title: "In vivo cryotechnique" in combination with replica immunoelectron microscopy for caveolin in smooth muscle cells.
abstract: A novel "in vivo cryotechnique" with replica immunoelectron microscopy was developed for detecting caveolin localization on replica membranes prepared directly from living smooth muscle cells. After quick-freezing mouse duodenal walls by our "in vivo cryotechnique", the specimens were prepared for freeze-fracture and deep-etch replica membranes. Then they were treated with 5% SDS and 0.5% collagenase to keep some antigens on the replica membranes. The immunogold method could be used to clarify the localization of the caveolin antigen in relation to three-dimensional ultrastructures of living smooth muscle cells. Our new cryotechnique can provide native organization of functional molecules in living cells.
authors.affiliation: Department of Anatomy, Yamanashi Medical University, Tamaho, Japan.
authors.forename: I
authors.initials: I
authors.lastname: Takayama
authors.forename: N
authors.initials: N
authors.lastname: Terada
authors.forename: T
authors.initials: T
authors.lastname: Baba
authors.forename: H
authors.initials: H
authors.lastname: Ueda
authors.forename: Y
authors.initials: Y
authors.lastname: Kato
authors.forename: Y
authors.initials: Y
authors.lastname: Fujii
authors.forename: S
authors.initials: S
authors.lastname: Ohno
mesh_terms: D000818:Animals; D051242:Caveolin 1; D022461:Caveolins; D004386:Duodenum / Q000378:metabolism / Q000648:ultrastructure; D005613:Freeze Etching; D005614:Freeze Fracturing; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D016253:Microscopy, Immunoelectron; D009130:Muscle, Smooth / Q000378:metabolism* / Q000648:ultrastructure; D016628:Replica Techniques
chemical_list: C495541:Cav1 protein, mouse; D051242:Caveolin 1; D022461:Caveolins; D008565:Membrane Proteins