Purification of a Non-dopaminergic and Non-GABAergic Prolactin Release-Inhibiting Factor (PIF) in Sheep Stalk-Median Eminence

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Abstract
Prolactin release-inhibiting factor (PIF) extracted from 1200 sheep stalk-median eminences was purified by gel filtration on a Sephadex G-25 column (4.5 .times. 150 cm). PIF activity was determined by measuring the inhibition of prolactin release from dispersed anterior pituitary cells of adult male or estrogen-primed, ovariectomized rats. Using this system, PIF was detected in tube fractions 122-127 (volume = 20 ml/tube). These fractions also contained LHRH and somatostatin; these peptides had no prolactin-inhibiting activity in the quantities present. No dopamine or GABA detected in the active fractions by radioenzymatic assay and fluorophotoenzymatic assay, respectively. Receptor blockers for dopamine or GABA did not interfere with the PIF activity. The PIF activity cannot be attributed to either dopamine or GABA, both of which are known to inhibit prolactin release and provide evidence for the presence of a non-dopaminergic and non-GABAergic PIF within the hypothalamus.