The reproductive and endocrine functions of the ovary involve spatially defined interactions among specialized cell populations. Despite the ovary’s importance in fertility and endocrine health, functional attributes of ovarian cells are largely uncharacterized. Here, we profiled >18,000 genes in 257 regions from the ovaries of two premenopausal donors to examine the functional units in the ovary. We also generated single-cell RNA sequencing data for 21,198 cells from three additional donors and identified four major cell types and four immune cell subtypes. Custom selection of sampling areas revealed distinct gene activities for oocytes, theca, and granulosa cells. These data contributed panels of oocyte-, theca-, and granulosa-specific genes, thus expanding the knowledge of molecular programs driving follicle development. Serial samples around oocytes and across the cortex and medulla uncovered previously unappreciated variation of hormone and extracellular matrix remodeling activities. This combined spatial and single-cell atlas serves as a resource for future studies of rare cells and pathological states in the ovary.

Objective: The transcription factor OTX2is implicated in ocular, craniofacial, and pituitary development. Design: We aimed to establish the contribution of OTX2 mutations in congenital hypopituitarism patients with/without eye abnormalities, study functional consequences, and establish OTX2 expression in the human brain, with a view to investigate the mechanism of action. Methods: We screened patients from the UK (n = 103), international centres (n = 24), and Brazil (n = 282); 145 were within the septo-optic dysplasia spectrum, and 264 had no eye phenotype. Transactivation ability of OTX2 variants was analysed in murine hypothalamic GT1-7 neurons. In situ hybridization was performed on human embryonic brain sections. Genetically engineered mice were generated with a series of C-terminal OTX2 variants. Results: Two chromosomal deletions and six haploinsufficient mutations were identified in individuals with eye abnormalities; an affected relative of one patient harboured the same mutation without an ocular phenotype. OTX2 truncations led to significant transactivation reduction. A missense variant was identified in another patient without eye abnormalities; however, studies revealed it was most likely not causative. In the mouse, truncations proximal to aa219 caused anophthalmia, while distal truncations and the missense variant were tolerated. During human embryogenesis, OTX2 was expressed in the posterior pituitary, retina, ear, thalamus, choroid plexus, and partially in the hypothalamus, but not in the anterior pituitary. Conclusions: OTX2 mutations are rarely associated with hypopituitarism in isolation without eye abnormalities, and may be variably penetrant, even within the same pedigree. Our data suggest that the endocrine phenotypes in patients with OTX2 mutations are of hypothalamic origin.

The human uterus is a complex and dynamic organ whose lining grows, remodels, and regenerates every menstrual cycle or upon tissue damage. Here, we applied single-cell RNA sequencing to profile more the 50,000 uterine cells from both the endometrium and myometrium of five healthy premenopausal individuals, and jointly analyzed the data with a previously published dataset from 15 subjects. The resulting normal uterus cell atlas contains more than 167K cells, representing the lymphatic endothelium, blood endothelium, stromal, ciliated epithelium, unciliated epithelium, and immune cell populations. Focused analyses within each major cell type and comparisons with subtype labels from prior studies allowed us to document supporting evidence, resolve naming conflicts, and propose a consensus annotation system of 39 subtypes. We release their gene expression centroids, differentially expressed genes, and messenger Ribonucleic Acid (mRNA) patterns of literature-based markers as a shared community resource. We identify multiple potential progenitor cells: compartment-wide progenitors for each major cell type and potential cross-lineage multipotent stromal progenitors that may replenish the epithelial, stromal, and endothelial compartments. Furthermore, many cell types and subtypes exhibit shifts in cell number and transcriptomes across different phases of the menstrual cycle. Finally, comparisons between premenopausal, postpartum, and postmenopausal samples revealed substantial alterations in tissue composition, particularly in the proportions of stromal, endothelial, and immune cells. The cell taxonomy and molecular markers we report here are expected to inform studies of both basic biology of uterine function and its disorders.

The quality of electron beams produced from plasma-based accelerators, i.e., normalized brightness and energy spread, has made transformative progress in the past several decades in both simulation and experiment. Recently, full-scale particle-in-cell (PIC) simulations have shown that electron beams with unprecedented brightness (${10}^{20}–{10}^{21}\mathrm{A}/{\mathrm{m}}^2/{\mathrm{rad}}^2$) and 0.1–1 MeV energy spread can be produced through controlled injection in a slowly expanding bubble that arises when a particle beam or laser pulse propagates in density gradient, or when a particle beam self-focuses in uniform plasma or has a superluminal flying focus. However, in previous simulations of work on self-injection triggered by an evolving laser driver in a uniform plasma, the resulting beams did not exhibit comparable brightnesses and energy spreads. Here, we demonstrate through the use of large-scale high-fidelity PIC simulations that a slowly expanding bubble driven by a laser pulse in a uniform plasma can indeed produce self-injected electron beams with similar brightness and energy spreads as for an evolving bubble driven by an electron beam driver. We consider laser spot sizes roughly equal to the matched spot sizes in a uniform plasma and find that the evolution of the bubble occurs naturally through the evolution of the laser. The effects of the electron beam quality on the choice of physical as well as numerical parameters, e.g., grid sizes and field solvers used in the PIC simulations are presented. It is found that this original and simplest injection scheme can produce electron beams with beam quality exceeding that of the more recent concepts.

Microplastics (MPs) are small plastic particles emerging as significant environmental pollutants and humans are ubiquitously exposed to MPs. MPs can be detected in human atherosclerotic plaques and are associated with a higher risk of cardiovascular disease (CVD) and stroke in humans. However, the impact of MP exposure on the cardiovascular system remains elusive. In the current study, we investigated the effects of exposure to MPs at an environmentally relevant dose on atherosclerosis development in male and female low-density lipoprotein receptor-deficient (LDLR^-/-) mice. LDLR^-/- mice were fed a semisynthetic low-fat (4.3 %), low-cholesterol (0.02 %) diet and exposed to 10 mg/kg body weight MPs via daily oral gavage for 9 weeks. Male and female LDLR^-/- mice fed the low-fat diet did not develop obesity phenotype and exposure to MPs did not affect adiposity and circulating lipid profiles in those lean mice. Intriguingly, MP exposure increased atherosclerotic lesion areas in the aortic root by 63 % (p = 0.0185) and brachiocephalic artery by 624 % (p = 0.0541) in male LDLR^-/- mice but did not significantly affect atherosclerosis in female mice. Single-cell RNA sequencing analysis of the whole aorta revealed that exposure to MPs affected the proportions and cellular processes of key atherogenesis-related cell types, especially endothelia cells. Consistently, MP exposure induced pro-atherogenic gene expression in murine primary endothelia cells and human endothelial cells in vitro. Our findings reveal the sex-specific atherogenic effects of MPs in vivo and provide mechanistic insights and new understanding of the impact of MPs on atherosclerosis development and CVD risk in humans.