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  • Alternative splicing Kidney Laminins Whole-mount in situ hybridization Organ culture Mouse  (1)
  • Cell & Developmental Biology  (1)
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  • 1
    ISSN: 1432-0878
    Keywords: Alternative splicing Kidney Laminins Whole-mount in situ hybridization Organ culture Mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Laminins are large heterotrimeric basement membrane proteins that consist of α, β, and γ chains. We have previously shown that the human γ2 and γ2* transcripts result from the alternative use of the LAMC2 gene 3'-end exons. To explore the biological significance of the alternative γ2 transcripts, we isolated the cDNA coding for the mouse laminin γ2* transcript, characterized the 3'-end of the murine LAMC2 gene, and studied the expression of alternative γ2 transcripts in several mouse tissues. The sequence reported here is the first one containing a full-length γ2* 3'-UTR from any species. The mouse γ2* transcript is 4110 bases and encodes a putative polypeptide of 1110 amino acids. This polypeptide lacks the C-terminal cysteine residue thought to be important for heterotrimer formation. The mouse γ2* transcript was found to be expressed in several tissues by polymerase chain reaction (PCR), but at very low levels. The clearest signals were obtained on embryonic day 7, and in heart and testis of adult tissues. When the laminin γ2* transcript expression pattern was compared with that of the γ2 chain, a similar tissue distribution was observed. There was, however, a significant difference in expression levels. The longer γ2 transcript was found to be much more abundant than the shorter γ2* variant. Moreover, by whole-mount in situ hybridization, the shorter γ2* form was localized in the mesenchyme of the developing kidney whereas the longer γ2 form was exclusively present in the epithelium of the Wolffian (nephric) duct and ureteric bud. The results indicate different functions for the γ2 variants.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1058-8388
    Keywords: Type IV collagenases ; Tissue inhibitors of metalloproteinases (TIMPs) ; Trophoblast ; Embryo implantation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Expression of 72 kDa and 92 kDa type IV collagenases and the metalloproteinase inhibitors TIMPs 1, 2, and 3 was studied by in situ hybridization in implanting mouse embryos of days 5.5 to 7.5. The 92 kDa type IV collagenase was strongly expressed in invading trophoblasts, signals above background not being observed in the embryonic proper or placental tissue. In contrast, signals above background were not seen for the 72 kDa enzyme in any cells of the implantation region, including trophoblasts and stromal cells of the decidual tissue. Only cells in the mucosal stroma outside the decidual region displayed some expression. TIMP-3 was intensily expressed in maternal cells in the area surrounding the invading embryonic tissue. No expression was observed for TIMP-1 or TIMP-2 in the embryo proper, trophoblasts, or the area of the uterine decidual reactin. Weak signals appeared for TIMP-1 only in the circular layer of myometrial smooth muscle and in some uterine stroma cells distant from the site of embryo implantation. The results suggest a central role for 92 kDa type IV collagenase and TIMP-3 in the extracellular proteolysis associated with implantation of the early embryo. © 1995 Wiley-Liss, Inc.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
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