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Vascular endothelial cell senescence is closely associated with cardiovascular disease. The deficiency of natriuretic peptide receptor A (NPRA), encoded by the natriuretic peptide receptor 1 (NPR1) gene, contributes to vascular endothelial aging, but the cause of its reduced expression is unclear. In this study, we performed reverse chromatin immunoprecipitation (R-ChIP) to analyze the binding proteins of the NPR1 promoter and found that nucleolar and coiled body phosphoprotein 1 (NOLC1) functions as a key regulator of NPRA transcription in endothelial cells. Similar to NPRA, NOLC1 is also expressed at reduced levels in senescent endothelial cells. Knockdown of NOLC1 decreased both NPRA mRNA and protein expression levels. Furthermore, inhibition of NOLC1 expression triggered cellular senescence hallmarks, including elevated p53/p21 levels, enhanced SA-β-gal activity, ROS accumulation, G0/G1 cell cycle arrest, and impaired migration. NPRA overexpression rescued senescence and dysfunction in NOLC1-deficient cells, restoring proliferative and migratory capacity. In conclusion, our findings demonstrate that nucleolar phosphoprotein NOLC1 is a key regulator of NPRA transcription in endothelial senescence.

Astrocytes are a major glial cell type, playing multiple roles in the development, function, and pathogenesis of the brain. Accordingly, neuronal–astrocyte communication is an important research area. However, because these cell types share the same developmental origin, selective manipulation of each cell type is needed for precise mechanistic understanding. Here, we generated two new Cre driver lines for selective gene manipulation in astrocytes: Slc7a10-IRES-Cre and Aldh1l1-IRES-Cre. An internal ribosome entry site (IRES)-Cre cassette was knocked-in to the 3′-untranslated region of the solute carrier family 7 member 10 (Slc7a10) or aldehyde dehydrogenase 1 family member L1 (Aldh1l1) locus without disrupting gene function. The Slc7a10-IRES-Cre line underwent highly selective recombination in astrocytes of the brain, apart from choroid plexus epithelial cells. The onset of recombination began after completion of differentiation in the astrocyte lineage. By contrast, the Aldh1l1-IRES-Cre line began recombination during astrocyte differentiation at early postnatal stages. Some leaky expression was observed in the oligodendrocyte lineage, probably due to early onset of Cre expression in an uncommitted glial progenitor state. Together, the combination of the two deleter lines with distinct temporal Cre expression patterns serves as valuable tools to understand the development and function of astrocytes.

The mammalian uterus contains glands in the endometrium that develop only or primarily after birth. In the mouse, endometrial glands govern post implantation pregnancy establishment via regulation of blastocyst implantation, stromal cell decidualization, and placental development. Here, we describe a new uterine glandular epithelium (GE) specific Cre recombinase mouse line that is useful to study endometrial gland development and function. Utilizing CRISPR-Cas9 genome editing, improved Cre recombinase (iCre) was inserted into the endogenous C-X-C motif chemokine ligand 15 (Cxcl15) gene. Cxcl15 mRNA, Cxcl15 protein, and Cxcl15-iCre recombinase activity were specific to the developing GE of the uterus. Cxcl15-iCre mice were crossed with floxed Foxa2 mice to conditionally delete Foxa2 specifically in the glands of the neonatal mouse uterus. This conditional deletion of Foxa2 in the developing neonatal uterus resulted in adult mice that lacked Foxa2 in the GE of the uterus, and the adult mice were infertile. The studies described here establish that Cxcl15-iCre mice are a valuable resource to elucidate and explore mechanisms regulating the development and function of glands in the uterus.

Secreted Frizzled-related proteins (SFRPs) are a small, ancient family of extracellular signaling pathway agonists and antagonists. In this study, we examined their expression patterns during regeneration in the holothurian Eupentacta fraudatrix using whole-mount in situ hybridization. Our results suggest that both genes are involved in proper water-vascular system formation and coelomic epithelium homeostasis in holothurians.

Tissue-specific gene manipulation using Cre/loxP or Flp/frt recombination systems is a cornerstone of genetically engineered mouse models. In this study, we aim to develop a novel hepatocyte-specific, tamoxifen-inducible Flp mouse line. BAC (bacterial artificial chromosome)-Alb(albumin)-FlpER2(estrogen receptor ligan binding domain) was developed by inserting IRES-FlpER2 cDNA between the translation stop codon and 3′-UTR of the mouse albumin gene in a bacterial artificial chromosome. Upon tamoxifen induction in mice crossed with reporter lines, western blotting, immunohistochemistry, immunofluorescence staining, and X-gal staining (5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside Staining) were used to verify the recombination efficiency and specificity of this mouse model. Recombination was highly efficient and specific in hepatocytes, with no recombination detected in intrahepatic cholangiocytes or other organs in this mouse model. We generated a new tamoxifen-induced hepatocyte-specific mouse model with highly efficient recombination specifically in hepatocytes, and this model can be used to generate tumor model lines.