An immunochemical analysis of oxytocin and vasopressin prohormone processing in vivo.
Antisera against partially processed, unamidated forms of AVP and OT were raised and characterized by radioimmunoassay and immunocytochemistry. These antibodies, and antibodies that recognize fully processed, amidated forms of AVP and OT, were used together with various fractionation methods to study the content of prohormones, partially processed and fully processed forms of AVP and OT in the hypothalamo-neurohypophysial system of adult and fetal (E21) rats. The levels of cleaved AVP and OT in the fetus were lower than those of the adult (1 to 3 orders of magnitude for brain and pituitary, respectively), and the detection of cleaved OT in brain and pituitary was delayed compared to that of AVP. Pro-AVP cleavage efficiency in the adult and the fetus was high (99 and 95% cleavage, respectively) resulting in formation of fully processed amidated forms of AVP, with no detectable partially processed peptides. Pro-OT processing in the adult was very similar (over 99% cleavage) resulting in formation of fully processed amidated OT. However, Pro-OT processing efficiency in the fetus was very low and incomplete, resulting in 40% unprocessed precursor and the accumulation of C-terminally extended unamidated intermediate forms (OT-Gly, OT-Gly-Lys, and OT-Gly-Lys-Arg).
Alstein
M
M
Laboratory of Neurochemistry National Institute of Neurological and Communicative Disorders and Stroke, Bethesda, MD 20892.
Whitnall
M H
MH
House
S
S
Key
S
S
Gainer
H
H
D000818:Animals; D000906:Antibodies / Q000276:immunology; D001127:Arginine Vasopressin; D002850:Chromatography, Gel; D003429:Cross Reactions; D004586:Electrophoresis; D007030:Hypothalamo-Hypophyseal System / Q000276:immunology / Q000378:metabolism; D007150:Immunohistochemistry; D008297:Male; D009481:Neurophysins; D010121:Oxytocin / Q000031:analogs & derivatives* / Q000276:immunology / Q000378:metabolism; D011498:Protein Precursors / Q000276:immunology / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D011863:Radioimmunoassay; D051381:Rats; D011919:Rats, Inbred Strains; D013495:Supraoptic Nucleus / Q000276:immunology / Q000378:metabolism / Q000648:ultrastructure; D014667:Vasopressins / Q000276:immunology / Q000378:metabolism*
D000906:Antibodies; D009481:Neurophysins; D011498:Protein Precursors; D014667:Vasopressins; D001127:Arginine Vasopressin; D010121:Oxytocin; C046278:oxytocin, Gly-Lys-Arg-