crine cells of the anterior pituitary are controlled by the central nervous system through hormonal interactions and are not believed to receive direct synaptic connections from the brain. Studies suggest that some pituitary cells may be modulated by the neurotransmitter glutamate (5, 16). We investigated prolactin (PRL)-releasing cells of the anterior pituitary of a euryhaline fish, the tilapia (Oreochromis mossambicus), for the presence of possible glutamate receptors (GluRs). Fura-2 imaging addressed the ability of glutamate to increase intracellular calcium. We observed a dose-dependent increase in intracellular calcium with transient perfusion (1-2 min) of glutamate (10 nM to 1 mM) in two-thirds of imaged cells. This increase was attenuated by the ionotropic GluR antagonist kynurenic acid (0.5-1.0 mM). The increase was also blocked or attenuated by antagonists of L-type voltage-gated calcium channels. The GluR agonist ␣-amino-3-hydroxy-5-methylisoxazole propionic acid (AMPA; 100 M) produced intracellular calcium increases that were reversibly blocked by the selective AMPA antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX). In contrast, the selective agonist N-methyl-D-aspartate (NMDA; 100 M to 1 mM in magnesium-free solution with 10 M glycine) had no effect on intracellular calcium. Radioimmunoassays demonstrated that glutamate stimulated PRL release. CNQX but not the NMDA receptor antagonist 2-amino-5-phosphonovaleric acid blocked this release. Antibodies for mammalian AMPA-and NMDAtype GluR produced a similar punctate immunoreactivity in the periphery of PRL cells. However, the NMDA antibody recognized a protein of a different molecular mass in PRL cells compared with brain cells. These results clearly indicate the presence of GluRs on tilapia PRL cells that can stimulate PRL release.pituitary; endocrine; osmoregulation; calcium imaging; radioimmunoassay THE ANTERIOR PITUITARY, or adenohypophysis, is derived from nonneuronal epithelial tissue that fuses during embryonic development with the neural crest-derived neurohypophysis (posterior pituitary) (17). Cells of the adenohypophysis release a variety of peptide hormones involved in growth, stress, metabolism, and sexual differentiation and function. In mammals, a complex portal system of capillaries carries blood and signaling molecules between the neurohypophysis and adenohypophysis. In teleost fish, this system is limited to a thin wall of capillaries between the anterior and intermediate lobes. The portal system brings hormones and releasing factors to cells of the anterior pituitary from neurosecretory cells in the hypothalamus. In addition, it carries hormones released from pituitary cells to the circulation.The major excitatory neurotransmitter in the vertebrate central nervous system is the amino acid glutamate. Recent research suggests that many nonneuronal cells also have glutamate receptors (16). Although glutamatergic innervation of endocrine cells of the anterior pituitary has not been described, growing evidence suggests that glutamate ca...