Expression of prostaglandin G/H synthase-1 and -2 in ovine amnion and placenta following glucocorticoid-induced labour onset.
Parturition in the sheep is preceded by an increase in the synthesis of prostaglandins by intrauterine tissues. Prostaglandin G/H synthase (PGHS) is the central enzyme involved in prostanoid production. Its expression is enhanced during late gestation in the ewe. Recent studies have identified two PGHS isozymes, termed PGHS-1 and PGHS-2. The labour-associated expression of the two isozymes of PGHS in the sheep has not been characterized. This study investigated the changes in expression of immunoreactive PGHS-1 and PGHS-2 in ovine amnion and placenta following glucocorticoid-induced labour. Ewes underwent surgery to implant fetal and maternal vascular cannulae and uterine electromyogram electrodes between 118 and 125 days of gestation. Fetal sheep were administered either the glucocorticoid betamethasone (n = 5) or saline (control n = 6) by direct transabdominal intrafetal injection. Ewes from the betamethasone-injected group were killed in the first stage of labour as indicated by uterine electromyographic activity. Ewes from the saline-injected group were killed at the same time to obtain age-matched control tissue. The time taken to euthanasia following induced-labour onset in the glucocorticoid-injected animals was 56.6 +/- 0.8 h post-injection. Plasma endocrine profiles in the maternal and fetal circulation following glucocorticoid injection were comparable to those observed following normal spontaneous delivery. At post-mortem, amnion and cotyledons were collected in liquid N2 and stored at -70 degrees C. Solubilized tissue extracts were prepared and analysed by Western blots using polyclonal antibodies to PGHS-1 and PGHS-2 isozymes. Fetal amnion contained PGHS-1 isozyme at day 133 of gestation, as demonstrated in the saline-injected animals. Slightly higher PGHS-1 immunoreactivity was observed following induced-labour onset, although this did not reach statistical significance (P > 0.05). PGHS-2 enzyme was not detectable in amnion. PGHS-2 expression was also not induced following labour onset. In contrast, PGHS-2 demonstrated enhanced expression following glucocorticoid-induced labour in ovine cotyledon. This tissue contained PGHS-1 enzyme, but immunoreactive levels were minimal and demonstrated limited regulation at labour. These data suggest that the previously reported rise in placental PG production at term in the sheep is predominantly due to increased expression of the PGHS-2 isozyme. This suggests that PGHS-2 contributes to PG production at term labour in sheep or is induced by the mechanisms controlling ovine parturition. PGHS-1 isozyme is produced constitutively in ovine amnion and may contribute to the gestational increase in PG formation by intrauterine tissues.
McLaren
W J
WJ
Department of Physiology, Monash University, Clayton, Victoria, Australia.
Young
I R
IR
Wong
M H
MH
Rice
G E
GE
D000650:Amnion / Q000737:chemistry / Q000201:enzymology*; D000818:Animals; D001623:Betamethasone / Q000494:pharmacology; D015153:Blotting, Western; D015237:Dinoprost / Q000031:analogs & derivatives / Q000097:blood; D015232:Dinoprostone / Q000097:blood; D005260:Female; D005312:Fetal Blood / Q000737:chemistry; D005938:Glucocorticoids / Q000494:pharmacology; D006854:Hydrocortisone / Q000097:blood; D007527:Isoenzymes; D007743:Labor, Obstetric / Q000187:drug effects / Q000378:metabolism*; D010920:Placenta / Q000737:chemistry / Q000201:enzymology*; D011247:Pregnancy; D011374:Progesterone / Q000097:blood; D011451:Prostaglandin-Endoperoxide Synthases / Q000032:analysis*; D012756:Sheep / Q000378:metabolism*
D005938:Glucocorticoids; D007527:Isoenzymes; C008228:15-keto-13,14-dihydroprostaglandin F2alpha; D011374:Progesterone; D001623:Betamethasone; D015237:Dinoprost; D011451:Prostaglandin-Endoperoxide Synthases; D015232:Dinoprostone; D006854:Hydrocortisone