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  • Direct immunogold histochemistry  (1)
  • Key words. Heart development; extracellular matrix; heart valves; hyaluronan; immunogold staining; microfibrils.  (1)
  • Laminin subunits of A- and B1-chains  (1)
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  • 1
    ISSN: 1432-0568
    Keywords: Mouse embryo day 6 and 7 ; Direct immunogold histochemistry ; Laminin subunits of A- and B1-chains ; Mesoderm formation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Using ultrastructural immunogold histochemistry on LR-Gold-embedded 6- and 7-day-old mouse embryos we investigated the appearance of the A- and B1-chains of the laminin molecule during mesoderm formation. With the help of antibodies against the A-chain and the E4 fragment of the B1-chain of the laminin molecule we were able to detect the subunits in vivo. Staining for the E4 fragment of the short arm of the laminin molecule from day 6 was negative. In contrast, strong staining for the A-chain of laminin was observed. Our results show, that the A-chain of laminin appears before the B1-chain in the 6-day-old mouse embryo before a basement membrane is seen between the ectodermal and entodermal cell layers. Furthermore, the staining pattern indicates, that the laminin molecule changes its orientation in the basement membrane of the ectoderm during mesoderm formation. On day 7 staining for the A-chain of laminin and for the E4 fragment was seen in a random distribution throughout the entire basement membrane, whereas in areas were the onset of mesoderm formation was taking place, the E4 fragment was restricted to the edge of the disintegrating basement membrane.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1420-9071
    Keywords: Key words. Heart development; extracellular matrix; heart valves; hyaluronan; immunogold staining; microfibrils.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The microfibrillar proteins fibulin-1 and fibulin-2 were previously identified as prominent components of the endocardial cushion tissue (ECT) during heart development and shown to persist in adult valves and septa. Immunogold staining has now been used to compare their localization in embryonic (days 9–11) and adult mouse heart with that of fibronectin and the chondroitin sulphate proteoglycan versican. All four proteins were deposited in the ECT, which consists of a hyaluronan-rich, mainly unstructured matrix, but were barely detectable in myocardial basement membranes or within endocardial cells. Digestion with hyaluronate lyase selectively released the fibulins and versican but not fibronectin from the ECT. Yet neither of the two fibulins bound to hyluronan in solid-phase assays, in contrast to versican. In the adult heart valve, all four proteins could be detected close to cross-striated collagen fibrils or microfibrils, but only versican was lost upon exposure to hyaluronate lyase. The data indicate that fibulins are associated with the hyaluronan-matrix of ECT through a bridge of versican, but that this association changes upon valve development to another supramolecular, presumably microfibrillar organization based on fibronectin and/or fibrillins.
    Type of Medium: Electronic Resource
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