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Prolactin (PRL) Receptors Are Colocalized in Dopaminergic Neurons in Fetal Hypothalamic Cell Cultures: Effect of PRL on Tyrosine Hydroxylase Activity¹

Department of Physiology, Southern Illinois University School of Medicine (L.A.A.), Carbondale, Illinois 62901-6512; and the Department of Physiology, University of Kansas Medical Center (J.L.V.), Kansas City, Kansas 66160-7401

Address all correspondence and requests for reprints to: Dr. Lydia A. Arbogast, Department of Physiology, Southern Illinois University School of Medicine, Carbondale, Illinois 62901-6512.

This study examined the responsiveness of dopaminergic neurons to PRL and the expression of PRL receptors in fetal hypothalamic cells. Hypothalamic cells were cultured in medium containing 5 or 25 mM potassium (K⁺) with or without 5% FBS. Rat PRL (rPRL) treatment (10–1000 ng/ml) for 10 days increased tyrosine hydroxylase (TH) activity 1.6- to 1.8-fold in dopaminergic neurons cultured in serum-containing medium with 25 mM K⁺, but not in defined medium or any medium with 5 mM K⁺. The rPRL-induced increase in TH activity was observed at 10–1000 ng/ml after both 1 and 10 days of rPRL treatment, whereas 1 ng/ml was not effective. TH activity was not altered after 1–12 h of rPRL treatment (100 ng/ml), but was increased 1.4-fold after 1–3 days and 1.8-fold after 5–10 days. The colocalization of PRL receptors and TH was evaluated by double labeled immunocytochemistry. PRL receptor immunostaining was observed in most TH-immunoreactive cells cultured in either defined or serum-containing medium with or without 10 days of rPRL treatment (100 ng/ml). As assessed by reverse transcriptase-PCR, the long form, but not the short form, of the PRL receptor was expressed in the hypothalamic cells regardless of medium composition, similar to the expression pattern in adult mediobasal hypothalamus from ovariectomized rats. These data indicate that a factor present in FBS imparts PRL responsiveness to hypothalamic dopaminergic neurons in vitro. The effective PRL concentrations and the time course for PRL’s action in vitro are within the physiological range in vivo. The colocalization of PRL receptor in dopaminergic neurons provides anatomical evidence for a direct effect of PRL, with the long form of the PRL receptor being the predominant form in the hypothalamic cells.

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