title: Blood brain barrier precludes the cerebral arteries to intravenously-injected antisense oligonucleotide.
abstract: Alternative splicing of the ryanodine receptor subtype 3 (RyR3) produces a short isoform (RyR3S) able to negatively regulate the ryanodine receptor subtype 2 (RyR2), as shown in cultured smooth muscle cells from mice. The RyR2 subtype has a crucial role in the control of vascular reactivity via the fine tuning of Ca(2+) signaling to regulate cerebral vascular tone. In this study, we have shown that the inhibition of RyR3S expression by a specific antisense oligonucleotide (asRyR3S) was able to increase the Ca(2+) signals implicating RyR2 in cerebral arteries ex vivo. Moreover, we tried to inhibit the expression of RyR3S in vivo. The asRyR3S was complexed with JetPEI and injected intravenously coupled with several methods known to induce a blood brain barrier disruption. We tested solutions to induce osmotic choc (mannitol), inflammation (bacteria lipopolysaccharide and pertussis toxin), vasoconstriction or dilatation (sumatriptan, phenylephrine, histamine), CD73 activation (NECA) and lipid instability (Tween80). All tested technics failed to target asRyR3 in the cerebral arteries wall, whereas the molecule was included in hepatocytes or cardiomyocytes. Our results showed that the RyR3 alternative splicing could have a function in cerebral arteries ex vivo; however, the disruption of the blood brain barrier could not induce the internalization of antisense oligonucleotides in the cerebral arteries, in order to prove the function of RYR3 short isoform in vivo.
authors.affiliation: Univ. de Bordeaux, Institut des Maladies Neurodégénératives, UMR 5293, F-33000 Bordeaux, France; CNRS, Institut des Maladies Neurodégénératives, UMR 5293, F-33000 Bordeaux, France.
authors.forename: Raphael
authors.initials: R
authors.lastname: Boursereau
authors.affiliation: Univ. de Bordeaux, Institut des Maladies Neurodégénératives, UMR 5293, F-33000 Bordeaux, France; CNRS, Institut des Maladies Neurodégénératives, UMR 5293, F-33000 Bordeaux, France.
authors.forename: Arnaud
authors.initials: A
authors.lastname: Donadieu
authors.affiliation: University of Vermont, Department of Pharmacology, UVM College of Medicine, Burlington, VT, USA. Electronic address: Fabrice.Dabertrand@uvm.edu.
authors.forename: Fabrice
authors.initials: F
authors.lastname: Dabertrand
authors.affiliation: Centre de Neurophysique, Physiologie, Pathologie, CNRS UMR 8119, Faculté des Sciences fondamentales et Biomédicales, Université Paris Descartes, 45, rue des Saints-Pères, 75006 Paris, France. Electronic address: dubayle@biomedicale.univ-paris5.fr.
authors.forename: David
authors.initials: D
authors.lastname: Dubayle
authors.affiliation: Univ. de Bordeaux, Institut des Maladies Neurodégénératives, UMR 5293, F-33000 Bordeaux, France; CNRS, Institut des Maladies Neurodégénératives, UMR 5293, F-33000 Bordeaux, France. Electronic address: jean-luc.morel@u-bordeaux.fr.
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Morel
mesh_terms: D000818:Animals; D001692:Biological Transport; D001812:Blood-Brain Barrier / Q000378:metabolism*; D020013:Calcium Signaling / Q000235:genetics; D002536:Cerebral Arteries / Q000378:metabolism*; D007275:Injections, Intravenous; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009131:Muscle, Smooth, Vascular / Q000166:cytology; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage* / Q000235:genetics / Q000378:metabolism*; D020033:Protein Isoforms / Q000235:genetics; D019837:Ryanodine Receptor Calcium Release Channel / Q000235:genetics
chemical_list: D016376:Oligonucleotides, Antisense; D020033:Protein Isoforms; D019837:Ryanodine Receptor Calcium Release Channel
keywords: Blood brain barrier; Calcium signaling; Cerebral artery; Ryanodine receptor