title: Novel insights into capillary vessel basement membrane damage by snake venom hemorrhagic metalloproteinases: a biochemical and immunohistochemical study.
abstract: The hemorrhagic activity characteristic of viperid snake envenomations is due to the action of venom metalloproteinases (SVMPs) on the capillary vessel basement membrane (BM). This study compared the action of two SVMPs on BM in vitro (degradation of Matrigel) and in vivo (immunohistochemical assessment of BM markers in mouse gastrocnemius muscle). SVMPs BaP1 (belonging to the P-I class) and jararhagin (of the P-III class) had a similar proteolytic activity on azocasein and degraded Matrigel with a slightly different cleavage pattern, since BaP1 exerted a limited proteolysis of both laminin and nidogen, whereas jararhagin predominantly degraded nidogen. In contrast with this pattern of limited proteolysis of BM proteins observed in vitro, immunohistochemical analysis of laminin, nidogen and type IV collagen, as well as of the endothelial cell marker VEGFR-2, in the hemorrhagic areas in the muscle, revealed a pronounced reduction in the immunostaining of these three BM components, associated with a loss of the endothelial cell marker. BM of muscle fibers was affected to a lesser extent. In conclusion, in vitro results demonstrated that SVMPs induce a pattern of limited proteolysis on BM components. The drastic loss of these antigens in affected capillaries in vivo is likely to depend on the combination of limited proteolysis of BM and the action of hemodynamic biophysical forces, previously shown to play a role in SVMP-induced capillary damage, which may cause a mechanical disruption of BM structure.
authors.affiliation: Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica.
authors.forename: Teresa
authors.initials: T
authors.lastname: Escalante
authors.forename: John
authors.initials: J
authors.lastname: Shannon
authors.forename: Ana M
authors.initials: AM
authors.lastname: Moura-da-Silva
authors.forename: José María
authors.initials: JM
authors.lastname: Gutiérrez
authors.forename: Jay W
authors.initials: JW
authors.lastname: Fox
mesh_terms: D000818:Animals; D002196:Capillaries / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure*; D002478:Cells, Cultured; D003435:Crotalid Venoms / Q000008:administration & dosage*; D004305:Dose-Response Relationship, Drug; D016326:Extracellular Matrix Proteins / Q000378:metabolism*; D008666:Metalloendopeptidases / Q000008:administration & dosage*; D051379:Mice; D012910:Snake Venoms / Q000008:administration & dosage; D000097146:Bothrops jararaca Venom
chemical_list: D003435:Crotalid Venoms; D016326:Extracellular Matrix Proteins; D008666:Metalloendopeptidases; D012910:Snake Venoms; D000097146:Bothrops jararaca Venom; C104933:BaP1 metalloproteinase; C069317:hemorrhagic metalloproteinase