Itoh K et al. (1988), The expression of epidermal antigens in Xenopus...

XB-ART-27334

Development 1988 Sep 01;1041:1-14.

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The expression of epidermal antigens in Xenopus laevis.

Itoh K , Yamashita A , Kubota HY .

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Five kinds of monoclonal antibodies that are specific for the epidermis of Xenopus embryos were produced. Epidermis-specific antibodies were used to investigate the spatial and temporal expressions of epidermal antigens during embryonic and larval development. The cells that were recognized by the antibodies at the larval stage are as follows: all of the outer epidermal cells and cement gland cells were recognized by the antibody termed XEPI-1, all of the outer and inner epidermal cells, except the cement gland cells, were recognized by XEPI-2 antibody, the large mucus granules and the apical side of the outer epidermal cells, except for the ciliated epidermal cells, were recognized by XEPI-3 antibody, the large mucus granules and basement membrane were recognized by XEPI-4 antibody, and the small mucus granules contained in the outer epidermal cells as well as extracellular matrices were recognized by the antibody termed XEPI-5. All of the epidermal antigens, except XEPI-4, were first detected in the epidermal region of the late gastrula or early neurula. The XEPI-4 antigen was first detected in stage-26 tail-bud embryos. None of these antigens were expressed by the neural tissues at any time during embryonic development. Only the XEPI-2 antigen continued to be expressed after metamorphosis, while the expression of the other antigens disappeared during or before metamorphosis. The specificity of the antibodies allowed us to classify the epidermal cells into four types in early epidermal development. The four types of epidermal cells are (1) the outer epidermal cells that contain small mucus granules, (2) the ciliated epidermal cells, (3) the outer epidermal cells that contain large mucus granules and (4) the inner sensorial cells.

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Species referenced: Xenopus laevis
Genes referenced: rpsa

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	Fig. 1. Expression of the XEPI-1 antigen in Xenopus embryos. Transverse polyester wax sections were stained by indirect immunofluorescence with XEPI-1 and FITC-conjugated rabbit antimouse IgG. (A) Stage 13; (B) stage 18; (C) stage 24; (D) stage 40. Bar, 100 um.
	Fig. 2. Regional distribution of antigens recognized by XEPI-2 in Xenopus embryos. Transverse polyester wax sections were stained with XEPI-2 and FITC-conjugated rabbit antimouse IgG. (A) Stage 16; (B) stage 20; (C) stage 26; (D,E) stage 36. Bar, 100um
	Fig. 3. Expression of the XEPI-3 antigen in Xenopus embryos. Transverse polyester wax sections were stained with XEPI-3 and FITC-conjugated rabbit antimouse IgG. (A) Stage 16; (B) stage 22; (C) stage 24; (D,E) stage 36. One of the ciliated epidermal cells is indicated by the arrowhead. Bar, 100um.
	Fig. 4. Regional distribution of antigens recognized by XEPI-4 in Xenopus embryos. Transverse polyester wax sections were stained with XEPI-4 and FITC-conjugated rabbit antimouse IgG. (A) Stage 28; (B,C) stage 36. Bar, 100um
	Fig. 5. Regional distribution of the antigens that were recognized by XEPI-5 antibody in Xenopus embryos. Transverse polyester wax sections (A-E) were stained with XEPI-5 and FFTC-conjugated rabbit antimouse IgG. (A) Stage 8; (B) stage 13; (C) stage 20; (D) stage 28; (E) stage 40. (F) Ultrastructural localization of the XEPI-5 antigen at stage 41 in a section of epidermal cells that were fixed with 2 % paraformaldehyde and embedded in LR gold. (A-E) Bar, 100urn. One of the ciliated epidermal cells is indicated by an arrowhead, bl, blastocoel. Bar, 5um
	Fig. 6. Comparison of distribution of antigens recognized by five antibodies at stage 36. Transverse polyester wax sections were stained with XEPI-1 (A), -2 (B), -3 (C), -4 (D). or -5 (E) and FITC-conjugated antimouse IgG. C and D were serial sections. Two of the ciliated epidermal cells are indicated by arrowheads. Bar, 100um.
	Fig. 7. Expression of the XEP1-1 antigen before and during metamorphosis. Transverse polyester wax sections were stained as described in Fig. 1. B-F were dorsal epidermal sections. (A) Stage 40; (B) stage 54; (C) stage 58; (D) stage 60; (E) stage 62; (F) stage 64. Bar, 100um.
	Fig. 8. Expression of the XEPI-2 antigen before, during and after metamorphosis. Transverse polyester wax sections were stained as described in Fig. 2. C-F were dorsal epidermal sections. (A) Stage 40; (B) stage 46; (C) stage 58; (D) stage 62; (E) stage 64; (F) stage 66. Bar, 100um.
	Fig. 9. Expression of the XEPI-3 antigen before and during metamorphosis. Transverse polyester wax sections were stained as described in Fig. 3. C-F were dorsal epidermal sections. (A) Stage 40; (B) stage 52; (C) stage 58; (D) stage 60; (E) stage 62; (F) stage 64. mu, muscles. Bar, 100um.
	Fig. 10. Regional distribution of the antigens recognized by the XEPI-4 antibody before and during metamorphosis. Transverse polyester wax sections were stained as described in Fig. 4. C-F were dorsal epidermal sections. (A) Stage 40; (B) stage 52; (C) stage 58; (D) stage 60; (E) stage 62; (F) stage 64. mu, muscles. Bar, 100um.
	Fig. II. Expression of the XEPI-5 antigen before and during metamorphosis. Transverse polyester wax sections were stained as described in Fig. 5. C-F were sections of dorsal epidermis. (A) Stage 40; (B) stage 46; (C) stage 58; (D) stage 60; (E) stage 62; (F) stage 64. Bar, 100um.
	Fig. 12. Proteins contained in the homogenate of stage- 35/-36 embryos were separated using SDS-PAGE on a 10% polyacrylamide gel (lane A). These proteins were blotted on a nitrocellulose sheet and incubated with the XEPI-1 antibody and 35S-labelled anti-mouse IgG (lane B). The XEPI-1 antibody recognized one major protein band having an estimated Mr of 250xl03 (lane B). Proteins contained in the homogenate of stage-54 epidermal tissues were separated using SDS-PAGE on a 4-20% polyacrylamide gradient gel (lane C). These proteins were blotted on a nitrocellulose sheet and reacted with the XEPI-2 antibody and 35S-labelled antimouse IgG (lane D). The XEPI-2 antibody recognized a total of six protein bands having estimated JW,S between 45 and 67X103 from epidermal proteins.