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Characterization of luteinizing hormone synthesized and released by rat pituitaries in vitro: dissociation of immunological and biological activities.

These studies were designed to characterize the LH synthesized de novo by rat anterior pituitary glands in vitro. Pituitaries from ovariectomized rats were incubated with [H]- glucosamine ([3H]GA), [14C]alanine, ([14C]A), and synthetic gonadotropin- releasing hormone for 4 h. At the end of incubation, pituitaries were separated from medium and then homogenized, centrifuged, and dialyzed. Dialyzed medium and supernatant were chromatographed separately on Sephadex G-100. Fractions were monitored for total immunoreactive LH (IR-LH) by RIA. Newly synthesized radiolabeled LH was measured by monitoring the radioactivity of LH immunoprecipitated with anti-LH-β serum. About 4% of the total IR-LH in the pituitary supernatant eluted in the void volume (peak A), whereas 96% eluted at the same position as [125I]rat LH (peak B). Seventy-six percent of total [14C]A-LH and 37% of total [3H]GA-LH eluted in peak A, whereas the remainder eluted in peak B. The ratio of [14C]A-LH to [3H]GA-LH in peak A varied from 2.5 to 3 5...