the CE and supporting-like cells (SLCs). In parallel, we show that SPs arise from steroidogenic-primed CE cells, expand to represent 52% of ovarian somatic cells at birth, persist into adulthood and contribute to both theca and steroidogenic stromal cells. Together, these findings reveal transcriptionally and spatially distinct CE subpopulations that shape somatic lineage emergence with important implications for ovarian pathophysiology.

The mouse fetal intrauterine wound healing model is crucial and commonly used for investigating mechanisms and evaluating potential therapies for scarless skin regeneration compared to fibrotic healing. However, traditional intrauterine surgery remains technically challenging and understudied, which is associated with high maternal mortality and pregnancy loss, prompting us to refine the surgical protocol. Here, we report how the choice of surgical and mating procedure impact outcomes obtained. Pregnant mice underwent fetal surgery at embryonic days 15.5, 16.5 (E15.5, E16.5, scarless) and 18.5 (e18.5, fibrotic). Two surgical protocols were used: traditional method involved purse-string sutures, microsurgical scissors, amniotic fluid supplementation, and suture closure (Traditional); and our modified method omitting purse-string sutures, replacing scissors with needle puncture for uterine and fetal incisions, eliminating amniotic fluid supplementation, and employing skin staples for abdominal closure (Modified). The modified protocol significantly increased the likelihood of successful pregnancy, reduced operative time, decreased abortion rates, and enabled earlier modeling compared to the traditional method. At 48h, 7days, and 9days post-surgery, E15.5 wounds healed scarlessly, displaying regenerated hair follicles and organized collagen. Conversely, E18.5 wounds formed typical fibrotic scars, characterized by dense, disorganized collagen without hair follicles. The optimized surgical protocol presented here provides a simplified, reliable fetal mouse model with improved pregnancy success, reduced fetal loss, earlier implementation, and consistent phenotypic outcomes. This refined model enhances experimental efficiency, reproducibility, and animal welfare, having a major impact on mechanistic studies and therapeutic exploration for scarless skin regeneration.

Mass spectrometry imaging (MSI) is a label-free technique that enables the visualization of the spatial distribution of thousands of ions within biosamples. Data denoising is the computational strategy aimed at enhancing the MSI data quality, providing an effective alternative to experimental methods. However, due to the complex noise pattern inherent in MSI data and the difficulty in obtaining ground truth from noise-free data, achieving reliable denoised images remains challenging. In this study, we introduce De-MSI, a novel deep learning-based method specifically developed for denoising MSI data without ground truth. The core concept of De-MSI involves constructing the reliable training data set by leveraging prior knowledge of mass spectrometry from the noisy MSI data, followed by training a deep neural network to improve the data quality by removing the noise from the original images. De-MSI has demonstrated superior performance in improving data quality over the commonly used methods when applied to MALDI-acquired mouse fetus data sets on visual inspection. Quantitative evaluations further confirm its superiority, with De-MSI achieving a mean PSNR of 18.93 and a mean SSIM of 0.74 across all ion images. The ability of De-MSI to enhance data quality in high-resolution MSI data sets is confirmed using the mouse brain data set at a pixel size of 5 μm. Additionally, its application to denoise rat brain data sets using the DESI technique showcases its adaptability across different ionization methods. The proposed model holds significant promise as a vital tool for the efficient analysis and interpretation of MSI data.

