Temporal-spatial co-localisation of tissue transglutaminase (Tgase2) and matrix metalloproteinase-9 (MMP-9) with SBA-positive micro-fibres in the embryonic kidney cortex.
Growth of the kidney is a complex process piloted by the collecting duct (CD) ampullae. The dichotomous arborisation and consecutive elongation of this tubular element determines the exact site and time for the induction of nephrons in the overlaying mesenchymal cap condensates. The mechanism by which the CD ampullae find the correct orientation is currently unknown. Recently, we have demonstrated micro-fibres that originate from the basal aspect of the CD ampullae and extend through the mesenchyme to the organ capsule. The micro-fibres are assumed to be involved in the growth and arborisation process of the CD ampulla. Therefore, we have investigated the specific distribution of the micro-fibres during branching morphogenesis. We have also analysed whether the micro-fibres co-localise with extracellular matrix (ECM)-modulating enzymes and whether the co-localisation pattern changes during CD ampulla arborisation. Micro-fibres were detected in all stages of CD ampulla arborisation. Tissue transglutaminase (Tgase2) co-localised with soybean agglutinin (SBA)-positive micro-fibres, whose presence depended upon the degree of CD branching. Matrix metalloproteinase-9 (MMP-9) also co-localised with micro-fibres, but its expression pattern was different from that for Tgase2. Western blotting experiments demonstrated that Tgase2 and MMP-9 co-migrated with SBA-labelled proteins. Thus, the micro-fibres are developmentally modulated by enzymes of the ECM in embryonic kidney cortex. These findings illustrate the importance of micro-fibres in directing CD ampulla growth.
Schumacher
Karl
K
Institute of Bioengineering and Nanotechnology, A*STAR, # 04-01, 31 Biopolis Way, The Nanos, 138669, Singapore. kmschumacher@ibn.a-star.edu.sg
Klar
Jürgen
J
Wagner
Charlotte
C
Minuth
Will W
WW
D000818:Animals; D000831:Animals, Newborn; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D005109:Extracellular Matrix / Q000201:enzymology; D019084:Fluorescent Antibody Technique, Indirect; D019204:GTP-Binding Proteins / Q000378:metabolism*; D007672:Kidney Cortex / Q000196:embryology / Q000201:enzymology*; D007685:Kidney Tubules, Collecting / Q000196:embryology / Q000201:enzymology*; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism*; D020894:Microfibrils / Q000201:enzymology*; D038081:Organogenesis; D037121:Plant Lectins / Q000378:metabolism*; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011817:Rabbits; D030262:Soybean Proteins / Q000378:metabolism*; D011503:Transglutaminases / Q000378:metabolism*; D010835:Phytohemagglutinins
D019204:GTP-Binding Proteins; D020780:Matrix Metalloproteinase 9; D037121:Plant Lectins; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D030262:Soybean Proteins; D011503:Transglutaminases; C028824:soybean lectin; D010835:Phytohemagglutinins