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Fig. 1. Speci¢city of antibodies against XLK. Proteins of tail skin of tadpoles at stage 54 were separated by 2-D SDS^PAGE and subjected to CBB
staining (A) or Western blot analysis with antibodies against XLK (B). The proteins enclosed by a broken line in A are those reacted with the antibodies.
The speci¢city of the antibodies was also immunohistochemically examined on tissue sections prepared from back skin of tadpoles at stage 56 (C)
and stage 61 (D). The structure of tadpole back skin at stage 56 and 61 is presented in Fig. 6A1 and B1, respectively, wherein the skin sections were
stained with H p E. The epidermis at stage 61 showed a weak background signal. The location of the collagen lamella in C and D was determined by
observing the section in a bright ¢eld and is indicated by two broken lines. ap, apical cells ; sk, skein cells ; bs, basal cells ; cl, collagen lamella; bm,
basement membrane; ct-1, connective tissues beneath the collagen lamella; ct-2, connective tissues between the basement membrane and the collagen lamella;
gl, secretory glands. Bar in C,D =20 Wm. |
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Fig. 4. Expression of xlk, xak-a, and xak-b during spontaneous metamorphosis. RNAs were extracted from the body (A) and tail (B) of tadpoles at different
metamorphic stages indicated. Ad in A represents RNAs of the adult body skin. These RNAs were electrophoretically separated on agarose gels
and were processed for Northern blot analysis. Northern blots were also done for rpL8 mRNA to estimate the amount of RNAs loaded on the gels. |
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Fig. 6. In situ hybridization of mRNAs of xlk, xak-a, and xak-b for the skin of tadpoles and adults. Cross sections of the body skin were prepared
from tadpoles at stage 56 (A1^A4), stage 61 (B1^B4), and from adults (C1^C4). Histology of the skin was examined by HpE staining (A1,B1,C1). The
epidermis at stage 56 (A1) was composed of apical (ap), skein (sk), and basal cells (bs). The epidermis of stage 61 (B1) transformed to the adult epidermis
composed of granular (gr) and basal cells. The adult epidermis (C1) contained corni¢ed cells (c) in addition to granular and basal cells. The skin at
stage 56 (A1) contained the subepidermal connective tissue (ct-1), but did not develop the connective tissue (ct-2) between the basement membrane (bm)
and the collagen lamella (cl), but the skin at stage 61 (B1) and the adult (C1) formed it. In the HpE section of adults the collagen lamella is out of vision.
The sections were hybridized with antisense probes of xlk (A2,B2), xak-a (A3,B3,C3), and xak-b (A4,B4,C4). The results with sense probes were
shown only for xak-a in adult tissues (C2), because all of the sense probes gave a similar background stain for all the tissues tested as in C2. f, Fibroplasts;
gl, secretory glands. Bar =20 Wm. |
