This protocol describes the preparation of pupae for laser ablation and live imaging of the notum (dorsal thorax). Because the pupa is stationary, it can be continuously live imaged for multiple days if desired, making it ideal for studying wound signaling and repair, from before laser ablation through wound closure. In this protocol, we demonstrate the processes of staging, partially dissecting, mounting, wounding, and live imaging the pupal notum, with the wounding occurring during the live imaging process. For complete details on the use and execution of this protocol, please refer to O'Connor et al. (2021b).
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http://dx.doi.org/10.1016/j.xpro.2022.101396 | DOI Listing |
Arterioscler Thromb Vasc Biol
January 2025
School of Life Science, Nantong Laboratory of Development and Diseases and Co-Innovation Center of Neuroregeneration, Nantong University, China.
Background: Sprouting blood vessels, reaching the aimed location, and establishing the proper connections are vital for building vascular networks. Such biological processes are subject to precise molecular regulation. So far, the mechanistic insights into understanding how blood vessels grow to the correct position are limited.
View Article and Find Full Text PDFMol Med
January 2025
Research Institute, National Cancer Center, Goyang-Si, Gyeonggi-Do, 10408, Republic of Korea.
Background: Double-strand breaks (DSBs) are primarily repaired through non-homologous end joining (NHEJ) and homologous recombination (HR). Given that DSBs are highly cytotoxic, PARP inhibitors (PARPi), a prominent class of anticancer drugs, are designed to target tumors with HR deficiency (HRD), such as those harboring BRCA mutations. However, many tumor cells acquire resistance to PARPi, often by restoring HR in HRD cells through the inactivation of NHEJ.
View Article and Find Full Text PDFACS Appl Bio Mater
January 2025
Department of Chemical Engineering, Indian Institute of Technology Bombay, Mumbai 400076, India.
Hemodialysis and bioartificial kidney (BAK), which mimic both physical and biological functions, can significantly impact chronic kidney disease (CKD) patients. Here we report on Hollow fiber membranes (HFMs) with enhanced separation of uremic toxins along with enhanced hemocompatibility and biocompatibility that also promote the growth of kidney cells. The improvement arises from the addition of titanium dioxide (0.
View Article and Find Full Text PDFNat Commun
January 2025
Department of Translational Neurobiology, National Institute of Neuroscience, National Center of Neurology and Psychiatry, Kodaira, Tokyo, 187-8502, Japan.
Caspases are known to mediate neuronal apoptosis during brain development. However, here we show that nonapoptotic activation of caspase-3 at presynapses drives microglial synaptic phagocytosis. Real-time observation and spatiotemporal manipulation of synaptic caspase-3 in the newly established, mouse-derived culture system demonstrate that increased neuronal activity triggers localized presynaptic caspase-3 activation, facilitating synaptic tagging by complements.
View Article and Find Full Text PDFCell Struct Funct
January 2025
Department of Pathology and Biology of Diseases, Graduate School of Medicine, Kyoto University.
Live imaging techniques have revolutionized our understanding of paracrine signaling, a crucial form of cell-to-cell communication in biological processes. This review examines recent advances in visualizing and tracking paracrine factors through four key stages: secretion from producing cells, diffusion through extracellular space, binding to target cells, and activation of intracellular signaling within target cells. Paracrine factor secretion can be directly visualized by fluorescent protein tagging to ligand, or indirectly by visualizing the cleavage of the transmembrane pro-ligands or plasma membrane fusion of endosomes comprising the paracrine factors.
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