ABSTRACT
Condensing peptide-DNA complexes have great potential as nonviral agents for gene delivery. To date, however, such complexes have given transfection activities greatly inferior to adenovirus and somewhat inferior to cationic lipid-DNA complexes, even for cell lines and primary cells in vitro. We report here the identification of a novel condensing peptide, CL22, which forms DNA complexes that efficiently transfect many cell lines, as well as primary dendritic and endothelial cells. We report studies with sequence and structure variants that define some properties of the peptide that contribute to efficient transfection. We demonstrate that the superior transfection activity of CL22 compared with other DNA condensing peptides is conferred at a step after uptake of the complexes into cells. We show that CL22-DNA complexes have transfection activity that is at least equivalent to the best available nonviral agents.
Subject(s)
Peptide Fragments/genetics , Peptides/genetics , Transfection/methods , Amino Acid Sequence , Animals , Cell Culture Techniques , DNA/genetics , Dendritic Cells/metabolism , Endothelium, Vascular/cytology , Genetic Vectors , Humans , Mice , Molecular Sequence Data , Peptide Fragments/chemistry , Peptides/chemistry , Tumor Cells, CulturedABSTRACT
Subcellular localisation of xanthine oxidoreductase (XOR) was determined by indirect immunofluorescence using confocal microscopy in human endothelial and epithelial cell lines and in primary cultures of human umbilical vein endothelial cells. XOR was diffusely distributed throughout the cytoplasm but with higher intensity in the perinuclear region. In non-permeabilised cells, XOR was clearly seen to be asymmetrically located on the outer surfaces, showing, in many cases, a higher intensity on those faces apposed by closely neighbouring cells. Such specific distribution suggests a functional role for the enzyme in cell-cell interactions, possibly involving signalling via reactive oxygen species.
Subject(s)
Endothelium/enzymology , Epithelial Cells/enzymology , Xanthine Oxidase/metabolism , Cell Line , Cell Membrane/enzymology , Cytoplasm/enzymology , Endothelium/cytology , Endothelium, Vascular/cytology , Endothelium, Vascular/enzymology , Extracellular Space/enzymology , Humans , Umbilical VeinsABSTRACT
To enter vitellogenesis normally the females must pass through a period marked by a weak daily lighting which, it appears, triggers a determined physiological condition. If this state is not reached, vitellogenesis is impossible; on the contrary, if this state is reached, vitellogenesis is then accelerated by a long photophase. The results are discussed.
Subject(s)
Brachyura/physiology , Light , Reproduction , Animals , Female , Oogenesis , Ovary/physiology , Seasons , Sex CharacteristicsABSTRACT
The ovaries of females kept at 15 degrees C in the beginning of autumn, develop the next spring but in most of the crabs maintained at 25 degrees C these glands are still in previtellogenesis at that time. On the contrary, the ovarian development of eyestalkless animals is favoured by a high temperature. The inhibiting pedoncular hormone would then be ordered by the temperature. From the results obtained, it seems that the ovary does not order the appearance of sexual periodic characters.
