Ascorbic acid potentiates the inhibitory effect of dopamine on prolactin release in primary cultured rat pituitary cells

Abstract

ABSTRACT The chemical structure of dopamine includes an ortho-catechol group which is labile to oxidizing agents. Ascorbic acid, a reducing agent, has in the past been added to the incubation medium in order to protect dopamine against oxidation. However, there has been no thorough examination of the biological effect of ascorbic acid on prolactin release. In this present study we have shown that ascorbic acid has neither a stimulatory nor an inhibitory effect on prolactin release but reduces by approximately two orders of magnitude the concentration of dopamine necessary to inhibit prolactin release from cultured anterior pituitary cells. The strong potentiation effect of ascorbic acid was reproduced using apomorphine. We compared the effect of ascorbic acid and isoascorbic acid on dopamine inhibition of prolactin release. Isoascorbic acid is an epimer of ascorbic acid, having the same reduction–oxidation potential as ascorbic acid, but is less biologically active. Isoascorbic acid was less effective in potentiating the dopaminergic effect than was ascorbic acid, which supports the notion that potentiation by ascorbic acid is not entirely due to its reducing property. In order to dissociate further the chemical protection of dopamine from the biological potentiation, the inhibitory effects of freshly made and 3-h-old dopamine solutions were compared. Neither one of the two solutions contained any ascorbic acid, yet the two solutions did not show any difference in their ability to inhibit prolactin release during the 3-h incubation period, indicating that no significant amount of dopamine was oxidized. The minimum effective concentration of ascorbic acid necessary to demonstrate potentiation was between 0·001 and 0·01 mmol/l. The potentiation effect was shown after 1, 2, or 3 h of exposure to dopamine, and was evident in both 2- and 6-day-old cultured cells. The effect of ascorbic acid can either be a pharmacological potentiation or a physiological effect on the primary cultured pituitary cells. However, it is quite clear that ascorbic acid is not a simple anti-oxidant but produces a strong potentiating effect on the dopaminergic inhibition of prolactin release by some other means. J. Endocr. (1988) 118, 287–294