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Gen Comp Endocrinol
1996 Apr 01;1021:1-10. doi: 10.1006/gcen.1996.0039.
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Occurrence of immunoreactive activin/inhibin beta(B) in gonadotrophs, thyrotrophs, and somatotrophs of the Xenopus pituitary.
Uchiyama H
,
Komazaki S
,
Asashima M
,
Kikuyama S
.
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An antibody against the Xenopus activin/inhibin beta(B) subunit (94-107) was raised in a rabbit. Using this antibody, the distribution of activin/inhibin beta(B) immunoreactivity in the pituitary of adult X. laevis was studied. Beta(B) immunoreactivity was detected in gonadotrophs, thyrotrophs, and somatotrophs under light microscopy. Electron microscopy revealed that a beta(B)-immunoreactive substance exists in LH, TSH, and GH granules, in contrast to findings in the rat and goldfish. These results indicate that the expression of activin/inhibin beta(B) in pituitary cells is not consistent among vertebrate species.
FIG. 1. ELISA showing the specificity of anti-activin/inhibin bB
chain antibody. (A) Anti-bB chain antibody was applied to human
recombinant activin B-coated (solid circles) or human recombinant
activin A-coated (open squares) ELISA plates in a twofold dilution
series as shown by the log scale of the abscissa. (B) Anti-bB antibody
(10 mg/ml) was added to increasing concentrations of Xenopus bB
peptide (solid circles) or Xenopus bA peptide (open squares) as
shown by the log scale of the abscissa. These solutions were then
applied to bB peptide-coated ELISA plates. Each square or circle
represents the mean of three determinations.
FIG. 2. Immunohistochemical staining of Xenopus pituitary gland with various antibodies against pituitary hormones. The sections are
midsagittal paraffin sections from the same sample. Anterior is to the left and dorsal is to the top. (A)–(F) are of the same magnification. Bar:
200 mm. (A) Stained with anti-bB antibody; (B) stained with anti-bB antibody preabsorbed with Xenopus bB peptide; (C) stained with antibullfrog
LH b; (D) stained with anti-human TSH b; (E) stained with anti-bullfrog ACTH; (F) stained with anti-bullfrog GH. a, Anterior lobe;
i, intermediate lobe; p, posterior lobe.
FIG. 3. Enlarged view of an immunohistochemically stained Xenopus pituitary gland. (A)–(E) are paraffin sections from the same sample. The
signal is shown in brown, and nuclei are counterstained blue. (A) Whole pituitary view stained with anti-bB antibody. (Bar: 100 mm). In (B)–(G)
the sections shown at the top (B, D, and F) were prepared from sections adjacent to those in the bottom (C, E, and G). [(B–E) Bar: 30 mm. (F
and G) Bar: 15 mm.] Where the same cell body appears in adjacent sections, it is indicated by numbered arrows. (B and D) Stained with anti-bB
antibody. (C) Stained with anti-bullfrog LH b. (E) Stained with anti-human TSH b. (F and G) Adjacent 1-mm-thick Lowicryl K4M sections. The
signal appears blue and nuclei are counterstained red. (F) Stained with anti-bB antibody. (G) Stained with anti-bullfrog GH. a, Anterior lobe;
i, intermediate lobe; p, posterior lobe.
FIG. 4. Electron microscopical immunostaining of Xenopus pituitary gland. Except for (B), (A)–(J) are of the same magnification. The diameter
of the colloidal gold particles is 15 nm unless otherwise indicated. (A) FSH b-immunoreactive granules. (B) LH b-immunoreactive granules,
costained with anti-bB chain antibody (5-nm gold). (Bar: 250 nm.) (C) TSH b-immunoreactive granules. (D) TSH-like granules stained with
anti-bB antibody. (E) ACTH-immunoreactive granules. (F) ACTH-like granules showing a negative reaction with anti-bB antibody. (G)
GH-immunoreactive granules. (H) GH-like granules stained with anti-bB antibody. (I) PRL-immunoreactive granules. (J) PRL-like granules
showing a negative reaction with anti-bB antibody. (Bar: 500 nm.)
