MicroRNAs (miRNAs) are a class of small noncoding RNAs that modulate the expression of virtually all genes. miRNAs have been implicated in the regulation of a broad range of cellular and physiologic processes. Dysregulation of miRNAs has been found to be associated with a variety of diseases and disorders, including teratogenesis. One of the important strategies to reveal miRNA function in teratogenesis is the manipulation of their expression levels and patterns in specific regions in the developing embryos. The combination of microinjection technology with the whole embryo culture (WEC) system provides an excellent platform for the mechanistic study of the roles of miRNAs in teratogenesis. Here we describe an approach to investigating and manipulating miRNAs in cultured mouse embryos and present protocols for whole embryo culture, microinjection of miRNA mimics or inhibitors, validation of the efficacy of microRNA modulation in embryos, and assessment of the teratogen-induced apoptosis.
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http://dx.doi.org/10.1007/978-1-4939-7883-0_10 | DOI Listing |
STAR Protoc
January 2025
Department of Immunobiochemistry, Mannheim Institute for Innate Immunoscience (MI3), Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany. Electronic address:
Liver sinusoidal endothelial cells (LSECs) line the liver sinusoids and play a crucial role in liver function. Isolating LSECs is beneficial for their functional evaluation in vitro. Here, we provide a protocol for obtaining purified LSECs from mice via gradient centrifugation and magnetic cell sorting (MACS), yielding cells suitable for culture and downstream analyses.
View Article and Find Full Text PDFJ Cell Mol Med
January 2025
Department of Spine, Orthopaedic Center, Guangdong Second Provincial General Hospital, Jinan University, Guangzhou, China.
Osteogenic differentiation of bone marrow stem cells (BMSCs) is essential for bone tissue regeneration and repair. However, this process is often hindered by an unstable differentiation influenced by local microenvironmental factors. While small extracellular vesicles (sEVs) derived from osteogenically induced adipose mesenchymal stem cells (ADSCs) reportedly can promote osteogenic differentiation of BMSCs, the underlying molecular mechanisms remain incompletely understood.
View Article and Find Full Text PDFJ Pathol
January 2025
Department of Clinical Bio-resource Research and Development, Graduate School of Medicine, Kyoto University, Kyoto, Japan.
Spread through air spaces (STAS) is a histological finding of lung tumours where tumour cells exist within the air space of the lung parenchyma beyond the margin of the main tumour. Although STAS is an important prognostic factor, the pathobiology of STAS remains unclear. Here, we investigated the mechanism of STAS by analysing the relationship between STAS and polarity switching in vivo and in vitro.
View Article and Find Full Text PDFJ Vis Exp
December 2024
The David and Inez Myers Laboratory for Cancer Genetics, Department of Human Molecular Genetics and Biochemistry, Faculty of Health and Medical Sciences, School of Medicine, Tel Aviv University;
Cerebellar Purkinje cells (PCs) exhibit a unique interplay of high metabolic rates, specific chromatin architecture, and extensive transcriptional activity, making them particularly vulnerable to DNA damage. This necessitates an efficient DNA damage response (DDR) to prevent cerebellar degeneration, often initiated by PC dysfunction or loss. A notable example is the genome instability syndrome, ataxia-telangiectasia (A-T), marked by progressive PC depletion and cerebellar deterioration.
View Article and Find Full Text PDFDrug Des Devel Ther
January 2025
Department of Trauma Orthopedics, Affiliated Hospital of Jining Medical University, Jining, Shandong, 272007, People's Republic of China.
Purpose: Osteosarcoma (OS) is the most common malignant tumor associated with poor patient outcomes and a limited availability of therapeutic agents. Scutellarein (SCU) is a monomeric flavone bioactive compound with potent anti-cancer activity. However, the effects and mechanisms of SCU on the growth of OS remain unknown.
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