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Mammary cells were prepared from lactating mice by collagenase digestion in order to examine the reaction of prolactin (PRL) with its cellular receptors. Cells were treated with 10% acetic acid following PRL binding. Between 5 and 12.5% acetic acid, the dissociation of PRL remained constant. The ratio of dissociated PRL was comparable to that of tissue slices or disrupted cells with this treatment. In subcellular membranes, about 81% of PRL bound to plasma membrane receptors was dissociated by acid treatment while PRL dissociated from intracellular membrane receptors was about 21%. Based on the nature of their PRL dissociation, PRL receptors were classified as acid-sensitive or acid-insensitive. The reaction of PRL with either species was reversible and saturable, and was dependent on temperature and time. Both species showed high PRL-binding affinity. However, acid-insensitive receptor was unstable at 37 degrees C. The level of acid-sensitive receptor was close to that of acid-insensitive receptor in late pregnancy. During the first 3 days of lactation, the level of acid-sensitive receptor increased more slowly than that corresponding to acid-insensitive receptor. The above criteria suggest that the postpartum increase in PRL binding is characterized especially by an increase in the level of acid-insensitive PRL receptor located mainly on intracellular membrane.

Characterization of Plasma and Intracellular Membrane Prolactin Receptor in Lactating Mouse Mammary Cells.