Characterization of monoclonal antibodies against tenascin-C: no apparent effect on kidney development in vitro.
Tenascin-C is an extracellular matrix glycoprotein found in embryonic mesenchyme. The precise biological function of tenascin-C is unknown, but different parts of the molecule have effects on cell adhesion and other cellular activities. We studied the expression and role of tenascin-C in the embryonic mouse kidney. By Northern blots, no tenascin-C was detectable in uninduced mesenchyme from day 11 embryonic kidneys, but after 24 hours of in vitro culture both major splice variants of tenascin-C were detected. The larger variant was the predominant form. By in situ hybridization tenascin-C mRNA in 13-day old embryonic kidneys was detected in the mesenchyme surrounding newly formed epithelial structures. In 17-day old embryonic kidneys, tenascin-C mRNA was detected in mesenchyme around the forming epithelial structures in the cortex, and expression was also seen in mesenchyme surrounding the capsular epithelium of glomeruli. In newborn kidneys, expression had shifted to the medulla but was still confined to mesenchymal areas. We have characterized 6 new monoclonal antibodies against mouse tenascin-C, which all stain embryonic kidneys from different stages in a pattern consistent with earlier reports and with the mRNA data. The binding sites of the monoclonal antibodies on the tenascin-C molecule were mapped to discrete regions of tenascin-C. These six and five previously described antibodies against tenascin-C were tested in antibody perturbation experiments. Three of these have been shown by in vitro assays to perturb function of other cell types. Despite this, none of them inhibited development of mouse kidneys in organ culture, although they were tested at 1 mg/ml. It raises the possibility that tenascin-C is not crucial for kidney development. Alternatively, tenascin-C has more subtle functions which could not be identified with the assays used here.
Talts
J F
JF
Department of Animal Physiology, Uppsala University, Sweden.
Eng
H
H
Zhang
H Y
HY
Faissner
A
A
Ekblom
P
P
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D004627:Embryonic Induction; D004848:Epithelium / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D015870:Gene Expression; D017403:In Situ Hybridization; D007668:Kidney / Q000196:embryology* / Q000254:growth & development; D008648:Mesoderm / Q000378:metabolism; D051379:Mice; D018613:Microscopy, Confocal; D009924:Organ Culture Techniques; D012333:RNA, Messenger / Q000235:genetics; D019063:Tenascin / Q000276:immunology / Q000502:physiology*
D000911:Antibodies, Monoclonal; D012333:RNA, Messenger; D019063:Tenascin