Neurohypophysial hormones as molecular evolutionary tracers: investigations on the sturgeons Acipenser stellatus and Acipenser guldenstadti

Rouillé, Yves; Chauvet, Jacqueline; Chauvet, M.T.; Acher, Roger; Polenov, A. L.

doi:10.1016/0305-0491(91)90280-q

Cited by 3 publications

(1 citation statement)

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“…The pressor activity of synthetic vasotocin is 250 milliunits/nmol (12), and from this value it is possible to calculate an amount of about 0. (22) and cyclostomes (23)(24)(25)(26)(27) (23)(24)(25)(26) and hagfishes (27,34). Because ofthe similar three-exon organization of mammalian oxytocin and vasopressin genes (14) and the preservation of the location of the three exons and the two introns in the hagfish vasotocin gene (34), it can be admitted that an early gene duplication occurred during evolution from cyclostomes to fishes-therefore, between 500 and 450 million years ago (28)-giving rise to two lineages of neurohypophysial peptides.…”

Section: Methodsmentioning

confidence: 99%

Special evolution of neurohypophysial hormones in cartilaginous fishes: asvatocin and phasvatocin, two oxytocin-like peptides isolated from the spotted dogfish (Scyliorhinus caniculus).

Chauvet

Rouillé

Chauveau

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

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In (6) and primitive (7) species. The question arises whether both duality and variability are proper to Selachii, in particular to sharks.We have now investigated a cartilaginous species, the spotted dogfish Scyliorhinus caniculus (order Pleurotremata, suborder Galeoidei) belonging to another suborder than Squalus acanthias (suborder Squaloidei). A duality of the oxytocin-like peptide has been discovered. Two new neurohypophysial hormones have been isolated and chemically and pharmacologically characterized. Vasotocin is also present in a considerably lower amount. MATERIALS AND METHODSPituitary Glands. Animals fished near the French Atlantic coast (in the vicinity of Lorient) were decapitated, and the heads in ice were sent to our laboratory so that pituitary glands were removed about 24 hr after death. Entire pituitaries were desiccated in peroxide-free and cool (40C) acetone, and then air-dried glands were powdered. The powder was titrated at 1.25 milliunits of oxytocic activity per mg on rat uterus (8). Pressor activity on anesthetized rat (9) was not measurable in crude extract. Two preparations of 1 g (about 200 glands) and 2.35 g (about 500 glands) were obtained by extraction with 0.1 M HCO for 4 hr at 40C (first preparation) and 0.25% acetic acid for 5 min at 1000C (second preparation) respectively. Supernatants were recovered by centrifugation.Molecular Sievings. Soluble material was freeze-dried either directly (first preparation) or after removal ofproteins by precipitating with 5% trichloroacetic acid (second preparation). Molecular sieving of preparation one was carried out on a column (2.1 x 31 cm) of Sephadex G-75 in 0.1 M formic acid to separate proteins. Both preparations were subjected to molecular sieving on a column (1.6 x 128 cm) of Bio-Gel P-4 in 0.1 M acetic acid. Three fractions from Bio-Gel P-4 (tubes 214-234, 235-250, 251-273 in the second preparation) containing oxytocic activity were pooled separately, concentrated, and subjected to HPLC.Reverse-Phase HPLC. Reverse-phase HPLC was used two or three successive times with a Nucleosil C18 column (4.5 X 210 mm; particle size, 5 gm) to obtain pure substances. The first elution system was a linear gradient of 5-50%o acetonitrile in 25 mM acetic acid (pH 3.0) for 40 min. The second system was a linear gradient of 10-25% acetonitrile in 12 mM sodium acetate (pH 5.0) for 40 min. The third system was used only for the second preparation and consisted of either a linear gradient of 10-15% acetonitrile in 21 mM acetate (pH 5) for 40 min for vasotocin or a linear gradient of 15-20%o acetonitrile in 4 mM acetate (pH 5) for 40 min for the two oxytocin-like hormones. 11266The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

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