XB-ART-19219Dev Biol 1995 Oct 01;1712:531-40. doi: 10.1006/dbio.1995.1302.
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Development of the Xenopus pronephric system.
The pronephros serves as the embryonic kidney of the lower vertebrates. In this report we describe the development of the pronephric system of Xenopus laevis utilizing scanning electron microscopy and novel monoclonal antibodies that specifically recognize different parts of the pronephros. Antibody 3G8 recognizes the tubules and nephrostomes of the pronephroi only and does not react with the duct. Antibody 4A6 stains only the duct and the nephrostomes. These antibodies thus allow the positive identification of these two intermediate mesoderm derivatives. Both reagents detect antigens expressed some time after the pronephric structures first form and probably represent markers of terminal differentiation. When the tubules and duct first form they are separate structures that can easily be distinguished; the connective tubules have a distinctive organization, the collecting (or common) tubule is broader than other tubules, and the narrow pronephric duct has a specific shape and position. In later stages the collecting tubule and the rostral portion of the duct undergo a considerable amount of convolution, and both contribute to the final coiled tubular body of the pronephros. The ability of 3G8 and 4A6 to distinguish these two elements of the nephric system was used to reexplore classical experiments on the interaction between these two structures during development of the pronephric system. The use of whole-mount analysis has allowed us to examine large numbers of embryos from different stages and dissected in a variety of planes. These experiments demonstrate the dynamic nature of the intermediate mesoderm and indicate that although the pronephros may be specified by mid-neurula stages, patterning is not complete until tailbud stages.
PubMed ID: 7556934
Article link: Dev Biol
Antibodies: Kidney Ab1 Kidney Ab2
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|Fig 1. Characterization of monoclonal antibodies. (A) Transverse section through a pronephric tubule stained with antibody 3G8 and developed with a blue substrate. Staining is localized to the apical surface of the tubule. (B) Transverse section through the pronephric duct stained with antibody 4A6 and developed with a blue substrate; staining is intense around the entire cell surface of the pronephric duct epithelia. The arrow indicates the layer of melanocytes lining the gut, which are distinguishable from antobody staining using phase contrast (not shown). (C) Western blot of stage 47 pronephric extract developed with antibody 4A6. A single band of 50 x 10  Mr is detected.|
|Fig. 2. Formation of the Xenopus pronephric tubules. Immunostaining with antibody 3G8 compared to scanning electron microscopy.|
|Fig. 3. Antibody 4A6 recognizes the pronephric duct|
|Double stained embryo. 3G8 staining appears dark blue. The red stain is a second antibody, 4A6|
|A. 3G8 stained in blue, 4A6 stained in red. B. 4A6 C. 4A6 D. 4A6|
|panel B, 4A6 staining the membrane or cortex of all pronephric duct cells|