In adult mammals and other highly developed animals, incomplete wound healing, scar formation, and fibrosis occur. No treatment for complete tissue regeneration is currently available. However, in mice, at up to 13 days of gestation, early embryonic wounds regenerate without visible scarring. In mouse fetuses, actin cable formation at the epidermal wound margin contributes to regeneration after wounding; however, the relationship between actin behavior and dermal regeneration or scar formation by myofibroblasts is unknown. In the present study, we observed actin dynamics in the wound dermis of mouse fetuses and investigated fibroblast and alpha-smooth muscle actin (α-SMA) properties involved in the switch between regeneration and scar formation in the dermis. In the wound healing process of mouse fetuses, actomyosin bundles develop and contract in a mesh-like pattern in different parts depending on the developmental stage, i.e., in the dermis of E13 (regeneration) and in the fascia of E15 and later (scar formation). Furthermore, in E13 dermal fibroblasts, α-SMA is present in the cytoplasm independently of actin, but in E15 and later myofibroblasts, TGFβ-1 stimulation causes the distribution of α-SMA and actin to coincide, and in E17, when dermal scarring occurs, α-SMA is expressed particularly in the nucleus. The results indicate that reticular contraction by actomyosin is involved in dermal regeneration, and that the discrepancy in the localization of actin and α-SMA in fibroblasts is necessary. The findings may contribute to effective wound regeneration therapy.

ObjectiveTo investigate the effects of zinc concentration on palatal development in fetal mice and its association with the aryl hydrocarbon receptor (AhR) signaling pathway.MethodsPregnant C57BL/6J mice were fed diets with varying zinc concentrations and randomly divided into a zinc-rich (ZR) group, a normal-zinc (NZ) group, and a zinc-deficient (ZD) group. Embryonic development was observed, and the expression levels of AhR signaling pathway-related factors were examined.ResultsNo cleft palate was observed in the ZR group or NZ group, whereas the zinc-deficient group exhibited a cleft palate incidence of 27.45%. Hematoxylin and Eosin (HE) staining results revealed failed palatal shelf contact and fusion in the ZD group, whereas complete fusion occurred in the ZR group and a normal medial edge epithelial formed in the NZ group. Results from Immunohistochemistry (IHC), qRT-PCR, and Western blot analyses collectively demonstrated that, compared to the NZ group, the ZR group exhibited significant upregulation (P < .05) in both mRNA and protein expression levels of AhR, HSP90, CYP1A1, and SP1, whereas the ZD group displayed significant downregulation (P < .05).ConclusionZinc concentration is intimately correlated with fetal mouse palatal development, where zinc deficiency may contribute to cleft palate formation through suppression of AhR signaling pathway, whereas zinc-rich conditions facilitate activation of this pathway. This study reveals the regulatory role of zinc-AhR signaling axis in palatal development through mouse models, offering novel theoretical insights into zinc deficiency-induced cleft palate pathogenesis and establishing a foundational framework for preventive interventions.

In Indonesia, several unregistered traditional Chinese medicines (UTCM) are still used to treat gastritis in pregnancy. On the other hand, the safety of medications during pregnancy remains unresolved. This study aims to investigate the teratogenic effect of UTCM containing Atractylodis lancea radix, Glycyrrhizae glabra radix, Rheum officinale rhizome, and Angelica dahurica radix on fetal morphology. A total of 80 pregnant mice were divided into four groups including the negative control (N), given vehicle, as well as treatment groups D1, D2, and D3 given a dose of 35.2 mg/kg BW, 70.5 mg/kg BW and 105.65 mg/kg BW daily respectively. The drugs were administered between the 6th and 15th days of organogenesis. On the 18th day of pregnancy, a laparotomy was conducted. The teratogenic effects were determined by measuring maternal and fetal body weight, the number of fetuses, and skeletal abnormalities in mouse fetuses, visualised with Alizarin solution. All data were analysed using ANOVA. The results showed that in all treatment groups, there was a substantial difference in maternal body weight, fetal number, and fetal body weight (p<0.05). There were open eyelids and clubfoot abnormalities observed in all groups, while 27 fetuses with haemorrhages were found in D3. All treatment groups had sternal, sacral, caudal, phalanges and metacarpal abnormalities. UTCM administration showed fetal development effects at the early stage of pregnancy in mice. The dose (D3) 105.65 mg/kg BW affected maternal weight gain and fetal skeletal ossification. UTCM administration D1 (35.2 mg/kg BW), D2 0.5 mg/kg BW), and D3 (105.65 mg/kg BW) had no maternal toxicity and fetal teratogenesis.

