Vethamany-Globus S (1989), Immunohistochemical localization of beta-endorp...

XB-ART-26613

Gen Comp Endocrinol 1989 Aug 01;752:271-9. doi: 10.1016/0016-6480(89)90080-4.

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Immunohistochemical localization of beta-endorphin-like material in the urodele and anuran amphibian tissues.

Vethamany-Globus S .

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In the present study, we have localized for the first time beta-endorphin (beta-EP)-like material in the adult and larval urodele and anuran tissues using immunohistochemical techniques. In the adult Notophthalmus viridescens and Ambystoma mexicanum, strong immunoreactivity to beta-EP antisera was observed in the region of the intermediate lobe, the latter fluorescing as a discrete body. The fluorescence was confined to the periphery of the cells, while the nuclei and the immediately surrounding cytoplasmic regions of the cells remained unstained. A few scattered cells in the anterior pituitary gland as well as the tracts in the posterior lobe also exhibited positive staining, although not as strong as the intermediate lobe. In the larval urodele, A. maculatum, beta-EP-like material was localized for the first time in the sensory ganglia and their emerging nerve fibers, in the Leydig cells of the skin, as well as in a few discrete cells scattered among stomach epithelial cells. In addition to the above, immunoreactivity to beta-EP antisera was observed in the cellular intermediate part of the neurointermediate lobe of the pituitary gland in young Xenopus laevis, the neural part of the lobe remaining nonreactive.

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Species referenced: Xenopus laevis
Genes referenced: pomc

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	FIG. la AND lb. Sag&al sections through the intermediate lobe of newt pituitary showing bright fluorescence to g-EP antibody. The staining is confined to the periphery of the cells of pars intermedia, while the nuclei and the central part of the cells remained unstained. Specificity was continned by abolishing fluorescence, using antibody which was preincubated with B-EP antigen (le). x310; X 1200, respectively; c, cell; pd, pars distalis; pi, pars intermedia; pn, pars nervosa. FIG. lc AND Id. Sagittal sections through an axolotl pituitary showing immunofluorescence to B-EP antibody in the intermediate lobe. The pattern of fluorescence is very similar to that observed in the newt pituitary. X310; X1200, respectively. FIG. le AND If. Absorption controls of newt pituitary sections. (e) Total abolishing of fluorescence by treatment with B-EP antiserum which was preincubated with 8-EP antigen. The remaining unblocked fluorescence in (f) may represent cross-reactivity of the antiserum to B-lipotropin. x310; 1200, respectively.
	FIG. 2a-2d. Sagittal sections of Larval A. maculatum; (a, b) the spinal ganglia (g) adjacent to the vertebral cartilage (c) exhibiting strong immunoreactivity. FIG. 2c AND 2d. B-EP antisera show immunofluorescence to B-EP antisera localized not only in the perikarya (d) of the larval sensory neurons, but also along the periphery of the emerging nerve fibers (c). n, nucleus; p, periphery of cell body; ne, nerve fibre. Fig. 2a-2c x200; d X9(@. FIG. 2e. Bright fluorescence to B-EP antisera is localized in the Leydig cells (L) of the larval skin of A. maculatum. ~225. FIG. 2f. Localization of B-EP immunoreactivity in the cells (c), scattered among lining (ep) of the stomach of larval A. maculatum. s, lumen showing stomach contents; x225.
	FIG. 3a. A frontal section through xenopus pituitary, stained with hematoxylin and eosin. The neurointermediate lobe is bilobed and each lobe is situated on either side of pars distalis (pd), consisting of the cellular intermediate lobe (pi) and the neural lobe (pn) made predominantly of nerve fibers. x80. FIG. 3b. Immunocytochemical localization of g-EP in the cellular part (pi) of the neurointermediate lobe; the neural part (pn) remains unstained. Immunofluorescence is confined to the cytoplasm of the cells, with the nuclei (n) remaining nonreactive to g-EP antiserum. x250. FIG. 3c AND 3d. Cells of the intermediate lobe of xenopus pituitary showing strong immunoreactivity to l3-EP antiserum. Much of this staining was abolished as seen in the section (d) which was stained with g-EP antiserum preincubated with p-EP antigen. The remaining unblocked staining may represent cross-reactivity of the antiserum to g-lipotropin. x 1000.