title: Ligation of the jugular veins does not result in brain inflammation or demyelination in mice.
abstract: An alternative hypothesis has been proposed implicating chronic cerebrospinal venous insufficiency (CCSVI) as a potential cause of multiple sclerosis (MS). We aimed to evaluate the validity of this hypothesis in a controlled animal model. Animal experiments were approved by the institutional animal care committee. The jugular veins in SJL mice were ligated bilaterally (n = 20), and the mice were observed for up to six months after ligation. Sham-operated mice (n = 15) and mice induced with experimental autoimmune encephalomyelitis (n = 8) were used as negative and positive controls, respectively. The animals were evaluated using CT venography and (99m)Tc-exametazime to assess for structural and hemodynamic changes. Imaging was performed to evaluate for signs of blood-brain barrier (BBB) breakdown and neuroinflammation. Flow cytometry and histopathology were performed to assess inflammatory cell populations and demyelination. There were both structural changes (stenosis, collaterals) in the jugular venous drainage and hemodynamic disturbances in the brain on Tc99m-exametazime scintigraphy (p = 0.024). In the JVL mice, gadolinium MRI and immunofluorescence imaging for barrier molecules did not reveal evidence of BBB breakdown (p = 0.58). Myeloperoxidase, matrix metalloproteinase, and protease molecular imaging did not reveal signs of increased neuroinflammation (all p>0.05). Flow cytometry and histopathology also did not reveal increase in inflammatory cell infiltration or population shifts. No evidence of demyelination was found, and the mice remained without clinical signs. Despite the structural and hemodynamic changes, we did not identify changes in the BBB permeability, neuroinflammation, demyelination, or clinical signs in the JVL group compared to the sham group. Therefore, our murine model does not support CCSVI as a cause of demyelinating diseases such as multiple sclerosis.
authors.affiliation: Center for Systems Biology, Massachusetts General Hospital and Harvard Medical School, Richard B. Simches Research Center, Boston, Massachusetts, United States of America.
authors.forename: Wendy
authors.initials: W
authors.lastname: Atkinson
authors.forename: Reza
authors.initials: R
authors.lastname: Forghani
authors.forename: Gregory R
authors.initials: GR
authors.lastname: Wojtkiewicz
authors.forename: Benjamin
authors.initials: B
authors.lastname: Pulli
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Iwamoto
authors.forename: Takuya
authors.initials: T
authors.lastname: Ueno
authors.forename: Peter
authors.initials: P
authors.lastname: Waterman
authors.forename: Jessica
authors.initials: J
authors.lastname: Truelove
authors.forename: Rahmi
authors.initials: R
authors.lastname: Oklu
authors.forename: John W
authors.initials: JW
authors.lastname: Chen
mesh_terms: D000818:Animals; D001812:Blood-Brain Barrier; D020278:Demyelinating Autoimmune Diseases, CNS / Q000473:pathology*; D004195:Disease Models, Animal; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000473:pathology*; D005260:Female; D005434:Flow Cytometry; D007601:Jugular Veins / Q000473:pathology*; D008279:Magnetic Resonance Imaging; D051379:Mice; D008856:Microscopy, Fluorescence; D011877:Radionuclide Imaging