Pathological cardiac remodeling is associated with the reactivation of fetal genes, yet the extent of the heart's fetal gene program and its impact on proteome compositions remain incompletely understood. Here, using a proteome-wide protein ratio quantification strategy with mass spectrometry, we identified pervasive isoform usage shifts in fetal and postnatal mouse hearts, involving 145 pairs of highly homologous paralogs and alternative splicing-derived isoform proteins. Proteome-wide ratio comparisons readily rediscovered hallmark fetal gene signatures in muscle contraction and glucose metabolism pathways, while revealing what we believe to be previously undescribed isoform usage in mitochondrial and gene-expression-regulating proteins, including PPA1/PPA2, ANT1/ANT2, and PCBP1/PCBP2 switches. Paralogs with differential fetal usage tend to be evolutionarily recent, consistent with functional diversification. Alternative splicing adds another rich source of fetal isoform usage differences, involving PKM M1/M2, GLS1 KGA/GAC, PDLIM5 long/short, and other spliceoforms. When comparing absolute protein proportions, we observed a partial reversion toward fetal gene usage in pathological hearts. In summary, we present a ratiometric catalog of paralogs and spliceoform pairs in the cardiac fetal gene program. More generally, the results demonstrate the potential of applying the proteome-wide ratio test concept to discover new regulatory modalities beyond differential gene expression.

SPARC expression in the mouse decidua, placenta, and fetus: correlations with SPP1 expression.

Live autologous cultured fibroblasts improve sagging and wrinkles when injected into the dermis; however, their diversity and differences in efficacy depending on the skin layer to which they are transplanted remain unclear. This study investigated the behavior and effects of dermal- and fascial-derived fibroblasts from mouse fetuses on the dermis and fascia. The collagen-producing capacity of fibroblasts from the shallow (dermis) and deep (fascia) skin layers of fetal and adult C57BL/6-Tg(CAG-EGFP) mice was compared using western blotting and real-time polymerase chain reaction. Contractile capacity was investigated using a collagen contractility assay and an ex vivo mouse skin model. We transplanted these cells into the shallow and deep layers of the back skin of aged C57BL/6 mice and observed the engraftment rate and histological changes in the skin. The anti-inflammatory effects of these cells were also compared. Fascial fibroblasts from embryonic day 17 fetal mice exhibited higher collagen production and contractile capacity than papillary fibroblasts from adult mice. The transplanted fascial and dermal cells showed high engraftment rates in the deep and shallow (injected near the layer from which they were derived) layers, respectively. Collagen fiber density was highest when embryonic day 17 fascial fibroblasts were transplanted into the deep layer, and an anti-inflammatory effect was observed. Dermal fibroblast transplantation improves the phenotype of aging skin via collagen synthesis and contraction, with greater efficacy when transplanted into a layer consistent with the skin layer from which the primary cells are harvested.

Gene editing technologies have revolutionized therapies for numerous genetic diseases. However, in vivo gene editing hinges on identifying efficient delivery vehicles for editing in targeted cell types, a significant hurdle in fully realizing its therapeutic potential. A model system to rapidly evaluate systemic gene editing would advance the field. Here, we develop the GFP-on reporter mouse, which harbors a nonsense mutation in a genomic EGFP sequence correctable by adenine base editor (ABE) among other genome editors. The GFP-on system was validated using single and dual adeno-associated virus (AAV9) encoding ABE8e and sgRNA. Intravenous administration of AAV9-ABE8e-sgRNA into adult GFP-on mice results in EGFP expression consistent with the tropism of AAV9. Intrahepatic delivery of AAV9-ABE8e-sgRNA into GFP-on fetal mice restores EGFP expression in AAV9-targeted organs lasting at least six months post-treatment. The GFP-on model provides an ideal platform for high-throughput evaluation of emerging gene editing tools and delivery modalities.