title: Physiological concentrations of ouabain rapidly inhibit prolactin release from the tilapia pituitary.
abstract: Ouabain, a cardiac glycoside and inhibitor of Na(+), K(+)-ATPase, is now believed to be a steroid hormone in mammals. We have recently identified ouabain immunoreactivity in the plasma of the tilapia, a euryhaline teleost. Changes in plasma concentrations of immunoreactive ouabain (20-40 pM) in response to salinity change were well correlated with the changes in plasma osmolality and cortisol. Our previous studies have shown that cortisol rapidly inhibits prolactin (PRL) release from the tilapia pituitary by suppressing intracellular Ca(2+) ([Ca(2+)]i) and cAMP. In the present study, low doses of ouabain (10-1000 pM) inhibited PRL release dose-dependently during 2-24 h of incubation. There was no effect on growth hormone (GH) release, except for a significant increase at 1000 pM during 8-24 h of incubation. Significant dose-related increases in PRL release were observed at higher doses of ouabain (100-1000 nM), whereas significant inhibition was seen in GH release at 1000 nM during 2-24h of incubation. Ouabain at 1-100 pM had no effect on Na(+), K(+)-ATPase activity of the pituitary homogenate. The enzyme activity was inhibited by higher concentrations of ouabain, 10% at 1 nM, 15% at 10 nM, 28% at 100 nM, and 45% at 1000 nM. Ouabain also attenuated stimulation of PRL release by the Ca(2+) ionophore, A23187, and by a combination of dibutyryl cAMP and a phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthin. Intracellular Ca(2+) concentrations were monitored in the dispersed PRL cells with the Ca(2+)-sensitive dye, fura-2. Ouabain at 1 nM reversibly reduced [Ca(2+)]i within seconds, whereas 1 microM ouabain increased [Ca(2+)]i. A rapid reduction in [Ca(2+)]i was also observed when PRL cells were exposed to 1 microM cortisol, whereas there was no consistent effect at 1 nM. These results suggest that ouabain at physiological concentrations rapidly inhibits PRL release from the tilapia pituitary by suppressing intracellular Ca(2+) and cAMP metabolism. The stimulation of PRL release by high concentrations of ouabain (100-1000 nM) may result from an increase in [Ca(2+)]i, and subsequent depolarization due to the inhibition of Na(+), K(+)-ATPase activity.
authors.affiliation: Hawaii Institute of Marine Biology, University of Hawaii, Kaneohe, HI 96744, USA.
authors.forename: Shingo
authors.initials: S
authors.lastname: Kajimura
authors.forename: Andre P
authors.initials: AP
authors.lastname: Seale
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Hirano
authors.forename: Ian M
authors.initials: IM
authors.lastname: Cooke
authors.forename: E Gordon
authors.initials: EG
authors.lastname: Grau
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005260:Female; D010042:Ouabain / Q000494:pharmacology*; D010902:Pituitary Gland / Q000502:physiology; D011388:Prolactin / Q000378:metabolism*; D017210:Tilapia / Q000502:physiology*
chemical_list: D004791:Enzyme Inhibitors; D010042:Ouabain; D011388:Prolactin