In-vivo microendoscopy in animal models became a groundbreaking technique in neuroscience that rapidly expands our understanding of the brain.Emerging hair-thin endoscopes based on multimode fibres are now opening up ...In-vivo microendoscopy in animal models became a groundbreaking technique in neuroscience that rapidly expands our understanding of the brain.Emerging hair-thin endoscopes based on multimode fibres are now opening up the prospect of ultra-minimally invasive neuroimaging of deeply located brain structures.Complementing these advancements with methods of functional imaging and optogenetics,as well as extending its applicability to awake and motile animals constitute the most pressing challenges for this technology.Here we demonstrate a novel fibre design capable of both,high-resolution imaging in immobilised animals and bending-resilient optical addressing of neurons in motile animals.The optimised refractive index profile and the probe structure allowed reaching a spatial resolution of 2μm across a 230μm field of view for the initial layout of the fibre.Simultaneously,the fibre exhibits negligible cross-talk between individual inner-cores during fibre deformation.This work provides a technological solution for imaging-assisted spatially selective photo-activation and activity monitoring in awake and freely moving animal models.展开更多
To screen for additional treatment targets against tongue cancer, we evaluated the contributions of extracellular signal-related kinase(ERK), AKT and ezrin in cancer development. Immunohistochemical staining showed th...To screen for additional treatment targets against tongue cancer, we evaluated the contributions of extracellular signal-related kinase(ERK), AKT and ezrin in cancer development. Immunohistochemical staining showed that ERK and ezrin expressions were significantly higher in invasive squamous cell carcinoma than in carcinoma in situ. To investigate the roles of ERK and ezrin in cancer development, we used the non-woven silica fibre sheet Cellbedwith a structure resembling the loose connective tissue morphology in a novel 3 D culture system. We confirmed that the 3 D system using CellbedTMaccurately mimicked cancer cell morphology in vivo. Furthermore, cell projections were much more apparent in 3 D-cultured tongue cancer cell lines than in 2 D cultures. Typically, under conventional 2 D culture conditions, F-actin and cortactin are colocalized in the form of puncta within cells.However, in the 3 D-cultured cells, colocalization was mainly observed at the cell margins, including the projections. Projections containing F-actin and cortactin colocalization were predicted to be invadopodia. Although suppressing ezrin expression with small interfering RNA transfection caused no marked changes in morphology, cell projection formation was decreased, and the tumour thickness in vertical sections after 3 D culture was markedly decreased after suppressing ERK activity because both the invasion ability and proliferation were inhibited. An association between cortactin activation as well as ERK activity and invadopodia formation was detected. Our novel 3 D culture systems using Cellbed? are simple and useful for in vitro studies before conducting animal experiments. ERK contributes to tongue cancer development by increasing both cancer cell proliferation and migration via cortactin activation.展开更多
基金The authors acknowledge the support from the European Research Council(724530),Ministry of Education,Youth and Sports(CZ.02.1.01/0.0/15_003/0000476)Thüringer Ministerium für Wirtschaft,the European Regional Development Fund(CZ.02.1.01/0.0/15_003/0000476)+1 种基金Wissenschaft und Digitale Gesellschaft,the Federal Ministry of Education and Research,Germany(BMBF)the Thüringer Aufbaubank.
文摘In-vivo microendoscopy in animal models became a groundbreaking technique in neuroscience that rapidly expands our understanding of the brain.Emerging hair-thin endoscopes based on multimode fibres are now opening up the prospect of ultra-minimally invasive neuroimaging of deeply located brain structures.Complementing these advancements with methods of functional imaging and optogenetics,as well as extending its applicability to awake and motile animals constitute the most pressing challenges for this technology.Here we demonstrate a novel fibre design capable of both,high-resolution imaging in immobilised animals and bending-resilient optical addressing of neurons in motile animals.The optimised refractive index profile and the probe structure allowed reaching a spatial resolution of 2μm across a 230μm field of view for the initial layout of the fibre.Simultaneously,the fibre exhibits negligible cross-talk between individual inner-cores during fibre deformation.This work provides a technological solution for imaging-assisted spatially selective photo-activation and activity monitoring in awake and freely moving animal models.
文摘To screen for additional treatment targets against tongue cancer, we evaluated the contributions of extracellular signal-related kinase(ERK), AKT and ezrin in cancer development. Immunohistochemical staining showed that ERK and ezrin expressions were significantly higher in invasive squamous cell carcinoma than in carcinoma in situ. To investigate the roles of ERK and ezrin in cancer development, we used the non-woven silica fibre sheet Cellbedwith a structure resembling the loose connective tissue morphology in a novel 3 D culture system. We confirmed that the 3 D system using CellbedTMaccurately mimicked cancer cell morphology in vivo. Furthermore, cell projections were much more apparent in 3 D-cultured tongue cancer cell lines than in 2 D cultures. Typically, under conventional 2 D culture conditions, F-actin and cortactin are colocalized in the form of puncta within cells.However, in the 3 D-cultured cells, colocalization was mainly observed at the cell margins, including the projections. Projections containing F-actin and cortactin colocalization were predicted to be invadopodia. Although suppressing ezrin expression with small interfering RNA transfection caused no marked changes in morphology, cell projection formation was decreased, and the tumour thickness in vertical sections after 3 D culture was markedly decreased after suppressing ERK activity because both the invasion ability and proliferation were inhibited. An association between cortactin activation as well as ERK activity and invadopodia formation was detected. Our novel 3 D culture systems using Cellbed? are simple and useful for in vitro studies before conducting animal experiments. ERK contributes to tongue cancer development by increasing both cancer cell proliferation and migration via cortactin activation.
