Differential biosynthesis and posttranslational processing of vasopressin and oxytocin in rat brain during embryonic and postnatal development

Abstract

The biosynthesis and posttranslational processing of arginine vasopressin (AVP) and oxytocin (OT) peptides in the developing rat brain and pituitary were studied using antibodies and complementary separation methods that permitted a quantitative radioimmunoassay (RIA) analysis of precursor, intermediate, and completely processed forms of the peptides. Precursor forms of the peptides were first detected in rat brain as early as embryonic day (E) 15 for AVP and E17 for OT. Proteolytic cleavage products of the precursors were detected 1 d later for both peptides. AVP was present in a fully processed (amidated) from immediately (E16) and throughout fetal development. OT was cleaved from its precursor starting on E18 but remained in an intermediate (C-terminal extended) form until E21, when amidated OT was first detected in the pituitary. Hence, Pro-AVP processing in the fetus was immediate and complete, whereas Pro-OT processing in the fetus was much slower and incomplete, resulting in the generation of partially processed, nonamidated stable forms of the peptide (OT-Gly10, OT-Gly10-Lys11, and OT-Gly10-Lys11-Arg12). The presence of OT-Gly10-Lys11-Arg12 as a major, stable intermediate form, indicated that the in vivo pattern of endoproteolytic cleavage occurred principally at the C-terminus of the pair of basic amino acids at the tripeptide spacer sequence (Gly-Lys-Arg) in the precursor. Although both precursors were first expressed nearly simultaneously in the brain, the steady-state levels of the precursors were very different throughout fetal life. From E16-E21, the quantities of AVP precursors and peptides were 5- to 10-fold greater than those of OT, suggesting a much higher level of precursor biosynthesis in the AVP neurons. In addition to these differences in the regulation of biosynthesis and processing, AVP peptides were axonally transported to the pituitary 3 d earlier than OT peptides, and in far greater (20-fold) abundance. The early presence and abundance of amidated AVP in the brain and pituitary suggests a trophic function for this peptide during development.