Vallet J L, Bazer F W, Fliss M F, Thatcher W W
Department of Animal Science, University of Florida, Gainesville 32611.
J Reprod Fertil. 1988 Nov;84(2):493-504. doi: 10.1530/jrf.0.0840493.
Conceptus secretory proteins (oCSP) were obtained from medium in which sheep conceptuses, collected on Day 16 of pregnancy, were cultured for 30 h. A portion of the culture medium (500 ml) was prepared for intrauterine infusion by concentrating the proteins by Amicon ultrafiltration (Mr 500 cutoff). A second portion (500 ml medium) was used to purify sheep trophoblast protein one (oTP-1). Proteins remaining after oTP-1 purification were concentrated and then passed through an anti-oTP-1 sepharose CL-4B affinity column to remove any remaining oTP-1 (oCSP-oTP-1). Serum proteins (oSP) were collected from a Day-16 pregnant ewe and diluted for infusion. Catheters were placed in the uterus of cyclic (Day 10) ewes. The following combinations of proteins were infused: 0.75 mg oCSP + 0.75 mg oSP (5 ewes), 0.75 mg oCSP - oTP-1 + 0.75 mg oSP (4 ewes), 0.05 mg oTP-1 + 1.45 mg oSP (5 ewes) and 1.5 mg oSP only (5 ewes). Infusions were twice daily on Days 12 and 13 (08:00 and 17:00 h) and once on Day 14 (08:00 h). On Day 14, ewes were injected intravenously at 08:00 h with 0.5 mg oestradiol-17 beta. Blood sampling began 30 min before oestradiol injection and continued every 30 min for 10 h. On Day 15 ewes received 10 i.u. oxytocin intravenously (08:00 h). Blood samples were collected 10 min before oxytocin and every 10 min for 1 h after oxytocin injection. Concentrations of prostaglandin (PG) F, PGE-2/PGE-1 (PGE) and 13,14-dihydro-15-keto-PGF-2 alpha (PGFM) were measured by specific radioimmunoassay. Ewes treated with oTP-1 and oCSP had longer (P less than 0.05) interoestrous intervals (27 and 25 days, respectively) compared to ewes treated with oSP and oCSP--oTP-1 (19 and 19 days, respectively) (s.e.m. = 1.56 days). These results indicate that oTP-1 alone is as potent as total conceptus secretory proteins in extending luteal maintenance. Ewes treated with oTP-1 and oCSP had no increase in PGF after oestradiol injection while production of PGF did increase 6-10 h after oestradiol in ewes treated with oSP and oCSP--oTP-1. PGFM was correlated with PGF concentrations (r = 0.57, P less than 0.01) although presence or absence of increases in production of PGFM for the treatment groups were not the same as those for PGF. No effects of treatment on PGE were detected.(ABSTRACT TRUNCATED AT 400 WORDS)
从怀孕第16天收集的绵羊孕体培养30小时的培养基中获取孕体分泌蛋白(oCSP)。通过Amicon超滤(截留分子量500)浓缩蛋白质,制备一部分培养基(500毫升)用于子宫内灌注。另一部分(500毫升培养基)用于纯化绵羊滋养层蛋白1(oTP-1)。oTP-1纯化后剩余的蛋白质被浓缩,然后通过抗oTP-1琼脂糖CL-4B亲和柱以去除任何残留的oTP-1(oCSP-oTP-1)。从怀孕第16天的母羊收集血清蛋白(oSP)并稀释用于灌注。将导管置于发情周期(第10天)母羊的子宫内。灌注以下蛋白质组合:0.75毫克oCSP + 0.75毫克oSP(5只母羊),0.75毫克oCSP - oTP-1 + 0.75毫克oSP(4只母羊),0.05毫克oTP-1 + 1.45毫克oSP(5只母羊)和仅1.5毫克oSP(5只母羊)。在第12天和第13天每天灌注两次(08:00和17:00),在第14天灌注一次(08:00)。在第14天,08:00给母羊静脉注射0.5毫克雌二醇-17β。在雌二醇注射前30分钟开始采血,并每隔30分钟持续采血10小时。在第15天,母羊静脉注射10国际单位催产素(08:00)。在催产素注射前10分钟采集血样,并在注射后每隔10分钟采集1小时。通过特异性放射免疫测定法测量前列腺素(PG)F、PGE-2/PGE-1(PGE)和13,14-二氢-15-酮-PGF-2α(PGFM)的浓度。与用oSP和oCSP - oTP-1处理的母羊(分别为19天和19天,标准误=1.56天)相比,用oTP-1和oCSP处理的母羊发情间期更长(P<0.05)(分别为27天和25天)。这些结果表明,单独的oTP-1在延长黄体维持方面与总孕体分泌蛋白一样有效。用oTP-1和oCSP处理的母羊在雌二醇注射后PGF没有增加,而在用oSP和oCSP - oTP-1处理母羊中,雌二醇注射后6 - 10小时PGF产量确实增加。PGFM与PGF浓度相关(r = 0.57,P<0.01),尽管各处理组PGFM产量增加与否与PGF不同。未检测到处理对PGE有影响。(摘要截短至400字)