Somatostatin biosynthesis by cerebral cortical cells in monolayer culture.

Abstract

We examined the ability of isolated rat cerebral cortical cells to synthesize and secrete immunoreactive somatostatin (IRS). Dispersed telencephalic cells (10(7] from rat embryos contain approximately 24 pg IRS. The IRS content of 10(7) cultured cells increased steadily to a level of 2800 pg by 20 days in vitro. The IRS content in the medium followed a parallel production curve and was, in general, 10% of the cellular content. The nature of this IRS was characterized by gel filtration. Bio-Gel P-10 chromatography resolved four discrete IRS peaks. The first peak had an apparent molecular weight of 11,500. The second peak, which comigrated with synthetic Somatostatin-28, had a molecular weight of 7,000. The third peak had a molecular weight of 1,800, and the fourth peak, which comigrated with Somatostatin-14, had a molecular weight less than 1,800. Incubation with [3H]phenylalanine (Phe) revealed that 3H was incorporated into each peak. Isolation of 3H-labeled peak 4 was carried out by double affinity column purification. Sequential automated Edman degradation of this material revealed [3H] Phe in the positions that Phe occupies in Somatostatin-14. These studies document the ability of isolated rat cerebral cortical tissue to synthesize several forms of IRS, establishing them as endogenous cortical peptides.