Levi G et al. (1991), EP-cadherin in muscles and epithelia of Xenopus...

XB-ART-24304

Development 1991 Dec 01;1134:1335-44.

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EP-cadherin in muscles and epithelia of Xenopus laevis embryos.

Levi G , Ginsberg D , Girault JM , Sabanay I , Thiery JP , Geiger B .

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EP-cadherin is a novel Xenopus Ca+2-dependent adhesion molecule, which shares comparable homology with mouse E- and P-cadherins (Ginsberg, De Simone and Geiger; 1991, Development 111, 315-325). We report here the patterns of expression of this molecule in Xenopus laevis embryos at different developmental stages ranging from cleavage to postmetamorphic. EP-cadherin is already expressed in the oocyte and egg and can then be detected in close association with the membrane of all blastomeres up to late blastula stages. Starting at late gastrula stages, the level of EP-cadherin expression increases sharply in non-neural ectodermal cells, in the somites and in the notochord; it persists in endodermal cells and decreases rapidly in all migratory cells. During neurulation the level of EP-cadherin expression declines gradually in the nervous system and is undetectable here throughout later development except in the optic nerve and in the neural part of the olfactory organ. This pattern continues during later development so that in the tailbud stage and up to metamorphosis the most prominent staining is detected in the epidermis and skeletal muscle. After metamorphosis, the molecule gradually disappears from the muscle tissue and the major site of expression remains the skin. EP-cadherin is invariably present in close association with the cell membrane. In the muscle it is associated with the sarcolemma at regions of myoblast-myoblast or myotube-myotube contact. In epidermal cells, EP-cadherin is usually coexpressed with E-cadherin. Yet, while E-cadherin staining is always restricted to the basolateral aspects of the cells, EP-cadherin is often distributed throughout the plasmalemma including the apical surface.

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Species referenced: Xenopus laevis
Genes referenced: cdh1 cdh3

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	Fig. 1. Characterization of anti-EP-cadherin antibodies. (A) Western blot analysis of extracts of CHO cells stably transfected with either EP-cadherin cDNA (lanes 1,3) or the neomycin-resistance vector only (lanes 2,4) and of A6 Xenopus epithelial cells (lanes 5,6); the blots were reacted either with the pan-cadherin serum R-156 (lanes 1,2,5) or with anti-EP-cadherin antiserum R-827 (lanes 3,4,6). (B) Western blot analysis of extracts from eggs (lane 1), stage 8 blastulas (lane 2) and stage 17 neurulas (lane 3) performed with R-827 antiserum. (C and D) Immunofluorescence staining of CHO cells stably transfected with either EP-cadherin cDNA (B) or the neomycin-resistance vector only (C). EP-cadherin immunostaining is evident at the region of contact of the epithelioid polygonal transfected cells.
	Fig. 2. Distribution of EP-cadherin during early embryogenesis. Immunoperoxidase labeling of early blastula with anti-EP-cadherin (R-827). Bar=20um.
	Fig. 3. Immunofluorescent labeling of section of Xenopus larva using anti-EP-cadherin R-827 antibodies. Sections from stage 22 (A-E) and stage 41 (F) embryos. The positively labeled structures include: the cement gland and underlying endoderm (A), the lateral epidermis and underlying mesoderm and endoderm (B), the somites (C), the endoderm (D) and the notochord (E and F). ce, cement gland; e, epidermis; en, endoderm; me, mesoderm; n, notochord; s, somite; ph, pharynx. Bar=15/xm (A,B,E); 25/an (C,D); 40um (F).
	Fig. 4. Distribution of anti-EP-cadherin immunoreactivity in the skin. (A,B) Double staining with R-827 anti-EP-cadherin antibody (A) and a monoclonal anti Xenopus E-cadherin (B) of larval skin (stage 41) showing distinct distribution of the two molecules. Premetamorphic skin (C) and forelimb of a stage 51 animal (D) stained with R-827. Note that only the newly formed epidermis of the limb is stained for EP-cadherin while the degenerating epidermis of the body (sk) is not. fl, forelimb; g, gland; ol, outerlayer of the epidermis; sk, degenerating skin of the body; si, sensory layer of the epidermis. Bar=30um.
	Fig. 5. Expression of EP-cadherin in Xenopus muscles. Single immunolabeling of sections of stage 34 (A), 45 (B) and 53 (C) with R-827 anti-EP-cadherin antibody. Sections were from either the tail (A,B) o r the head (C). Double immunofluorescence labeling of stage 53 animals for EP-cadherin (D,F) and E-cadherin (E,G). Areas shown in D and E are from the head region while F and G are from the tail. Immunoblots of dissected tail axial muscles (H) and tail epidermis (I) with R-8327 corresponding to specimens shown in F. c, cartilage; e, epidermis; i, iris; m, muscles; r, retina. Bar=10/im (A,B); 25/an (C); 50jan (D-G).
	Fig. 6. Vinculin immunolabeling (A) and electron microscopy analysis (B) of Xenopus tail muscles. The arrows indicate electron-dense close contacts between the sarcolemma of adjacent myotubes.
	Fig. 7. Distribution of anti-EP-cadherin in the nervous system and sensory organs. (A,C) Longitudinal (A) and transverse (C) sections through the optic nerve of stage 41 (A) and 51 (B) Xenopus. The optic nerve is positive after its exit from the lamina cribrosa. (B) Staining of the spinal cord of a stage 51 tadpole. No staining is observed in the central nervous system beside the meninges and the ependymal canal. (D) Auditory organ at stage 41; the crista externa and part of the sensorial epithelium are stained. (E) Olfactory organ stage 41; the nasal pit and the Jacobson's organ are stained, on, optic nerve; ep, ependymal canal; mg, meninges; r, retina; ce, crista externa; ev, ear vesicle; np, nasal pit; Jo, Jacobson's organ. Bar=40um.