BOVINE EMBRYOS PRODUCE A UROKINASE-TYPE PLASMINOGEN-ACTIVATOR

被引:16
作者
BERG, DA [1 ]
MENINO, AR [1 ]
机构
[1] OREGON STATE UNIV,DEPT ANIM SCI,CORVALLIS,OR 97331
关键词
BLASTOCYSTS; SERINE PROTEASES; TROPHOBLAST; EMBRYONIC DISKS;
D O I
10.1002/mrd.1080310104
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The type of plasminogen activator (PA) secreted by bovine embryos was identified. Day 12-14 embryos were collected from estrus-synchronized, superovulated, and naturally mated crossbred beef cows. Embryos were left intact (E) or microdissected into component embryonic discs (ED) and trophoblastic vesicles (TV). Intact embryos, ED, and TV were pre-cultured for 2 days in Minimum Essential Medium Alpha (MEM-alpha) with 10% heat-inactivated fetal calf serum, washed in serum-free MEM-alpha, and cultured individually for 5 days in 50-mu-l microdrops of MEM-alpha with 15 mg/ml bovine serum albumin. At 24 hr intervals, E, ED, and TV were observed for tissue morphology and transferred to fresh microdrops, and medium was recovered and frozen at -20-degrees-C. At the end of culture, blastocoelic fluid (BF) and embryonic tissues were recovered and frozen at -20-degrees-C. Plasminogen activator concentrations in medium, tissues, and BF were determined by using a caseinolytic assay. Antibodies to urokinase-type PA (anti-uPA) and tissue-type PA (anti-tPA), and the urokinase inhibitor, amiloride (AMR), were used to identify the type of PA produced by bovine embryonic tissues. Intact embryos and TV released more PA (P < 0.05) than ED, and tissues exhibiting expanded blastocoels released less PA (P < 0.05) than tissues with collapsed blastocoels. Blastocoelic fluid from TV exhibited more PA (P < 0.05) activity than from ED. Treatment with anti-uPA decreased PA activity (P < 0.05) in pooled medium and tissues from E compared to treatment with nonspecific immunoglobulins and anti-tPA. Amiloride completely eliminated PA activity (P < 0.05) in tissues, medium, and BF. These results suggest that day 12-14 bovine embryos secrete a urokinase-type PA.
引用
收藏
页码:14 / 19
页数:6
相关论文
共 18 条
[1]   ALTERED TROPHOBLAST FUNCTIONS IN IMPLANTATION-DEFECTIVE MOUSE EMBRYOS [J].
AXELROD, HR .
DEVELOPMENTAL BIOLOGY, 1985, 108 (01) :185-190
[2]   OVARIAN PLASMINOGEN ACTIVATOR - RELATIONSHIP TO OVULATION AND HORMONAL-REGULATION [J].
BEERS, WH ;
STRICKLAND, S ;
REICH, E .
CELL, 1975, 6 (03) :387-394
[3]   THE ANATOMY AND PHYSIOLOGY OF PRE-ATTACHMENT BOVINE EMBRYOS [J].
BETTERIDGE, KJ ;
FLECHON, JE .
THERIOGENOLOGY, 1988, 29 (01) :155-187
[4]  
BJERRUM OJ, 1975, SCAND J IMMUNOL S2, V4, P81
[5]   PLASMINOGEN ACTIVATOR SECRETION DURING MOUSE EMBRYOGENESIS [J].
BODE, VC ;
DZIADEK, MA .
DEVELOPMENTAL BIOLOGY, 1979, 73 (02) :272-289
[6]   PLASMINOGEN ACTIVATORS, TISSUE DEGRADATION, AND CANCER [J].
DANO, K ;
ANDREASEN, PA ;
GRONDAHLHANSEN, J ;
KRISTENSEN, P ;
NIELSEN, LS ;
SKRIVER, L .
ADVANCES IN CANCER RESEARCH, 1985, 44 :139-266
[7]  
DYK AR, IN PRESS J REPROD FE
[8]   RELATIONSHIP BETWEEN RELEASE OF PLASMINOGEN-ACTIVATOR AND ESTROGEN BY BLASTOCYSTS AND SECRETION OF PLASMIN INHIBITOR BY UTERINE ENDOMETRIUM IN THE PREGNANT PIG [J].
FAZLEABAS, AT ;
GEISERT, RD ;
BAZER, FW ;
ROBERTS, RM .
BIOLOGY OF REPRODUCTION, 1983, 29 (01) :225-238
[9]   PLASMINOGEN ACTIVATORS AND THEIR INHIBITORS - REGULATORS OF EXTRACELLULAR PROTEOLYSIS AND CELL-FUNCTION [J].
HART, DA ;
REHEMTULLA, A .
COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY, 1988, 90 (04) :691-708
[10]  
HEYMAN Y, 1987, MAMMALIAN PREIMPLANT, P175