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The application of immunogold histochemistry to the quantitative analyses of basement membranes /

The objective of the thesis was to examine the distribution, biogenesis and assembly of basement membrane components in situ. This goal was achieved by immunolocalizing laminin, collagen IV, heparan sulfate proteoglycan and other components using 5 and 15nm colloidal gold particles. / The distribution of these components, showed them to be ubiquitously localized throughout the thickness of all basement membranes examined. In "thick" basement membranes: Reichert's membrane and the matrix of the mouse EHS tumor, and in the "thin" common basement membranes, the components were localized to the cord network (Inoue et al., 1985). Despite some variations between tissues, quantitation of gold particles indicated a tendency for the molar ratio of laminin:collagen:proteoglycan to be 1:1:0.2 in the thick and 1:1:1 in the thin basement membranes. / The biogenesis of laminin and collagen IV was examined in the endodermal cells producing Reichert's membrane. Both were co-localized in the rough endoplasmic reticulum, Golgi apparatus and secretory granules, corresponding to the pathway followed during processing. / Since the components are believed to self-aggregate after secretion, the molar ratios were used as guidelines to combine the three molecules in vitro, at 35$ sp circ$C. A precipitate formed, which in the electron microscope showed three main structures, one of these had the thickness, immunohistochemical and fine, ultrastructural characteristics of authentic basement membrane. Hence, basement membrane was reconstructed in vitro.

Identiferoai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.75748
Date January 1987
CreatorsGrant, Derrick Shawn
PublisherMcGill University
Source SetsLibrary and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada
LanguageEnglish
Detected LanguageEnglish
TypeElectronic Thesis or Dissertation
Formatapplication/pdf
CoverageDoctor of Philosophy (Department of Anatomy.)
RightsAll items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated.
Relationalephsysno: 000570071, proquestno: AAINL46098, Theses scanned by UMI/ProQuest.

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