title: A systematic approach to assess locoregional differences in angiogenesis.
abstract: Skeletal muscle tissue differs with regard to the abundance of glycolytic and oxidative fiber types. In this context, capillary density has been described to be higher in muscle tissue with more oxidative metabolism as compared to that one with more glycolytic metabolism, and the highest abundance of capillaries has been found in boneward-oriented moieties of skeletal muscle tissue. Importantly, capillary formation is often analyzed as a measure for angiogenesis, a process that describes neo-vessel formation emanating from preexisting vessels, occurring, i.e., after arterial occlusion. However, a standardized way for investigation of calf muscle capillarization after surgically induced unilateral hind limb ischemia in mice, especially considering these locoregional differences, has not been provided so far. In this manuscript, a novel, methodical approach for reliable analysis of capillary density was established using anatomic-morphological reference points, and a software-assisted way of capillary density analysis is described. Thus, the systematic approach provided conscientiously considers intra-layer differences in capillary formation and therefore guarantees for a robust, standardized analysis of capillary density as a measure for angiogenesis. The significance of the methodology is further supported by the observation that capillary density in the calf muscle layers analyzed negatively correlates with distal lower limb perfusion measured in vivo.
authors.affiliation: Institut für Pharmakologie und Klinische Pharmakologie, Universitätsklinikum Düsseldorf, Heinrich-Heine-Universität Düsseldorf, Universitätsstraße 1, 40225, Düsseldorf, Germany.
authors.forename: T
authors.initials: T
authors.lastname: Driesen
authors.affiliation: Klinik für Kardiologie, Pneumologie und Angiologie, Universitätsklinikum Düsseldorf, Moorenstraße 5, 40225 Düsseldorf, Germany.
authors.forename: D
authors.initials: D
authors.lastname: Schuler
authors.affiliation: Institut für Pharmakologie und Klinische Pharmakologie, Universitätsklinikum Düsseldorf, Heinrich-Heine-Universität Düsseldorf, Universitätsstraße 1, 40225, Düsseldorf, Germany.
authors.forename: R
authors.initials: R
authors.lastname: Schmetter
authors.affiliation: Klinik für Kardiologie, Pneumologie und Angiologie, Universitätsklinikum Düsseldorf, Moorenstraße 5, 40225 Düsseldorf, Germany.
authors.forename: C
authors.initials: C
authors.lastname: Heiss
authors.affiliation: Klinik für Kardiologie, Pneumologie und Angiologie, Universitätsklinikum Düsseldorf, Moorenstraße 5, 40225 Düsseldorf, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Kelm
authors.affiliation: Institut für Pharmakologie und Klinische Pharmakologie, Universitätsklinikum Düsseldorf, Heinrich-Heine-Universität Düsseldorf, Universitätsstraße 1, 40225, Düsseldorf, Germany.
authors.forename: J W
authors.initials: JW
authors.lastname: Fischer
authors.affiliation: Institut für Pharmakologie und Klinische Pharmakologie, Universitätsklinikum Düsseldorf, Heinrich-Heine-Universität Düsseldorf, Universitätsstraße 1, 40225, Düsseldorf, Germany. till.freudenberger@uni-duesseldorf.de.
authors.forename: T
authors.initials: T
authors.lastname: Freudenberger
mesh_terms: D000818:Animals; D002196:Capillaries / Q000254:growth & development*; D004195:Disease Models, Animal; D006614:Hindlimb / Q000098:blood supply* / Q000473:pathology; D007150:Immunohistochemistry; D007511:Ischemia / Q000473:pathology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence; D018482:Muscle, Skeletal / Q000098:blood supply* / Q000473:pathology; D018919:Neovascularization, Physiologic
keywords: Angiogenesis; Capillary density; Hind limb ischemia; Immunohistochemistry