title: Bioaccumulation of ergovaline in bovine lateral saphenous veins in vitro.
abstract: Ergot alkaloids have been associated with vasoconstriction in grazing livestock affected by the fescue toxicosis syndrome. Previous in vitro investigations studying how ergot alkaloids caused vasoconstriction have shown that ergovaline has a distinct receptor affinity and sustained contractile response. A similar contractile response has not been noted for lysergic acid. The objectives of this study were to determine if repetitive in vitro exposure of bovine lateral saphenous vein to lysergic acid or ergovaline would result in an increasing contractile response and if a measurable bioaccumulation of the alkaloids in the vascular tissue occurs over time. Segments of vein were surgically biopsied from healthy, Angus x Brangus cross-bred, fescue-naïve yearling heifers (n = 16) or collected from healthy mixed breed and sex cattle immediately after slaughter (n = 12) at a local abattoir. Veins were trimmed of excess fat and connective tissue, sliced into cross-sections, and suspended in a myograph chamber containing 5 mL of oxygenated Krebs-Henseleit buffer (95% O(2)/5% CO(2); pH = 7.4; 37 degrees C). Contractile responses to repetitive additions of ergovaline (1 x 10(-9) and 1 x 10(-7) M) and lysergic acid (1 x 10(-5) and 1 x 10(-4) M) were evaluated using the biopsied veins. For the bioaccumulation experiments, veins collected at the abattoir underwent repetitive additions of 1 x 10(-7) M ergovaline and 1 x 10(-5) M lysergic acid and the segments were removed after every 2 additions and media rinses for alkaloid quantification via HPLC/mass spectrometry. Contractile data were normalized as a percentage of contractile response induced by a reference dose of norepinephrine (1 x 10(-4) M). Repetitive additions of 1 x 10(-9) M ergovaline and 1 x 10(-5) and 1 x 10(-4) M lysergic acid resulted in contractile response with a negative slope (P < 0.02). In contrast, repetitive addition of 1 x 10(-7) M ergovaline resulted in a contractile response that increased with each addition (P < 0.01). Lysergic acid and ergovaline were detected at all 4 exposure levels (2x to 8x), but only the 1 x 10(-7) M ergovaline treatment resulted in increased tissue content as the number of exposures increased (P < 0.05). These data indicate that ergovaline, but not lysergic acid, bioaccumulates with repetitive exposure in vitro. These results suggest that ergovaline may have a greater potential for inducing toxicosis in grazing animals than lysergic acid because of its potential to bioaccumulate at the cellular site of action.
authors.affiliation: Forage-Animal Production Research Unit, U.S. Department of Agriculture-Agricultural Research Service, Lexington, KY 40546, USA. james.klotz@ars.usda.gov
authors.forename: J L
authors.initials: JL
authors.lastname: Klotz
authors.forename: B H
authors.initials: BH
authors.lastname: Kirch
authors.forename: G E
authors.initials: GE
authors.lastname: Aiken
authors.forename: L P
authors.initials: LP
authors.lastname: Bush
authors.forename: J R
authors.initials: JR
authors.lastname: Strickland
mesh_terms: D000818:Animals; D002417:Cattle; D004879:Ergotamines / Q000378:metabolism* / Q000494:pharmacology; D005260:Female; D008237:Lysergic Acid / Q000378:metabolism; D008297:Male; D012501:Saphenous Vein / Q000378:metabolism*; D014661:Vasoconstriction / Q000187:drug effects; D014662:Vasoconstrictor Agents / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D004879:Ergotamines; D014662:Vasoconstrictor Agents; C076760:ergovaline; D008237:Lysergic Acid