ABSTRACT
Developmental cell death plays an important role in the construction of functional neural circuits. In vertebrates, the canonical view proposes a selection of the surviving neurons through stochastic competition for target-derived neurotrophic signals, implying an equal potential for neurons to compete. Here we show an alternative cell fitness selection of neurons that is defined by a specific neuronal heterogeneity code. Proprioceptive sensory neurons that will undergo cell death and those that will survive exhibit different molecular signatures that are regulated by retinoic acid and transcription factors, and are independent of the target and neurotrophins. These molecular features are genetically encoded, representing two distinct subgroups of neurons with contrasted functional maturation states and survival outcome. Thus, in this model, a heterogeneous code of intrinsic cell fitness in neighboring neurons provides differential competitive advantage resulting in the selection of cells with higher capacity to survive and functionally integrate into neural networks.
Subject(s)
Models, Biological , Sensory Receptor Cells/cytology , Animals , Cell Death/drug effects , Cell Survival/drug effects , Chick Embryo , Core Binding Factor Alpha 3 Subunit/metabolism , Mice, Inbred C57BL , Proprioception/drug effects , Receptor, trkC/metabolism , Sensory Receptor Cells/drug effects , Signal Transduction/drug effects , Tretinoin/pharmacologyABSTRACT
A versatile method for decorating magnetic nanobeads (being composite materials from polymers and superparamagnetic nanoparticles) with silver nanoparticles of 3-6 nm size is presented. Control over the silver nanoparticle coverage at the nanobead surface is achieved by changing the reaction parameters. Moreover, the silver-decorated magnetic nanobeads (Ag-MNBs) are studied with respect to their in vitro cytotoxicity on two distinct tumour cell lineages under different parameters, i.e., dose, incubation time, magnetic field applied during the culturing, silver ion leakage, and colloidal stability. It is found that enhanced magnetically mediated cellular uptake and silver ion leakage from the Ag-MNBs surface are the main factors which affect the toxicity of the Ag-MNBs and allow the half-maximal inhibitory dose of silver to be reduced to only 32 µg mL(-1) . Furthermore, a synergic cytotoxicity induced by photo-activation of silver nanoparticles was also found.
Subject(s)
Antineoplastic Agents/chemistry , Magnetics , Metal Nanoparticles , Silver/chemistry , Antineoplastic Agents/metabolism , Cell Adhesion , Cell Line, Tumor , Flow Cytometry , Humans , Microscopy, Electron, Scanning , Microscopy, Electron, Transmission , Photochemical Processes , Silver/metabolismABSTRACT
We here exploit pH-responsive nanogels as carriers to deliver functional anti-GFP siRNA and superparamagnetic IONPs to HeLa-GFP cells. The siRNA release via pH-mediated endosomal escape is shown. The IONPs act first as magnetofection agents to boost cellular uptake and then as probes to track the release mechanism by electron microscopy.
Subject(s)
Drug Carriers/chemistry , Ferric Compounds/chemistry , Ferric Compounds/metabolism , Magnetic Phenomena , Nanoparticles/chemistry , RNA, Small Interfering/metabolism , Biological Transport , Gels , HeLa Cells , Humans , Hydrogen-Ion Concentration , Intracellular Space/metabolismABSTRACT
Superparamagnetic nanocarriers with tunable pH dependence of the surface charge are designed by a simple co-precipitation method. By exploiting electrostatic interactions, cationic or anionic payloads can be adsorbed and desorbed depending on the pH. On three different resulting nanocarrier systems, experiments of loading and release of gold nanoparticles as well as effective siRNA loading and in vitro delivery on human cells are performed.
Subject(s)
Drug Carriers/chemistry , Gold/chemistry , Magnetics , Metal Nanoparticles/chemistry , RNA, Small Interfering/administration & dosage , Anions/chemistry , Cations/chemistry , Drug Carriers/pharmacology , Humans , Hydrogen-Ion Concentration , Microscopy, Fluorescence , RNA, Small Interfering/pharmacology , Scattering, RadiationABSTRACT
In a previous article, we demonstrated the existence of fucosyl-containing O-glycans forms of nucleolin in bovine post-capillary venular endothelial cells (CVEC) and malignant cultured human A431 cells. The tool for this discovery was an antibody found to interact strongly and exclusively with nucleolin in total protein extracts. The antibody was originally raised against a mollusc glycoprotein and was demonstrated to be directed against its O-glycans, recently found to belong prevalently to the blood group H-antigen type with fucose linked in alpha1, 2 to galactose. Here, we show that si-RNA induced down-regulation of the expression of FUT1 and FUT2, the fucosyltransferases required for the biosynthesis of the terminal glycan motif Fucalpha-2-Galbeta-R, reduced expression of the fucosylated nucleolin glycoforms and their exposure at the cell surface in CVEC. Treatment of the cells with FUT1/2 siRNA also reduced their ability to bind and internalize endostatin and their adhesion efficiency and inhibited cell growth. Expression of FUT1, FUT2, and FUT6 was also analyzed in serum-stimulated versus serum-starved cells and in cells treated with FUT1 and FUT2 siRNA. A reduced expression of fucosylated nucleolin and inhibition of cell growth by suppressing FUT1/2 expression was also tested and shown to be exhibited in human A431 cells.