Nogo-A is secreted in extracellular vesicles, occurs in blood and can influence vascular permeability

Nogo-A is a transmembrane protein with multiple functions in the central nervous system (CNS), including restriction of neurite growth and synaptic plasticity. Thus far, Nogo-A has been predominantly considered a cell contact-dependent ligand signaling via cell surface receptors. Here, we show that Nogo-A can be secreted by cultured cells of neuronal and glial origin in association with extracellular vesicles (EVs). Neuron- and oligodendrocyte-derived Nogo-A containing EVs inhibited fibroblast spreading, and this effect was partially reversed by Nogo-A receptor S1PR2 blockage. EVs purified from HEK cells only inhibited fibroblast spreading upon Nogo-A over-expression. Nogo-A-containing EVs were found in vivo in the blood of healthy mice and rats, as well as in human plasma. Blood Nogo-A concentrations were elevated after acute stroke lesions in mice and rats. Nogo-A active peptides decreased barrier integrity in an in vitro blood-brain barrier model. Stroked mice showed increased dye permeability in peripheral organs when tested 2 weeks after injury. In the Miles assay, an in vivo test to assess leakage of the skin vasculature, a Nogo-A active peptide increased dye permeability. These findings suggest that blood borne, possibly EV-associated Nogo-A could exert long-range regulatory actions on vascular permeability

A novel hybrid promoter responsive to pathophysiological and pharmacological regulation

The aim of this study was to construct a promoter containing DNA motifs for an endogenous transcription factor associated with inflammation along with motifs for pharmacological regulation factors. We demonstrate in transfected cells that expression of a gene of interest is induced by hypoxic conditions or through pharmacological induction, and also show pharmacological repression. In vivo studies utilised electroporation of plasmid to mouse paws, a delivery method shown to be effective by bioluminescence imaging. For gene therapy, the promoter was used to drive expression of IL-1Ra in a paw inflammation model with therapeutic effect observed which was further enhanced when the promoter was additionally induced with a pharmacological activator. One of the most important observations from this study was that promoter induction by hypoxia or inflammation could be prevented by the pharmacological repressor in the absence of doxycycline. These studies demonstrate that hybrid promoters enable pharmacological adjustment to the pathophysiological level of gene expression and, importantly, that they allow termination of gene expression even in the presence of pathophysiological stimuli

Clinicopathologic effects of mutant GUCY2D in Leber congenital amaurosis

To study the retinal degeneration in an 11 1 2 -year-old patient with Leber congenital amaurosis (LCA) caused by mutation in GUCY2D. Comparative human tissue study. Two subjects with LCA; postmortem eye from one LCA patient and three normal donors. Clinical and visual function studies were performed between the ages of 6 and 10 years in the LCA eye donor and at age 6 in an affected sibling. Genomic DNA was screened for mutations in known LCA genes. The retina of the 11 1 2 -year-old subject with LCA was compared with normal retinas from donors age 3 days, 18 years, and 53 years. The tissues were processed for histopathologic studies and immunofluorescence with retinal cell-specific antibodies. Vision in both siblings at the ages examined was limited to severely impaired cone function. Mutation in the GUCY2D gene was identified in both siblings. Histopathologic study revealed rods and cones without outer segments in the macula and far periphery. The cones formed a monolayer of cell bodies, but the rods were clustered and had sprouted neurites in the periphery. Rods and cones were not identified in the midperipheral retina. The inner nuclear layer appeared normal in thickness throughout the retina, but ganglion cells were reduced in number. An 11 1 2 -year-old subject with LCA caused by mutant GUCY2D had only light perception but retained substantial numbers of cones and rods in the macula and far periphery. The finding of numerous photoreceptors at this age may portend well for therapies designed to restore vision at the photoreceptor level

Nogo-A is secreted in extracellular vesicles, occurs in blood and can influence vascular permeability

Nogo-A is a transmembrane protein with multiple functions in the central nervous system (CNS), including restriction of neurite growth and synaptic plasticity. Thus far, Nogo-A has been predominantly considered a cell contact-dependent ligand signaling via cell surface receptors. Here, we show that Nogo-A can be secreted by cultured cells of neuronal and glial origin in association with extracellular vesicles (EVs). Neuron- and oligodendrocyte-derived Nogo-A containing EVs inhibited fibroblast spreading, and this effect was partially reversed by Nogo-A receptor S1PR2 blockage. EVs purified from HEK cells only inhibited fibroblast spreading upon Nogo-A over-expression. Nogo-A-containing EVs were found in vivo in the blood of healthy mice and rats, as well as in human plasma. Blood Nogo-A concentrations were elevated after acute stroke lesions in mice and rats. Nogo-A active peptides decreased barrier integrity in an in vitro blood-brain barrier model. Stroked mice showed increased dye permeability in peripheral organs when tested 2 weeks after injury. In the Miles assay, an in vivo test to assess leakage of the skin vasculature, a Nogo-A active peptide increased dye permeability. These findings suggest that blood borne, possibly EV-associated Nogo-A could exert long-range regulatory actions on vascular permeability.ISSN:0271-678XISSN:1559-701

sj-pdf-1-jcb-10.1177_0271678X231216270 - Supplemental material for Nogo-A is secreted in extracellular vesicles, occurs in blood and can influence vascular permeability

Supplemental material, sj-pdf-1-jcb-10.1177_0271678X231216270 for Nogo-A is secreted in extracellular vesicles, occurs in blood and can influence vascular permeability by Ruslan Rust, Mea M Holm, Matteo Egger, Oliver Weinmann, Daniёlle van Rossum, Fruzsina R Walter, Ana Raquel Santa-Maria, Lisa Grönnert, Michael A Maurer, Simon Kraler, Alexander Akhmedov, Rose Cideciyan, Thomas F Lüscher, Maria A Deli, Inge K Herrmann and Martin E Schwab in Journal of Cerebral Blood Flow & Metabolism</p

sj-pdf-2-jcb-10.1177_0271678X231216270 - Supplemental material for Nogo-A is secreted in extracellular vesicles, occurs in blood and can influence vascular permeability

Supplemental material, sj-pdf-2-jcb-10.1177_0271678X231216270 for Nogo-A is secreted in extracellular vesicles, occurs in blood and can influence vascular permeability by Ruslan Rust, Mea M Holm, Matteo Egger, Oliver Weinmann, Daniёlle van Rossum, Fruzsina R Walter, Ana Raquel Santa-Maria, Lisa Grönnert, Michael A Maurer, Simon Kraler, Alexander Akhmedov, Rose Cideciyan, Thomas F Lüscher, Maria A Deli, Inge K Herrmann and Martin E Schwab in Journal of Cerebral Blood Flow & Metabolism</p