Real-time four-dimensional (4D, xyzt) imaging of cultured avian embryos is an ideal method for investigating the complex movements of cells and tissues during early morphogenesis. While methods that transiently label cells, such as electroporation, are highly useful for dynamic imaging, they can also be limiting due to the number and type of cells that can be effectively targeted. In contrast, the heritable, stable, and long-term expression of a fluorescent protein driven by the exogenous promoter of a transgene overcomes these challenges. We have used lentiviral vectors to produce several novel transgenic quail lines that express fluorescent proteins either ubiquitously or in a cell-specific manner. These lines have proven to be useful models for dynamic imaging and analysis. Here, we provide detailed protocols for generating transgenic quail with the emphasis on producing high titer lentivirus , effectively introducing it into the early embryo and efficiently screening for G1 founder birds .
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http://dx.doi.org/10.1007/978-1-4939-7216-6_8 | DOI Listing |
J Urol
January 2025
Division of Urology, Department of Surgery, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA.
Purpose: Urinary incontinence (UI) is common in nulliparous female elite athletes, but underlying pathophysiology is inadequately understood. We examined urinary symptoms and associated pelvic floor anatomy and function in this population, hypothesizing that athletes with UI would exhibit pelvic floor findings seen in older incontinent women (e.g.
View Article and Find Full Text PDFActa Bioeng Biomech
September 2024
Xinjiang University, China.
: The purpose of this study was to investigate dynamic responses of Lenke1B+ spines of adolescent scoliosis patients to different frequencies. : Modal analysis, harmonic response analysis and transient dynamics of a full spine model inverted by the finite element method using Abaqus. : The first-order axial resonance frequency of 4.
View Article and Find Full Text PDFBiochemistry
January 2025
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, 9, Leontovycha 9, Kyiv 01054, Ukraine.
The self-assembly of fibrin is a vital process in blood clotting, primarily facilitated by the interactions between knobs "A" and "B" in the central E region of one molecule and the corresponding holes "a" and "b" in the peripheral D regions of two other fibrin molecules. However, the precise function of the interactions between knob "B" and hole "b" during fibrin polymerization remains a subject of ongoing debate. The present study focuses on investigating intermolecular interactions between knob "B" and hole "b".
View Article and Find Full Text PDFJ Physiol
January 2025
Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge, UK.
The mechanisms that drive placental dysfunction in pregnancies complicated by hypoxia and fetal growth restriction remain poorly understood. Changes to mitochondrial respiration contribute to cellular dysfunction in conditions of hypoxia and have been implicated in the pathoaetiology of pregnancy complications, such as pre-eclampsia. We used bespoke isobaric hypoxic chambers and a combination of functional, molecular and imaging techniques to study cellular metabolism and mitochondrial dynamics in sheep undergoing hypoxic pregnancy.
View Article and Find Full Text PDFJ Cell Biol
April 2025
Team R2D2: Retroviral RNA Dynamics and Delivery, IRIM, UMR9004, CNRS, University of Montpellier, Montpellier, France.
Retroviruses carry a genomic intron-containing RNA with a long structured 5'-untranslated region, which acts either as a genome encapsidated in the viral progeny or as an mRNA encoding the key structural protein, Gag. We developed a single-molecule microscopy approach to simultaneously visualize the viral mRNA and the nascent Gag protein during translation directly in the cell. We found that a minority of the RNA molecules serve as mRNA and that they are translated in a fast and efficient process.
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