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Estrogen Regulation of Peptidylglycine -Amidating Monooxygenase Expression in Anterior Pituitary Gland

INSERM U297, Institut Federatif de Recherche Jean Roche, Faculté de Médecine Nord, 13916 Marseille, Cedex 20, France

Address all correspondence and requests for reprints to: Dr. L’Houcine Ouafik, INSERM U297, IFR Jean Roche, Faculté de Médecine Nord, Bd Pierre Dramard, 13916 Marseille Cedex 20, France.

The pituitary is a rich source of peptidylglycine -amidating monooxygenase (PAM). This bifunctional protein contains peptidylglycine -hydroxylating monooxygenase (PHM) and peptidyl--hydroxyglycine -amidating lyase catalytic domains necessary for the two-step formation of -amidated peptides from their COOH-terminal glycine extended precursors. Expression of PAM was evaluated in the anterior pituitary of intact cycling adult female rat and after experimental manipulation of estrogen status. PAM messenger RNA (mRNA) levels showed changes inversely related to the physiological variations of plasma estrogen levels during the estrous cycle. Chronic treatment of ovariectomized (OVX) rats with 17 ß-estradiol decreased PAM mRNA levels to values comparable with those found in intact rats at proestrus. In situ hybridization of anterior pituitary sections using ³⁵S-labeled full length RNA antisense transcripts of rat PAM-1 complementary DNA showed that 17 ß-estradiol treatment induced an overall decrease of the hybridization signal, as compared with OVX rats. Progesterone treatment did not change PAM mRNA levels both in OVX or OVX + E₂ rats. Based on Northern blot analysis and amplification of fragments derived from rat PAM-1 by RT-PCR, it was found that estrogen status does not affect the distribution of PAM mRNA among its various alternatively spliced forms. In OVX 17 ß-estradiol treated rats, the specific activity of PAM in the anterior pituitary decreased in both soluble and particulate fractions compared with OVX animals. Western blot analysis demonstrated a 105-kDa PAM protein in particulate fractions prepared from OVX and OVX-17 ß-estradiol treated animals. The soluble fraction from OVX animals contained major PAM proteins of 105, 95, 84, 75, and 45 kDa, and 17 ß-estradiol treatment caused a decrease in the prevalence of these proteins. These results indicate that estrogens are involved, either directly or indirectly, in regulating the expression of PAM in several cell types in the anterior pituitary gland.

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