Ontogenetic changes in the extra-pituitary expression of pro-opiomelanocortin in the developing ovine fetus.
Previous studies performed in this laboratory have demonstrated that the fetal lung contains immunoreactive adrenocorticotropin (irACTH), and that the lung both clears and secretes irACTH under basal and stimulated conditions. Furthermore, we have demonstrated that the irACTH in fetal lung is accounted for by proopiomelanocortin (POMC), and that there is an evidence of post-translational processing that is distinct from the pattern of processing typical of the anterior pituitary. The present study was designed to test the hypothesis that POMC is synthesized in the fetal lung, and that there is decreased synthesis in the late-gestation ovine fetal lung. Lungs were collected from fetal sheep at 80, 100, 120, 130, and 145 days gestation (n=4/group; term=147 days). POMC mRNA was measured using reverse transcription and real-time polymerase chain reaction with probe and primers designed in this laboratory. The greatest abundance of POMC mRNA was in the 80-days fetal sheep, and the relative abundance decreased as a function of fetal gestational age. POMC protein was measured using immunoblot analysis in lungs from 80, 120, and 145-days fetal sheep. The pattern of POMC protein abundance was consistent with that of the mRNA (highest at 80 days, lowest at 145 days). The POMC immunoblot revealed specific staining of a peptide with molecular weight of 27 kDa and another peptide with a molecular weight slightly higher than that of native POMC (32 kDa). For comparison, we measured POMC mRNA in skeletal muscle and small intestine. We found POMC expression in both fetal tissues, but no statistically significant ontogenetic pattern of expression. We conclude that POMC is synthesized in the ovine fetal lung, and that the rate of synthesis decreases as the fetus matures in utero. We speculate that the decreasing abundance of POMC mRNA and protein reflects decreased release of POMC and POMC-related peptides into the fetal bloodstream.
Ali
Nyima S
NS
Department of Physiology and Functional Genomics, University of Florida College of Medicine, P.O. Box 100274, Gainesville, FL 32610-0274, USA.
Keller-Wood
Maureen
M
Wood
Charles E
CE
D000818:Animals; D005865:Gestational Age; D015151:Immunoblotting; D007421:Intestine, Small / Q000737:chemistry; D008168:Lung / Q000378:metabolism*; D008970:Molecular Weight; D018482:Muscle, Skeletal / Q000737:chemistry; D010903:Pituitary Gland, Anterior / Q000196:embryology / Q000254:growth & development* / Q000378:metabolism*; D011333:Pro-Opiomelanocortin / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012756:Sheep
D012333:RNA, Messenger; D011333:Pro-Opiomelanocortin